Understanding the Role of Pyruvate Dehydrogenase Kinase 3 in Gynecological Cancers

Understanding the Role of Pyruvate Dehydrogenase Kinase 3 in Gynecological Cancers | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Understanding the Role of Pyruvate Dehydrogenase Kinase 3 in Gynecological Cancers Medha Bairy, Aishath Shaheeda, Padmanaban S. Suresh, Roshan Mascarenhas, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6429535/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Pyruvate dehydrogenase kinase 3 (PDK3) is a key regulatory enzyme of the pyruvate dehydrogenase complex that determines the fate of a cell towards oxidative phosphorylation or aerobic glycolysis through metabolic reprogramming. This study was designed to examine the role of PDK3 in gynecological cancers via in silico analysis and its conformation via in vitro experiments. Analysis of gynecological cancer datasets from the TCGA and CCLE databases suggested significant upregulation of PDK3 in breast, cervical, and endometrial cancer samples compared with the corresponding normal samples. KEGG pathway enrichment analysis revealed that pathways associated with cancers, viral carcinogenesis, and human papilloma virus infection were highly enriched. PDK3 upregulation affected the overall survival of breast and cervical cancer patients according to Kaplan–Meier survival analysis. PDK3 upregulation affected the proportion and distribution of immune cells in breast, cervical, and endometrial cancer. Furthermore, we experimentally showed that PDK3 knockdown inhibits the proliferation and growth of SiHa cells. Overall, we found that PDK3 may be an oncogene, and measuring its expression may be of prognostic and therapeutic significance in gynecological cancers. PDK3 gynecological cancer metabolic reprogramming oncogene proliferation growth Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction The incidence of gynecological cancer (GC), a cancer affecting multiple organs of the female reproductive system, has substantially increased, contributing to 15.25% of all cancer cases reported and 15.77% of cancer-related deaths among women worldwide [ 1 ]. The global burden of gynecological cancer is predicted to increase by 49.61%, and the associated mortality is proposed to increase to 77.23% by the year 2050 [ 2 ]. Among all gynecological cancers, breast cancer continues to be the primary cause of cancer-associated death, followed by cervical cancer, ovarian cancer and endometrial cancer [ 1 ]. Metabolic reprogramming is an important hallmark of cancer, supporting the energy demand of rapidly proliferating and growing cancer cells. One such metabolic rewiring is the sustenance of a higher rate of glycolysis despite having sufficient oxygen, termed the Warburg effect [ 3 ]. Cancer cells have been shown to display rewired metabolic pathways, perhaps shifting away from oxidative phosphorylation and toward glycolysis. The pyruvate dehydrogenase kinase (PDK) family of proteins regulates the pyruvate dehydrogenase complex (PDHC), a critical regulatory enzyme that links glycolysis with the citric acid cycle. By controlling this complex, PDK plays a pivotal role in shifting cellular respiration toward the Warburg effect [ 4 ]. The PDK family consists of four isoforms, namely, PDK1, PDK2, PDK3, and PDK4, which share 70% homology and exhibit differential tissue expression, all of which are present in the mitochondrial matrix [ 5 ]. PDK inactivates PDHC by phosphorylating serine residues at three different sites (Ser293, Ser300, and Ser232) in its E1α subunit [ 6 ]. The inactivation of PDHC results in altered metabolic pathways, which may shift away from oxidative phosphorylation and toward glycolysis, supporting the energy demand of rapidly proliferating and growing cancer cells [ 7 ]. The activity of PDK is proportional to its binding efficiency to the lipoyl domain of the E2 subunit in PDH. Among all the isoforms, pyruvate dehydrogenase kinase 3 (PDK3) has the highest affinity and is not affected by the presence of the substrate pyruvate [ 8 ]. Recent findings indicate a multifaceted role of PDK3 in cancer progression [ 6 ]. PDK3 is upregulated in cancers such as colon [ 9 ], acute myeloid leukemia [ 4 ], gastric [ 10 ] and urothelial carcinomas [ 11 ]. PDK3 promotes tumorigenesis by stimulating cell proliferation, angiogenesis, invasion, and glycolysis in multiple cancer types [ 12 ]. Immunohistochemical staining has shown a direct correlation between PDK3 expression and disease severity in colon cancer cells [ 9 ]. PDK3 overexpression reportedly contributes to chemoresistance and cancer recurrence through the combined action of Hypoxia-Inducing Factor α ( HIF-α ) and Heat Shock Factor 1( HSF1 ) [ 8 , 13 ]. PDK3 also regulates the PI3K-Akt pathway to promote tumorigenic characteristics [ 14 ]. PDK3 inhibits the T-cell-mediated immune response and suppresses the expression of Programmed Death Ligand-2 (PDL-2) by inhibiting the Jak1/STAT 1 pathway, thereby promoting cancer growth [ 15 ]. PDK3 could be used as a potential target in cancer because of its multifaceted tumor-promoting functions. Nevertheless, the significance of PDK3 in gynecological cancers remains poorly understood. To address this gap, we carried out a comprehensive analysis to understand the role of PDK3 in gynecological cancers. We systematically studied the expression, regulation, clinical utility, interaction networks, biological functions, immune infiltration, and chemical activity of PDK3 to target PDK3 in gynecological cancers. Finally, we performed shRNA-mediated PDK3 knockdown studies to assess the role of PDK3 in cervical cancer. Materials and methods Data acquisition and processing PDK3 expression data were sourced from the Genotype-Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA) databases [ 16 ]. The GTEx database offers a detailed catalogue of gene expression levels among various tissue types, facilitating the comparison of PDK3 expression status in different tissue types. Box plots showing the differential expression of PDK3 between normal and tumor samples were generated via the TIMER2.0 database under the ‘Gene DE’ module [ 17 ]. The genomic, transcriptomic, and clinical data from TCGA were retrieved via UCSC Xena and GEPIA [ 18 ]. PDK3 expression between tumors and normal tissues was tested for significance via the Wilcoxon rank-sum test. Statistical significance was set at a p value of < 0.05. The SRPlot tool was used to represent the obtained result in box plot form [ 19 ]. Furthermore, PDK3 expression data were cross-validated with datasets from the Cancer Cell Line Encyclopedia (CCLE) [ 20 ]. Exploration of genetic and epigenetic alterations The IntOGen [ 21 ] database was analysed to retrieve mutations connected with PDK3 in the pancancer samples. The TCGA database was used to visualize mutations in different cancer types. The TIMER 2.0 platform was used to visualize the frequency and types of mutations. Additionally, copy number variations (CNVs), amplifications, and deletions linked to PDK3 were examined via the GSCA database [ 22 ]. We next analysed the association between DNA methylation and PDK3 expression. PDK3 promoter DNA methylation in normal and tumor samples was tested via the UALCAN database [ 23 ]. We specifically selected CpG sites located within 1000 base pairs upstream of the PDK3 promoter. The correlation between methylation and expression was tested via Spearman correlation analysis [ 24 ]. To explore the post-transcriptional regulation of PDK3 , we performed an extensive search of miRNAs that target PDK3 via three miRNA target scanning tools, namely, miRNet [ 25 ], miRDB [ 26 ], and TargetScan [ 27 ]. The common miRNAs found in these three databases were identified via the IteractiVenn tool [ 28 ]. Coexpression and protein‒protein interaction network construction We used the GeneMANIA [ 29 ] and BioGRID databases [ 30 ] to construct the PDK3 and its coexpressed gene network. The STRING web server was used to construct the protein‒protein interaction network among coexpressed genes [ 31 ]. Functional linkages and interactions involving PDK3 were interpreted by combining the interaction data with correlation coefficients from the TIMER2.0 database, which focused on gynecological cancer types. The Cancerhallmarks [ 32 ] tool was used to visualize the enriched cancer hallmarks related to PDK3 coexpressed genes. This method allows for a thorough examination of PDK3 's regulatory network and possible contribution to the development of cancer. Transcriptional regulator identification and functional enrichment analysis TFlink [ 33 ] was used to identify the transcription factors regulating PDK3 . By combining information from chromatin immunoprecipitation (ChIP) investigations with computational predictions, TFlink offers information on putative transcription factors that may control genes of interest. The biological pathways linked to these regulators and the enrichment of gene ontology (GO) terms were evaluated using ShinyGO [ 34 ]. Clinical implications and survival outcomes The correlation between PDK3 expression and cancer progression stage was determined via the GDC-TCGA datasets accessible via GEPIA [ 35 ]. The prognostic significance of PDK3 expression was evaluated via Kaplan‒Meier plotter for survival analysis [ 36 ]. Immune infiltration and drug interaction analysis The TISIDB tumor-immune system interaction database was used to analyse immune cell infiltration patterns and PDK3 expression [ 37 ]. The relationships between PDK3 expression and 24 distinct tumor-infiltrating lymphocyte subsets were examined via the TIMER 2.0 program. SRplot was used to create heatmaps from the data. Finally, the GSCA database was used to examine the relationship between PDK3 expression and drug response [ 22 ]. Data on drug‒gene correlations from the Cancer Therapeutics Response Portal (CTRP) and Genomics of Drug Sensitivity in Cancer (GDSC) databases are integrated by the GSCA. SRplot was used to create a bubble plot that represented the data. The various databases and their links used in the present study are listed in Table 3 . Cell culture and transfection SiHa cells were cultured and maintained according to ATCC guidelines. To generate the PDK3 knockdown cell line, SiHa cells (2×10 6 ) were seeded in a 6-well plate and incubated at 37°C in a 5% CO 2 environment. After 24 hours, the cells were transfected with a shRNA plasmid against PDK3 (sc-39029-SH; Santa Cruz). Control plasmid (sc-108060; Santa Cruz)-transfected cells served as controls. We used Lipofectamine 3000 (Thermo Fisher Scientific, USA) to carry out the transfections according to the manufacturer's protocol. PDK3 -knockdown clones were isolated by culturing the cells in 2.0 µg/ml puromycin for 14 days [ 38 ]. Confirmation of PDK3 knockdown Total RNA was isolated using TRIzol reagent (Thermo Fisher Scientific, USA). PDK3 Hs00178440_m1; Invitrogen) knockdown was confirmed by TaqMan-based qRT-PCR using 25ng of RNA with β-actin as an endogenous control. Gene expression was calculated using the ΔΔCt method, where ΔCt = Ct (target gene) - Ct (reference gene), ΔΔCt = ΔCt test - ΔCt control. The fold change (FC) was calculated as FC = 2^(-ΔΔCt) [ 39 ]. After being washed in ice-cold PBS, PDK3 knockdown and vector-transfected cells were lysed in RIPA buffer containing a protease inhibitor. Using SDS‒PAGE, approximately 40 µg of protein was separated and then blotted onto a nitrocellulose membrane. The membranes were incubated overnight at 4°C in the presence of primary antibodies against PDK3 (1:2000 dilution, ABclonal, USA) and anti-ACTB (1:5000 dilution, ABclonal, USA). The membranes were then incubated for two hours at room temperature with an HRP-conjugated secondary antibody after being washed with TBST. To visualize the PDK3 protein, Clarity Western ECL Substrate (Bio-Rad, USA) was utilized. An iBright imaging system (Invitrogen, USA) was used to capture images [ 39 ]. Proliferation assay Trypsinization and trypan blue staining were performed on the cells every 24 hours, and a Neubauer chamber was used to count living cells under a microscope. Omni calculator was used to determine the doubling times of the control and knockdown cells [ 40 ]. Colony formation assay Approximately 500 cells were cultivated in a 6 cm cell culture plate for two weeks in full media. Following the disposal of the culture media and washing with PBS, the colonies were stained for ten to fifteen minutes with 0.5% crystal violet in methanol staining solution. After extra stain was removed and the samples were washed with PBS, the number of stained colonies was calculated [ 38 ]. Results PDK3 is overexpressed in gynecological cancers Using the GTEX database, we first examined the expression of PDK3 in several tissue types. PDK3 was expressed at varying levels in various human tissues (Fig. 1 A). Pancancer analysis of PDK3 expression via the TIMER 2.0 tool revealed that PDK3 was upregulated in tumor samples compared with normal samples in the TCGA cohort (Fig. 1 B). We found that PDK3 mRNA levels were significantly elevated in uterine corpus endometrial carcinoma (TCGA-UCEC), cervical squamous cell carcinoma (TCGA-CESC), and invasive breast carcinoma (TCGA-BRCA) (Fig. 1 C). Interestingly, PDK3 expression did not significantly change in ovarian cancer (Fig. 1 D). High expression of PDK3 was observed in different gynecological cell lines (Fig. 1 E). Regulation of PDK3 across gynecological cancers Mutation analysis via IntOGen identified 129 mutations in PDK3 , with missense mutations being the most frequent (60%), followed by synonymous (29%), truncating (8%), and splice-site (3%) mutations. Hotspot mutations were detected, especially in the functional domains of PDK3 , indicating that these mutations might affect the function of PDK3 (Fig. 2 A). According to TCGA dataset analysis, mutations in PDK3 were absent in breast cancer patients, 0.68% in cervical cancer patients, and slightly more frequently (6.4%) in endometrial cancer patients (Fig. 2 B). The low mutation prevalence indicates that although genetic modifications do occur, the total mutation rate across tumors is still low, suggesting that PDK3 dysregulation in the TCGA-CESC (0.68%) and TCGA-BRCA (0%) cohorts might originate from other mechanisms, such as DNA methylation or miRNA (Fig. 2 C). To investigate factors other than mutations that potentially regulate the expression of PDK3 , we examined copy number variations (CNVs) in endometrial, cervical, and breast malignancies. Remarkably, a sizable portion of patients presented gains at the PDK3 locus; amplifications were detected in almost 17% of patients with breast cancer, 16% of patients with cervical cancer, and approximately 14% of patients with endometrial cancer (Fig. 2 D) (Table 1 ). Linking these CNVs with PDK3 expression revealed that there was a positive correlation between these CNVs and PDK3 expression, especially in endometrial cancer (Spearman R coefficient 0.19, p value < 0.05) and invasive breast carcinoma (Spearman R coefficient 0.09, p value < 1001) (Fig. 2 E). Cervical cancer had a Spearman R coefficient of 0.02, suggesting that the upregulation of PDK3 in cervical cancer may be independent of CNV. These data indicate that CNVs may significantly contribute to the overexpression of PDK3 in breast cancer. Table 1 The GSCA database showing various percentages and types of SNVs and CNVs at the PDK3 locus in different gynaecological cancers. Percentages and types of SNVs and CNVs at the PDK3 locus in different gynecological cancers SNVs Cancer type Effective mutations Non effective mutations Sample size Percentage BRCA 0 3 1026 0 CESC 2 1 291 0.687285223 UCEC 36 12 531 6.779661017 CNVs Cancer type Total amplification (%) Total Deletion (%) Heterozygous amplification (%) Heterozygous deletion (%) Homozygous amplification (%) Homozygous deletion (%) BRCA 17.2222222 17.5 15.6481481 17.1296296 1.5740741 0.37037 CESC 16.6101695 21.6949153 14.2372881 21.6949153 2.3728814 0 UCEC 14.1001855 15.7699443 14.1001855 15.3988868 0 0.371058 Additionally, we examined the effects of epigenetic changes by evaluating DNA methylation patterns at the PDK3 promoter in several gynecological malignancies (Supplementary Fig. 1A). Our analysis revealed significant methylation at most of the CpG sites (p value < 0.05) in breast cancer and uterine cancer (Supplementary Fig. 1B) (Table 2 ). Promoter methylation analysis revealed that the promoter was not hypermethylated despite a significant increase in methylation (a beta value cut-off of 0.7–0.5 indicated hypermethylation, and 0.3–0.25 indicated hypomethylation), suggesting that additional epigenetic elements, such as miRNAs, are involved in regulating gene expression (Supplementary Fig. 1C). Correlation analysis revealed that PDK3 expression and promoter methylation were strongly negatively correlated in the TCGA-BRCA cohort and TCGA-CESC cohort (Supplementary Fig. 1D). These findings suggest that different epigenetic factors play a role in controlling the expression of PDK3 in individual cancer types. Table 2 The OncoDB database showing methylation levels of PDK3 observed on CpGs in different cancers. (Probes in red colour shows the significant differential methylation). Probe Position Average of Cancer Sample Average of Normal Sample p-value BRCA cg20258741 24464729 0.41 0.44 4.7e-06 cg23143524 24464960 0.55 0.54 2.6e-01 cg02539591 24464975 0.41 0.42 2.0e-01 cg02623256 24465114 0.39 0.39 8.3e-01 cg06721573 24465117 0.40 0.44 6.2e-04 cg01212677 24465173 0.46 0.50 2.1e-03 cg15308487 24465175 0.31 0.33 1.8e-02 cg15345580 24465177 0.23 0.20 3.0e-04 cg22969661 24465359 0.51 0.51 9.8e-01 cg00114625 24465361 0.55 0.56 8.0e-03 cg18414950 24465779 0.48 0.58 2.0e-12 cg25863147 24466244 0.47 0.57 1.6e-11 cg13777527 24539503 0.87 0.94 1.6e-12 CESC cg20258741 24464729 0.45 0.38 2.2e-01 cg23143524 24464960 0.58 0.47 1.3e-01 cg02539591 24464975 0.44 0.37 3.4e-01 cg02623256 24465114 0.43 0.38 3.5e-01 cg06721573 24465117 0.41 0.47 5.0e-07 cg01212677 24465173 0.48 0.44 3.3e-01 cg15308487 24465175 0.33 0.30 4.2e-01 cg15345580 24465177 0.28 0.14 1.0e-03 cg22969661 24465359 0.53 0.43 9.1e-03 cg00114625 24465361 0.55 0.51 2.1e-09 cg18414950 24465779 0.53 0.51 8.4e-01 cg25863147 24466244 0.61 0.67 4.6e-01 cg13777527 24539503 0.89 0.94 3.6e-01 UCEC cg20258741 24464729 0.43 0.42 2.5e-01 cg23143524 24464960 0.57 0.53 8.6e-04 cg02539591 24464975 0.41 0.41 9.8e-01 cg02623256 24465114 0.41 0.35 3.2e-07 cg06721573 24465117 0.39 0.42 3.9e-03 cg01212677 24465173 0.48 0.51 3.9e-02 cg15308487 24465175 0.33 0.36 1.7e-02 cg15345580 24465177 0.25 0.21 6.5e-03 cg22969661 24465359 0.53 0.50 2.2e-03 cg00114625 24465361 0.56 0.54 2.1e-02 cg18414950 24465779 0.51 0.55 1.7e-05 cg25863147 24466244 0.69 0.68 6.3e-01 cg13777527 24539503 0.87 0.93 1.6e-12 Table 3 Table showing all the databases and tools used in the current study. Databases/Sites used in the study Links Genotype-Tissue Expression (GTEx) https://gtexportal.org/home/ TIMER2.0 http://timer.cistrome.org/ UCSC Xena https://xena.ucsc.edu/ UALCAN https://ualcan.path.uab.edu/ Cancer Cell Line Encyclopedia (CCLE) https://sites.broadinstitute.org/ccle/ OncoDB https://oncodb.org/ Gene Set Cancer Analysis (GSCA) https://guolab.wchscu.cn/GSCA/#/ GeneMANIA https://genemania.org/ BioGRID https://thebiogrid.org/ STRING https://string-db.org/ Cancer hallmarks https://cancerhallmarks.com/ TFlink https://tflink.net/ GEPIA http://gepia.cancer-pku.cn/ Kaplan-Meier plotter https://kmplot.com/analysis/ TISIDB http://cis.hku.hk/TISIDB/ IntOGen https://www.intogen.org/search SRplot https://www.bioinformatics.com.cn/srplot Venny 2.1 https://bioinfogp.cnb.csic.es/tools/venny/ ShinyGO 0.81 https://bioinformatics.sdstate.edu/go/ miRNET https://www.mirnet.ca/upload/MirUploadView.xhtml miRDB https://mirdb.org/ TargetScan https://www.targetscan.org/vert_80/ Cell doubling time calculator https://www.omnicalculator.com/ We investigated the role of microRNAs (miRNAs) as potential regulators of PDK3 expression. Using data from the miRDB, miRNET and TargetScan databases, we identified 11 miRNAs that could target PDK3 (Fig. 3 A) with different target scores (a score of 70–100 indicating high confidence) (Fig. 3 B). Among the 11 miRNAs, hsa-miR-525 and hsa-miR-369 were downregulated in all three cancers, and hsa-miR-186 and hsa-miR-494 were downregulated only in TCGA-BRCA and TCGA-CESC, respectively. hsa-miR-205 and hsa-miR-194 were downregulated only in the TCGA-BRCA cohort and TCGA-CESC cohort, respectively (Fig. 3 C). Hence, downregulation of these miRNAs might reduce their suppressive effect on the PDK3 gene. Co-expression and interaction analysis of PDK3 Using GeneMANIA (Fig. 4 A) and BioGRID (Fig. 4 B), we constructed a coexpression network of PDK3 . The STRING tool was used to generate the PPIN (Fig. 4 C). Protein‒protein interaction (PPI) network analysis revealed a complex network consisting of 14 nodes and 34 edges. The network demonstrated significant PPI enrichment, indicating nonrandom interactions among the identified proteins. Furthermore, the protein‒protein interaction network (PPIN) constructed from the overlapping genes revealed a direct interaction between PDK3 and 8 members (Fig. 4 D). Thus, our in silico analysis revealed strong interactions of PDK3 with DLAT, DLD, PDHA1, PDHB, PDHX, PDK1, PDK2, and PDK4. This finding was supported by correlation analysis, which revealed positive correlations between PDK3 and DLAT, DLD, PDHA1, PDHB, PDHX, and PDK1 (Fig. 4 E). Furthermore, a cancer hallmark enrichment plot of PDK3 coexpressed genes revealed significant involvement in reprogramming and energy metabolism (Fig. 4 F). PDK3 modulates critical signalling pathways in cancer progression In silico analysis revealed PDK3 as part of many key oncogenic signalling pathways. We used ChIP-seq data available at TFlink to identify key transcription factors (TFs) regulating PDK3 expression. We determined that PDK3 possesses binding sites close to 414 transcription factors (Supplementary Table 1). The upstream regulators of PDK3 are predominantly connected with pathways in cancer, the cell cycle, Wnt signalling, miRNAs in cancer, breast cancer, and HPV infection, as shown by KEGG pathway enrichment analysis (Supplementary Fig. 2A). Gene Ontology (GO) term analysis further suggested that enrichment of RNA biosynthesis, differential transcription, and macromolecule biosynthesis were the top hit BPs (Supplementary Fig. 2B). Significant enrichment in transcription coregulator activity and different transcription factor binding activities was observed for PDK3 targets at the molecular function (MF) level. The majority of these functions were linked to cellular components that regulate DNA epigenetically (Supplementary Fig. 2C & 2D). Clinical significance of PDK3 across gynecological cancers The clinical significance of aberrant PDK3 expression in gynecological cancers was evaluated via the GEPIA tool. Correlation analysis of PDK3 expression with overall survival revealed significant prognostic implications in breast and cervical cancer patients. Notably, increased PDK3 expression in the TCGA-CESC cohort and decreased PDK3 expression in the TCGA-BRCA and TCGA-UCEC cohorts correlated with improved survival outcomes (p < 0.05) (Supplementary Fig. 3A). We next tested the association of PDK3 expression with cancer stage in breast, cervical, and endometrial cancer. There were no significant differences between PDK3 expression and cancer stage (F value < 2.5) (Supplementary Fig. 3B). PDK3 expression can affect the abundance of immune cells We generated a violin plot via TISIDB to evaluate PDK3 expression in different immune subtypes, classifying immune subtypes into six classes: wound healing (C1), IFN-gamma dominant (C2), inflammatory (C3), lymphocyte depleted (C4), immunologically quiet (C5), and TGF-β dominant (C6). PDK3 expression correlated with five immunological subtypes (C1, C2, C3, C4 and C6) in the TCGA-BRCA cohort and three immunological subtypes (C1, C2, and C4) in the TCGA-CESC cohort, indicating that PDK3 expression differs according to the subtype of the immune microenvironment (Fig. 5 A). TIMER 2.0 database analysis revealed an inverse relationship between PDK3 expression and the numbers of NK cells and CD8 + T cells in the TCGA-BRCA cohort. In the TCGA-CESC cohort, unpolarized memory B cells were positively correlated with PDK3 expression. Additionally, PDK3 expression is positively correlated with antitumour M1 macrophages and negatively correlated with protomeric M2 macrophages. In addition, PDK3 overexpression was significantly negatively correlated with the expression of memory B cells and NK cells. In the TCGA-UCEC cohort, a significant negative correlation between NK cells and PDK3 was observed (Fig. 5 B). PDK3 -drug interactions Higher PDK3 expression was positively correlated with the expression of most GDSC drugs and one of the CTRP drugs, dasatinib, indicating that increased expression of this gene may lead to resistance to these drugs. A negative correlation of PDK3 was observed with most of the CTRP drugs and two of the GDSC drugs, GSK690693 and PLX470, suggesting that increased expression of this gene might lead to drug sensitivity (Fig. 5 C&D). PDK3 knockdown in SiHa cells We previously reported that PDK3 is significantly overexpressed in cervical cancer patient samples (Srinath et al., 2024). This finding was further confirmed by in silico differential expression analysis of the TCGA-CESC datasets. qRT‒PCR and western blot analysis revealed significant expression of PDK3 in SiHa cells. Hence, we knocked down PDK3 in SiHa cells to investigate its biological properties. PDK3 knockdown in SiHa cells was confirmed by qRT‒PCR, which revealed an 85% decrease in the expression of PDK3 (Fig. 6 A). The knockdown effect was then confirmed by western blot analysis (Fig. 6 B & C). PDK3 knockdown inhibits the growth and proliferation of SiHa cells PDK3 knockdown in SiHa cells was confirmed by qRT‒PCR and western blotting (Fig. 6 A‒6C). The impact of PDK3 on proliferation and growth was quantified via growth curve analysis and colony formation assays. The growth curve analysis revealed a significant reduction in cell number upon PDK3 knockdown in SiHa cells. In contrast, the scrambled control-treated cells exhibited significant cell proliferation (Fig. 6 D). The cell doubling time in scrambled and PDK3 -knockdown cells was 39.27 hrs. and 173.45 hrs., respectively. We next investigated the impact of PDK3 knockdown on the growth of SiHa cells via a colony formation assay. We found that, compared with scrambled treatment, PDK3 knockdown significantly reduced the number of colonies (Fig. 6 E and F). Discussion The involvement of PDK3 in cancer progression has gained significant attention because of its critical role in regulating cellular metabolism, especially in inhibiting PDHC. In the present study, we investigated the regulation, functional implications, and clinical significance of aberrant PDK3 expression in gynecological cancers through in silico analysis and in vitro experiments. According to earlier research, PDK3 is overexpressed in the stomach [ 10 ], colon [ 9 ], lung [ 41 ], cervix [ 42 ], breast [ 43 ], and urinary bladder [ 11 ] and may contribute to metabolic reprogramming and tumor progression [ 13 ]. Despite its aberrant expression in multiple cancers [ 8 , 13 , 15 ], its role in gynecological cancers has not been well investigated. Our in silico analysis of the TCGA-BRCA, TCGA-UCEC, and TCGA-CESC datasets revealed notable overexpression of PDK3 in cancer samples compared with normal samples. The Cell Line Encyclopedia (CCLE) also confirmed relatively high PDK3 expression in gynecological cancer cell lines, further emphasizing the oncogenic potential of PDK3 . Genetic and epigenetic analyses of PDK3 revealed that somatic PDK3 mutations were relatively uncommon in all of the cancer samples examined. Considerable CNV gains were observed in the TCGA-CESC, TCGA-UCEC, and TCGA-BRCA samples. Spearman correlation analysis revealed a positive correlation of PDK3 expression only in the TCGA-UCEC cohort and TCGA-BRCA cohort. Hence, CNV gain may have contributed to PDK3 overexpression in the TCGA-UCEC and TCGA-BRCA cohorts but not in the TCGA-CESC cohort. This finding provides the scope for further investigation of the cause of PDK3 overexpression in the TCGA-CESC cohort. Aberrant DNA methylation has been shown to influence gene expression, with hypermethylation contributing to gene silencing and hypomethylation contributing to gene upregulation [ 40 , 44 , 45 ]. Although our DNA methylation studies revealed significantly increased methylation in the normal tissues of all three cancers at specific CpG sites, the findings were inconsistent across all the CpG sites. The beta value cut-off indicated that neither significant hypermethylation nor hypomethylation was present in the tumor samples. However, according to the Spearman correlation analysis, the TCGA-BRCA and TCGA-CESC cohorts presented significant negative correlations with promoter methylation and PDK3 expression. We previously reported that PDK3 is an upregulated gene in cervical cancer [ 42 ]. More detailed studies are needed to understand the relationship between promoter DNA methylation and PDK3 expression. MicroRNA interaction and regulatory network analyses have identified multiple microRNAs that possibly target PDK3 expression in gynecological cancers. Importantly, hsa-miR-186-5p, hsa-miR-194-5p, hsa-miR-7-5p and hsa-miR-494-3p have been shown to target PDK3 . hsa-miR-186-5p downregulates PDK3 expression in glioma [ 46 ], whereas hsa-7-5p has been studied for its role in suppressing PDK3 expression in TCGA-HNSC [ 47 ]. Our previous research revealed that hsa-miR-494-3p targets PDK3 in cervical cancer and that the binding of PDK3 by hsa-miR-494-3p results in the reversal of tumorigenic properties through metabolic reprogramming [ 42 ]. This differential miRNA regulation suggests that the diverse miRNAs act to tightly regulate PDK3 expression among different cancer types to accommodate distinct metabolic adaptations. Coexpression analysis of PDK3 highlighted the potential interaction of PDK3 with PDK1, PDK2, PDK4, PDHX, PDHB, PDHA1, DLAT, and DLD. The interaction of PDK3 with genes such as PDHX, PDHB, DLAT and DLD may be important for promoting metabolic reprogramming and requires detailed investigation. The hallmark enrichment analysis highlighted the role of PDK in metabolic reprogramming in cancer cells. Hence, more thorough mechanistic research is needed to understand how PDK3 and its coexpressed genes contribute to metabolic reprogramming and the development of cancer. Prognostic markers are clinical indicators used in oncology to determine a patient's risk of disease recurrence following initial therapy. Previous studies have shown that PDK3 influences patient survival in urothelial carcinoma [ 11 ], colon cancer [ 9 ], and cholangiocarcinoma [ 48 ]. Thus, we explored the prognostic significance of PDK3 in breast, cervical, and endometrial cancer using the TCGA dataset. In TCGA-UCEC patients, the overexpression of PDK3 indicated a poor survival outcome. However, PDK3 overexpression contributed to better overall survival in the TCGA-TCGA-BRCA and TCGA-TCGA-CESC datasets. Immune cells and the tumor microenvironment are decisive in determining tumor progression and metastasis [ 49 ]. Tumor-infiltrating immune cells have been reported to influence patient survival. Tumor-associated macrophages and neutrophils exhibit either protumorigenic (M2 and N2) or antitumorigenic (M1 and N1) effects on the basis of their polarization status [ 50 , 51 ]. Natural killer (NK) cells cause potent cytolysis and antitumour activity in cancer cells [ 52 ]. Similarly, CD8 + and CD4 + T cells exhibit increased antitumour activity through granular exocytosis [ 53 ]. We analysed the link between PDK3 expression and immune cell abundance via TCGA datasets. In the TCGA-BRCA cohort, significantly lower numbers of NK cells and CD8 + T cells were observed, which might be responsible for the poor survival outcome. Similarly, in TCGA-UCEC patients, the reduced survival can also be attributed to the significant downregulation of CD8 + T cells. The TCGA-CESC dataset revealed significantly increased numbers of unpolarized macrophages and significant downregulation of NK cells. These preliminary observations on tumor infiltration and its involvement in tumor prognosis upon PDK3 expression need to be explored in detail. Previous studies reported the role of PDK3 in therapy resistance, such as in colon cancer [ 9 , 15 ], gastric cancer [ 13 ], and melanoma [ 54 ]. Many drugs, such as doxorubicin, etoposide, and COL-3, are negatively correlated with PDK3 expression. More detailed studies are needed to understand the role of PDK3 in the development of therapeutic resistance to these drugs. Finally, we investigated the biological functions of PDK3 in SiHa cells. The knockdown of PDK3 in SiHa cells inhibited both the proliferation and growth of SiHa cells. Our study revealed that PDK3 may promote tumor growth by increasing cell proliferation by decreasing the cell doubling time. Our findings are consistent with previous reports in other cancers, such as urothelial carcinoma [ 11 ] and colon cancer [ 9 ]. Overall, we report a procarcinogenic function of PDK3 in gynecological cancer. Conclusion Our study investigated the role of PDK3 in gynecological cancers through extensive computational analysis followed by experimental validation. The gene expression data from multiple databases consistently revealed its overexpression, which emphasizes its role as a potential oncogene. Furthermore, genomic and epigenomic changes have been shown to influence PDK3 expression. PDK3 expression may influence immune cell abundance and clinical outcomes in breast, uterine, endometrial and cervical cancers. Our study identified potential drugs that can be used to treat cancers that overexpress PDK3 . The results of the computational analysis followed by experimental validation suggest an oncogenic role for PDK3 in cervical cancer. These findings clarify the role of PDK3 as a promising biomarker in gynecological cancers. Declarations Funding This work was supported by the Wellcome Trust DBT India Alliance, Government of India (Grant no: IA/I/22/1/506240) , awarded to Dr. Shama Prasada K, and support from Joint CSIR-UGC NET Junior Research Fellowship, Government of India (Reference ID 221610058423), and Dr. TMA Pai structured Ph.D. fellowship. Competing interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Author contributions Dr. Shama Prasada Kabekkodu: Conceptualization, Project administration, Resources, Supervision, Visualization, Writing – review and editing. Medha Bairy: Formal analysis, Investigation, Software, Validation, Writing – original draft, Visualization. Aishath Shaheeda: Formal analysis, Investigation, Software, Validation, Writing – original draft, Visualization. Padmanaban S. Suresh: Writing – review and editing. Roshan Mascarenhas: Writing – review and editing. Ethics approval The authors declare that they have no ethical statements to disclose since the research does not include any animal or human studies. Data availability statement The authors declare that all other data supporting the findings of this study are available within the article and its supplementary information files. References H. Sung, J. Ferlay, R.L. Siegel, M. Laversanne, I. Soerjomataram, A. Jemal, F. 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A Differential methylation of the PDK3 promoter visualized using UALCAN B Heatmap of PDK3 methylation at different CpG sites in all 3 cancers analysed using TCGA datasets from the UCSC Xena browser and plotted using an SR plot. C Differential methylation levels of PDK3 observed in different cancers on considering CpGs within 1 kb upstream of the PDK3 promoter. D Spearman's correlation between PDK3 expression and promoter methylation was analysed using TCGA datasets from the UCSC Xena browser and plotted using an SR plot. SuplementaryFig.2.tif Supplementary Fig. 2 Functional enrichment analysis of PDK3 upstream transcriptional regulators. A KEGG pathway analysis. B Biological processes. C Molecular processes. D Cellular components. SuplementaryFig.3.tif Supplementary Fig. 3 Overall survival and stagewise expression of PDK3 . An Overall survival analysis of PDK3 based on gene expression in different cancers plotted using KM plotter. B PDK3 gene expression in different clinical stages of cancer visualized using GEPIA database. SuplementaryTable.1.docx Supplementary Table 1 Transcription factors that interactwith PDK3 were obtained from the TFlink database. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6429535","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":456329352,"identity":"44494aaa-af6c-49c4-85ff-36c2b583692a","order_by":0,"name":"Medha Bairy","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Medha","middleName":"","lastName":"Bairy","suffix":""},{"id":456329353,"identity":"006e8c31-5084-48ed-9a9a-579775dbc110","order_by":1,"name":"Aishath Shaheeda","email":"","orcid":"","institution":"Manipal Academy of Higher Education","correspondingAuthor":false,"prefix":"","firstName":"Aishath","middleName":"","lastName":"Shaheeda","suffix":""},{"id":456329354,"identity":"308fa772-3fdb-4f81-9d22-6428184b78c3","order_by":2,"name":"Padmanaban S. 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A \u003c/strong\u003e\u003cem\u003ePDK3\u003c/em\u003eexpression across different human tissues retrieved from the GTEx dataset of the HPA database. \u003cstrong\u003eB \u003c/strong\u003e\u003cem\u003ePDK3\u003c/em\u003e expression across different human tissues retrieved from the TIMER 2.0 database. \u003cstrong\u003eC \u003c/strong\u003e\u003cem\u003ePDK3\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003egene expression across different gynecologicalcancers retrieved from the UCSC Xena browser: Increased expression of \u003cem\u003ePDK3\u003c/em\u003ein BRCA, CESC and UCEC was observed in tumors compared with normal tissues. \u003cstrong\u003eD\u003c/strong\u003e Cross-validationof \u003cem\u003ePDK3\u003c/em\u003e expression in OV via the GEPIA database. \u003cstrong\u003eE \u003c/strong\u003e\u003cem\u003ePDK3\u003c/em\u003e expression in gynecologicalcancer cell lines retrieved from the CCLE database.\u003c/p\u003e","description":"","filename":"Fig.1.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/68363162a318b47e78ba7a23.jpg"},{"id":82897312,"identity":"b512e87f-c547-469c-9b1c-9cbb847c1c35","added_by":"auto","created_at":"2025-05-16 13:03:42","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":182648,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTypes of genetic variation observed at the \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003elocus: \u003c/strong\u003eOverall mutations in \u003cem\u003ePDK3\u003c/em\u003e visualized via theIntOGen database.\u003cstrong\u003e B \u003c/strong\u003eGenetic variation analysis of the \u003cem\u003ePDK3\u003c/em\u003egene with the percentage of \u003cem\u003ePDK3\u003c/em\u003e mutations visualized via the TIMER 2.0 database. \u003cstrong\u003eC \u0026amp; D \u003c/strong\u003ePercentage and types of SNVs and CNVs observed at the \u003cem\u003ePDK3\u003c/em\u003elocus in gynecologicalcancer types visualized via the GSCA database. \u003cstrong\u003eE \u003c/strong\u003eCorrelations of CNV with \u003cem\u003ePDK3\u003c/em\u003e expression in different cancers visualized through the GSCA database.\u003c/p\u003e","description":"","filename":"Fig.2.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/e849f35c16f10d66e77b6546.jpg"},{"id":82899506,"identity":"2579faf7-0ee3-4025-861b-b6d8911bef7b","added_by":"auto","created_at":"2025-05-16 13:19:42","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":263999,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003emiRNA-mediated regulation of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e gene expression. An \u003c/strong\u003eOverlapanalysis of miRNAs targeting \u003cem\u003ePDK3\u003c/em\u003e was conducted via three different databases, andthe results were visualized via the InteractiVenn tool. \u003cstrong\u003eB \u003c/strong\u003e11 miRNAs regulating \u003cem\u003ePDK3\u003c/em\u003e expression with their miRDB target score, generated via SRplot. \u003cstrong\u003eC \u003c/strong\u003eExpression levels of all 11 miRNAs regulating \u003cem\u003ePDK3\u003c/em\u003e in different cancers.\u003c/p\u003e","description":"","filename":"Fig.3.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/4248c995c3ce5a9c568a582d.jpg"},{"id":82897314,"identity":"6f152f21-496d-4980-9a2c-52738cb275eb","added_by":"auto","created_at":"2025-05-16 13:03:42","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":561792,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAnalysis of genes coexpressed with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A \u003c/strong\u003eCoexpression analysis of \u003cem\u003ePDK3\u003c/em\u003e via STRING. \u003cstrong\u003eB\u003c/strong\u003eCoexpression analysis of \u003cem\u003ePDK3\u003c/em\u003e via GeneMANIA. \u003cstrong\u003eC \u003c/strong\u003eCoexpression analysis of \u003cem\u003ePDK3\u003c/em\u003e viaBioGRID. \u003cstrong\u003eD \u003c/strong\u003eOverlapanalysis of coexpressedgenes of \u003cem\u003ePDK3\u003c/em\u003e visualized through the InteractiVenn tool. \u003cstrong\u003eE \u003c/strong\u003eLollipop plot of \u003cem\u003ePDK3\u003c/em\u003e showing the correlation of \u003cem\u003ePDK3\u003c/em\u003e with different cancer types.\u003cstrong\u003e F \u003c/strong\u003eHallmark enrichment plot of \u003cem\u003ePDK3\u003c/em\u003e coexpressedgenes.\u003c/p\u003e","description":"","filename":"Fig.4.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/495192c89dda541a5fe174de.jpg"},{"id":82899508,"identity":"7037c2e4-3045-49f1-b932-2ed7a044a072","added_by":"auto","created_at":"2025-05-16 13:19:42","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":200608,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eImmune interactions of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A \u003c/strong\u003e\u003cem\u003ePDK3\u003c/em\u003e expression in different immune subtypes visualized in the TISIDB database. \u003cstrong\u003eB \u003c/strong\u003eHeatmap showing the correlation between \u003cem\u003ePDK3\u003c/em\u003e expression and immune cells in different cancer types retrieved from the TIMER 2.0 database. \u003cstrong\u003eC\u0026amp;D \u003c/strong\u003eBubble plot showing the correlation between \u003cem\u003ePDK3\u003c/em\u003e expression and anticancer drug sensitivity from the GSCA database.\u003c/p\u003e","description":"","filename":"Fig.5.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/5201fe52a26bdcf6c4dc965a.jpg"},{"id":82898462,"identity":"bbadbca8-ba47-45fe-a22d-3acf5a92ba39","added_by":"auto","created_at":"2025-05-16 13:11:42","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":127774,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eshRNA-mediated \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e knockdown in SiHa cells. \u003c/strong\u003eqRT‒PCR of colonies from transfected cells \u003cem\u003ein which PDK3 was knocked down\u003c/em\u003e. \u003cstrong\u003eB\u003c/strong\u003e Western blot analysis of colonies from transfected cells with \u003cem\u003ePDK3\u003c/em\u003e knockdown. \u003cstrong\u003eC\u003c/strong\u003eQuantitative analysis of the western blot data shown in the panel. \u003cstrong\u003eD \u003c/strong\u003eThe graph shows the rate of proliferation of scrambled and \u003cem\u003ePDK3\u003c/em\u003e k/d SiHa cells. \u003cstrong\u003eE\u003c/strong\u003eRepresentative images of the colony formation assay.\u003cstrong\u003e F \u003c/strong\u003eThe bar graph shows the results of the quantitative analysis of the colony formation assay presentedin the panel. The graph represents the mean and SD * p ≤ 0.05; **p ≤ 0.01 indicatesstatistical significance.\u003c/p\u003e","description":"","filename":"Fig.6.tif.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/eee65657304df70f7a95e6b2.jpg"},{"id":82901762,"identity":"84279ba9-cc0d-45de-a9e2-45e74fe90c0a","added_by":"auto","created_at":"2025-05-16 13:35:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3350190,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/85019d48-2742-4505-bedc-6aeaedf35b9b.pdf"},{"id":82899511,"identity":"161b4744-f21f-4aee-8da8-e28cd90e4794","added_by":"auto","created_at":"2025-05-16 13:19:43","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":479218,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary Fig. 1 \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e promoter DNA methylation and correlation between \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e expression and methylation. A \u003c/strong\u003eDifferential methylation of the \u003cem\u003ePDK3\u003c/em\u003epromoter visualized using UALCAN\u003cstrong\u003e B \u003c/strong\u003eHeatmap of \u003cem\u003ePDK3\u003c/em\u003emethylation at different CpG sites in all 3 cancers analysed using TCGA datasets from the UCSC Xena browser and plotted using an SR plot. \u003cstrong\u003eC \u003c/strong\u003eDifferential methylation levels of \u003cem\u003ePDK3\u003c/em\u003e observed in different cancers on considering CpGs within 1 kb upstream of the \u003cem\u003ePDK3\u003c/em\u003epromoter. \u003cstrong\u003eD \u003c/strong\u003eSpearman's correlation between \u003cem\u003ePDK3\u003c/em\u003e expression and promoter methylation was analysed using TCGA datasets from the UCSC Xena browser and plotted using an SR plot.\u003c/p\u003e","description":"","filename":"SuplementaryFig.1.tif","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/ce30e439dfed84da4d7a7503.tif"},{"id":82897324,"identity":"723fa5d4-59fb-4936-a523-f07649735ed4","added_by":"auto","created_at":"2025-05-16 13:03:42","extension":"tif","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":1633806,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary Fig. 2 Functional enrichment analysis of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eupstream transcriptional regulators.\u003c/strong\u003e \u003cstrong\u003eA \u003c/strong\u003eKEGG pathway analysis. \u003cstrong\u003eB \u003c/strong\u003eBiological processes. \u003cstrong\u003eC \u003c/strong\u003eMolecular processes. \u003cstrong\u003eD \u003c/strong\u003eCellular components.\u003c/p\u003e","description":"","filename":"SuplementaryFig.2.tif","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/cccca88cf373b751a2cde608.tif"},{"id":82901225,"identity":"22b904cd-c086-4e25-831c-9d3d7aad7791","added_by":"auto","created_at":"2025-05-16 13:27:42","extension":"tif","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":466394,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary Fig. 3 Overall survival and stagewise expression of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePDK3\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. An \u003c/strong\u003eOverall survival analysis of \u003cem\u003ePDK3\u003c/em\u003ebased on gene expression in different cancers plotted using KM plotter. \u003cstrong\u003eB\u003c/strong\u003e \u003cem\u003ePDK3\u003c/em\u003e gene expression in different clinical stages of cancer visualized using GEPIA database.\u003c/p\u003e","description":"","filename":"SuplementaryFig.3.tif","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/e56e594ed5d0f5f30371081d.tif"},{"id":82898458,"identity":"bd751b24-ac29-45bc-9c73-b0a0b3e3b465","added_by":"auto","created_at":"2025-05-16 13:11:42","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":34474,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary Table 1 \u003c/strong\u003eTranscription factors that interactwith \u003cem\u003ePDK3\u003c/em\u003e were obtained from the TFlink database.\u003c/p\u003e","description":"","filename":"SuplementaryTable.1.docx","url":"https://assets-eu.researchsquare.com/files/rs-6429535/v1/6e399db570ed37ff150e58f8.docx"}],"financialInterests":"","formattedTitle":"Understanding the Role of Pyruvate Dehydrogenase Kinase 3 in Gynecological Cancers","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe incidence of gynecological cancer (GC), a cancer affecting multiple organs of the female reproductive system, has substantially increased, contributing to 15.25% of all cancer cases reported and 15.77% of cancer-related deaths among women worldwide [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The global burden of gynecological cancer is predicted to increase by 49.61%, and the associated mortality is proposed to increase to 77.23% by the year 2050 [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Among all gynecological cancers, breast cancer continues to be the primary cause of cancer-associated death, followed by cervical cancer, ovarian cancer and endometrial cancer [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Metabolic reprogramming is an important hallmark of cancer, supporting the energy demand of rapidly proliferating and growing cancer cells. One such metabolic rewiring is the sustenance of a higher rate of glycolysis despite having sufficient oxygen, termed the Warburg effect [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Cancer cells have been shown to display rewired metabolic pathways, perhaps shifting away from oxidative phosphorylation and toward glycolysis.\u003c/p\u003e \u003cp\u003eThe pyruvate dehydrogenase kinase (PDK) family of proteins regulates the pyruvate dehydrogenase complex (PDHC), a critical regulatory enzyme that links glycolysis with the citric acid cycle. By controlling this complex, PDK plays a pivotal role in shifting cellular respiration toward the Warburg effect [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The PDK family consists of four isoforms, namely, PDK1, PDK2, PDK3, and PDK4, which share 70% homology and exhibit differential tissue expression, all of which are present in the mitochondrial matrix [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. PDK inactivates PDHC by phosphorylating serine residues at three different sites (Ser293, Ser300, and Ser232) in its E1α subunit [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. The inactivation of PDHC results in altered metabolic pathways, which may shift away from oxidative phosphorylation and toward glycolysis, supporting the energy demand of rapidly proliferating and growing cancer cells [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe activity of PDK is proportional to its binding efficiency to the lipoyl domain of the E2 subunit in PDH. Among all the isoforms, pyruvate dehydrogenase kinase 3 (PDK3) has the highest affinity and is not affected by the presence of the substrate pyruvate [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Recent findings indicate a multifaceted role of \u003cem\u003ePDK3\u003c/em\u003e in cancer progression [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. \u003cem\u003ePDK3\u003c/em\u003e is upregulated in cancers such as colon [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], acute myeloid leukemia [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], gastric [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] and urothelial carcinomas [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003ePDK3\u003c/em\u003e promotes tumorigenesis by stimulating cell proliferation, angiogenesis, invasion, and glycolysis in multiple cancer types [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Immunohistochemical staining has shown a direct correlation between \u003cem\u003ePDK3\u003c/em\u003e expression and disease severity in colon cancer cells [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. \u003cem\u003ePDK3\u003c/em\u003e overexpression reportedly contributes to chemoresistance and cancer recurrence through the combined action of Hypoxia-Inducing Factor α (\u003cem\u003eHIF-α\u003c/em\u003e) and Heat Shock Factor 1(\u003cem\u003eHSF1\u003c/em\u003e) [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. \u003cem\u003ePDK3\u003c/em\u003e also regulates the PI3K-Akt pathway to promote tumorigenic characteristics [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. \u003cem\u003ePDK3\u003c/em\u003e inhibits the T-cell-mediated immune response and suppresses the expression of Programmed Death Ligand-2 (PDL-2) by inhibiting the Jak1/STAT 1 pathway, thereby promoting cancer growth [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. \u003cem\u003ePDK3\u003c/em\u003e could be used as a potential target in cancer because of its multifaceted tumor-promoting functions.\u003c/p\u003e \u003cp\u003eNevertheless, the significance of \u003cem\u003ePDK3\u003c/em\u003e in gynecological cancers remains poorly understood. To address this gap, we carried out a comprehensive analysis to understand the role of \u003cem\u003ePDK3\u003c/em\u003e in gynecological cancers. We systematically studied the expression, regulation, clinical utility, interaction networks, biological functions, immune infiltration, and chemical activity of PDK3 to target \u003cem\u003ePDK3\u003c/em\u003e in gynecological cancers. Finally, we performed shRNA-mediated \u003cem\u003ePDK3\u003c/em\u003e knockdown studies to assess the role of \u003cem\u003ePDK3\u003c/em\u003e in cervical cancer.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eData acquisition and processing\u003c/h2\u003e \u003cp\u003e \u003cem\u003ePDK3\u003c/em\u003e expression data were sourced from the Genotype-Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA) databases [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The GTEx database offers a detailed catalogue of gene expression levels among various tissue types, facilitating the comparison of \u003cem\u003ePDK3\u003c/em\u003e expression status in different tissue types. Box plots showing the differential expression of \u003cem\u003ePDK3\u003c/em\u003e between normal and tumor samples were generated via the TIMER2.0 database under the \u0026lsquo;Gene DE\u0026rsquo; module [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The genomic, transcriptomic, and clinical data from TCGA were retrieved via UCSC Xena and GEPIA [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. \u003cem\u003ePDK3\u003c/em\u003e expression between tumors and normal tissues was tested for significance via the Wilcoxon rank-sum test. Statistical significance was set at a p value of \u0026lt;\u0026thinsp;0.05. The SRPlot tool was used to represent the obtained result in box plot form [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Furthermore, \u003cem\u003ePDK3\u003c/em\u003e expression data were cross-validated with datasets from the Cancer Cell Line Encyclopedia (CCLE) [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eExploration of genetic and epigenetic alterations\u003c/h3\u003e\n\u003cp\u003eThe IntOGen [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] database was analysed to retrieve mutations connected with \u003cem\u003ePDK3\u003c/em\u003e in the pancancer samples. The TCGA database was used to visualize mutations in different cancer types. The TIMER 2.0 platform was used to visualize the frequency and types of mutations. Additionally, copy number variations (CNVs), amplifications, and deletions linked to \u003cem\u003ePDK3\u003c/em\u003e were examined via the GSCA database [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. We next analysed the association between DNA methylation and \u003cem\u003ePDK3\u003c/em\u003e expression. \u003cem\u003ePDK3\u003c/em\u003e promoter DNA methylation in normal and tumor samples was tested via the UALCAN database [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. We specifically selected CpG sites located within 1000 base pairs upstream of the \u003cem\u003ePDK3\u003c/em\u003e promoter. The correlation between methylation and expression was tested via Spearman correlation analysis [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo explore the post-transcriptional regulation of \u003cem\u003ePDK3\u003c/em\u003e, we performed an extensive search of miRNAs that target \u003cem\u003ePDK3\u003c/em\u003e via three miRNA target scanning tools, namely, miRNet [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], miRDB [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], and TargetScan [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. The common miRNAs found in these three databases were identified via the IteractiVenn tool [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eCoexpression and protein‒protein interaction network construction\u003c/h3\u003e\n\u003cp\u003eWe used the GeneMANIA [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] and BioGRID databases [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] to construct the \u003cem\u003ePDK3\u003c/em\u003e and its coexpressed gene network. The STRING web server was used to construct the protein‒protein interaction network among coexpressed genes [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Functional linkages and interactions involving \u003cem\u003ePDK3\u003c/em\u003e were interpreted by combining the interaction data with correlation coefficients from the TIMER2.0 database, which focused on gynecological cancer types. The Cancerhallmarks [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] tool was used to visualize the enriched cancer hallmarks related to \u003cem\u003ePDK3\u003c/em\u003e coexpressed genes. This method allows for a thorough examination of \u003cem\u003ePDK3\u003c/em\u003e's regulatory network and possible contribution to the development of cancer.\u003c/p\u003e\n\u003ch3\u003eTranscriptional regulator identification and functional enrichment analysis\u003c/h3\u003e\n\u003cp\u003eTFlink [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] was used to identify the transcription factors regulating \u003cem\u003ePDK3\u003c/em\u003e. By combining information from chromatin immunoprecipitation (ChIP) investigations with computational predictions, TFlink offers information on putative transcription factors that may control genes of interest. The biological pathways linked to these regulators and the enrichment of gene ontology (GO) terms were evaluated using ShinyGO [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eClinical implications and survival outcomes\u003c/h3\u003e\n\u003cp\u003eThe correlation between \u003cem\u003ePDK3\u003c/em\u003e expression and cancer progression stage was determined via the GDC-TCGA datasets accessible via GEPIA [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. The prognostic significance of \u003cem\u003ePDK3\u003c/em\u003e expression was evaluated via Kaplan‒Meier plotter for survival analysis [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eImmune infiltration and drug interaction analysis\u003c/h2\u003e \u003cp\u003eThe TISIDB tumor-immune system interaction database was used to analyse immune cell infiltration patterns and \u003cem\u003ePDK3\u003c/em\u003e expression [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. The relationships between \u003cem\u003ePDK3\u003c/em\u003e expression and 24 distinct tumor-infiltrating lymphocyte subsets were examined via the TIMER 2.0 program. SRplot was used to create heatmaps from the data. Finally, the GSCA database was used to examine the relationship between \u003cem\u003ePDK3\u003c/em\u003e expression and drug response [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Data on drug‒gene correlations from the Cancer Therapeutics Response Portal (CTRP) and Genomics of Drug Sensitivity in Cancer (GDSC) databases are integrated by the GSCA. SRplot was used to create a bubble plot that represented the data. The various databases and their links used in the present study are listed in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eCell culture and transfection\u003c/h3\u003e\n\u003cp\u003eSiHa cells were cultured and maintained according to ATCC guidelines. To generate the \u003cem\u003ePDK3\u003c/em\u003e knockdown cell line, SiHa cells (2\u0026times;10\u003csup\u003e6\u003c/sup\u003e) were seeded in a 6-well plate and incubated at 37\u0026deg;C in a 5% CO\u003csub\u003e2\u003c/sub\u003e environment. After 24 hours, the cells were transfected with a shRNA plasmid against \u003cem\u003ePDK3\u003c/em\u003e (sc-39029-SH; Santa Cruz). Control plasmid (sc-108060; Santa Cruz)-transfected cells served as controls. We used Lipofectamine 3000 (Thermo Fisher Scientific, USA) to carry out the transfections according to the manufacturer's protocol. \u003cem\u003ePDK3\u003c/em\u003e-knockdown clones were isolated by culturing the cells in 2.0 \u0026micro;g/ml puromycin for 14 days [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eConfirmation of\u003c/b\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eknockdown\u003c/b\u003e\u003c/p\u003e \u003cp\u003eTotal RNA was isolated using TRIzol reagent (Thermo Fisher Scientific, USA). \u003cem\u003ePDK3\u003c/em\u003e Hs00178440_m1; Invitrogen) knockdown was confirmed by TaqMan-based qRT-PCR using 25ng of RNA with β-actin as an endogenous control. Gene expression was calculated using the ΔΔCt method, where ΔCt\u0026thinsp;=\u0026thinsp;Ct (target gene) - Ct (reference gene), ΔΔCt\u0026thinsp;=\u0026thinsp;ΔCt test - ΔCt control. The fold change (FC) was calculated as FC\u0026thinsp;=\u0026thinsp;2^(-ΔΔCt) [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAfter being washed in ice-cold PBS, \u003cem\u003ePDK3\u003c/em\u003e knockdown and vector-transfected cells were lysed in RIPA buffer containing a protease inhibitor. Using SDS‒PAGE, approximately 40 \u0026micro;g of protein was separated and then blotted onto a nitrocellulose membrane. The membranes were incubated overnight at 4\u0026deg;C in the presence of primary antibodies against PDK3 (1:2000 dilution, ABclonal, USA) and anti-ACTB (1:5000 dilution, ABclonal, USA). The membranes were then incubated for two hours at room temperature with an HRP-conjugated secondary antibody after being washed with TBST. To visualize the PDK3 protein, Clarity Western ECL Substrate (Bio-Rad, USA) was utilized. An iBright imaging system (Invitrogen, USA) was used to capture images [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eProliferation assay\u003c/h3\u003e\n\u003cp\u003eTrypsinization and trypan blue staining were performed on the cells every 24 hours, and a Neubauer chamber was used to count living cells under a microscope. Omni calculator was used to determine the doubling times of the control and knockdown cells [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eColony formation assay\u003c/h2\u003e \u003cp\u003eApproximately 500 cells were cultivated in a 6 cm cell culture plate for two weeks in full media. Following the disposal of the culture media and washing with PBS, the colonies were stained for ten to fifteen minutes with 0.5% crystal violet in methanol staining solution. After extra stain was removed and the samples were washed with PBS, the number of stained colonies was calculated [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eis overexpressed in gynecological cancers\u003c/b\u003e\u003c/p\u003e \u003cp\u003eUsing the GTEX database, we first examined the expression of \u003cem\u003ePDK3\u003c/em\u003e in several tissue types. \u003cem\u003ePDK3\u003c/em\u003e was expressed at varying levels in various human tissues (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). Pancancer analysis of \u003cem\u003ePDK3\u003c/em\u003e expression via the TIMER 2.0 tool revealed that \u003cem\u003ePDK3\u003c/em\u003e was upregulated in tumor samples compared with normal samples in the TCGA cohort (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). We found that \u003cem\u003ePDK3\u003c/em\u003e mRNA levels were significantly elevated in uterine corpus endometrial carcinoma (TCGA-UCEC), cervical squamous cell carcinoma (TCGA-CESC), and invasive breast carcinoma (TCGA-BRCA) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). Interestingly, \u003cem\u003ePDK3\u003c/em\u003e expression did not significantly change in ovarian cancer (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD). High expression of \u003cem\u003ePDK3\u003c/em\u003e was observed in different gynecological cell lines (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eE).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eRegulation of\u003c/b\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eacross gynecological cancers\u003c/b\u003e\u003c/p\u003e \u003cp\u003eMutation analysis via IntOGen identified 129 mutations in \u003cem\u003ePDK3\u003c/em\u003e, with missense mutations being the most frequent (60%), followed by synonymous (29%), truncating (8%), and splice-site (3%) mutations. Hotspot mutations were detected, especially in the functional domains of \u003cem\u003ePDK3\u003c/em\u003e, indicating that these mutations might affect the function of \u003cem\u003ePDK3\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). According to TCGA dataset analysis, mutations in \u003cem\u003ePDK3\u003c/em\u003e were absent in breast cancer patients, 0.68% in cervical cancer patients, and slightly more frequently (6.4%) in endometrial cancer patients (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). The low mutation prevalence indicates that although genetic modifications do occur, the total mutation rate across tumors is still low, suggesting that \u003cem\u003ePDK3\u003c/em\u003e dysregulation in the TCGA-CESC (0.68%) and TCGA-BRCA (0%) cohorts might originate from other mechanisms, such as DNA methylation or miRNA (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo investigate factors other than mutations that potentially regulate the expression of \u003cem\u003ePDK3\u003c/em\u003e, we examined copy number variations (CNVs) in endometrial, cervical, and breast malignancies. Remarkably, a sizable portion of patients presented gains at the \u003cem\u003ePDK3\u003c/em\u003e locus; amplifications were detected in almost 17% of patients with breast cancer, 16% of patients with cervical cancer, and approximately 14% of patients with endometrial cancer (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Linking these CNVs with \u003cem\u003ePDK3\u003c/em\u003e expression revealed that there was a positive correlation between these CNVs and \u003cem\u003ePDK3\u003c/em\u003e expression, especially in endometrial cancer (Spearman R coefficient 0.19, p value\u0026thinsp;\u0026lt;\u0026thinsp;0.05) and invasive breast carcinoma (Spearman R coefficient 0.09, p value\u0026thinsp;\u0026lt;\u0026thinsp;1001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE). Cervical cancer had a Spearman R coefficient of 0.02, suggesting that the upregulation of \u003cem\u003ePDK3\u003c/em\u003e in cervical cancer may be independent of CNV. These data indicate that CNVs may significantly contribute to the overexpression of \u003cem\u003ePDK3\u003c/em\u003e in breast cancer.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe GSCA database showing various percentages and types of SNVs and CNVs at the PDK3 locus in different gynaecological cancers.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e \u003cp\u003ePercentages and types of SNVs and CNVs at the PDK3 locus in different gynecological cancers\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e \u003cp\u003eSNVs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCancer type\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eEffective mutations\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNon effective mutations\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eSample size\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePercentage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBRCA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e1026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCESC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e291\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.687285223\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eUCEC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e531\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.779661017\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"7\" nameend=\"c7\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCNVs\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCancer type\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eTotal amplification\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003eDeletion\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eHeterozygous amplification\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eHeterozygous deletion\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eHomozygous amplification (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003eHomozygous deletion\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBRCA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.2222222\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.6481481\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.1296296\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.5740741\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37037\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCESC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e16.6101695\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21.6949153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.2372881\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.6949153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.3728814\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eUCEC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.1001855\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.7699443\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.1001855\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15.3988868\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.371058\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAdditionally, we examined the effects of epigenetic changes by evaluating DNA methylation patterns at the \u003cem\u003ePDK3\u003c/em\u003e promoter in several gynecological malignancies (Supplementary Fig.\u0026nbsp;1A). Our analysis revealed significant methylation at most of the CpG sites (p value\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in breast cancer and uterine cancer (Supplementary Fig.\u0026nbsp;1B) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Promoter methylation analysis revealed that the promoter was not hypermethylated despite a significant increase in methylation (a beta value cut-off of 0.7\u0026ndash;0.5 indicated hypermethylation, and 0.3\u0026ndash;0.25 indicated hypomethylation), suggesting that additional epigenetic elements, such as miRNAs, are involved in regulating gene expression (Supplementary Fig.\u0026nbsp;1C). Correlation analysis revealed that \u003cem\u003ePDK3\u003c/em\u003e expression and promoter methylation were strongly negatively correlated in the TCGA-BRCA cohort and TCGA-CESC cohort (Supplementary Fig.\u0026nbsp;1D). These findings suggest that different epigenetic factors play a role in controlling the expression of \u003cem\u003ePDK3\u003c/em\u003e in individual cancer types.\u003c/p\u003e \n\u003cp\u003e\u003cstrong\u003eTable 2\u0026nbsp;\u003c/strong\u003eThe OncoDB database showing methylation levels of PDK3 observed on CpGs in different cancers. (Probes in red colour shows the significant differential methylation).\u003c/p\u003e\n\u003ctable style=\"width:101.08%;background:white;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:21.78%;border:inset #333333 1.0pt;background:#D1D1FB;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003eProbe\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:12.26%;border:inset #333333 1.0pt;border-left:none;background:#D1D1FB;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ePosition\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:23.48%;border:inset #333333 1.0pt;border-left:none;background:#D1D1FB;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003eAverage of Cancer Sample\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border:inset #333333 1.0pt;border-left: none;background:#D1D1FB;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003eAverage of Normal Sample\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border:inset #333333 1.0pt;border-left: none;background:#D1D1FB;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ep-value\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\" style=\"width:100.0%;border:inset #333333 1.0pt;border-top:none;background:#D9D9D9;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003eBRCA\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg20258741\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24464729\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.41\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.44\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e4.7e-06\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg23143524\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464960\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.55\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.54\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e2.6e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg02539591\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464975\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.41\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.42\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e2.0e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg02623256\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24465114\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.39\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.39\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e8.3e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg06721573\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465117\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.40\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.44\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e6.2e-04\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg01212677\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465173\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.46\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.50\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e2.1e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg15308487\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465175\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.31\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.33\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.8e-02\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg15345580\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465177\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.23\u003c/span\u003e\u003c/p\u003e\n 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\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg22969661\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24465359\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.51\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.51\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e9.8e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg00114625\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465361\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.55\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.56\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e8.0e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg18414950\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465779\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.48\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.58\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e2.0e-12\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg25863147\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24466244\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.47\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.57\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.6e-11\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg13777527\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24539503\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.87\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.94\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.6e-12\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\" style=\"width:100.0%;border:inset #333333 1.0pt;border-top:none;background:#D9D9D9;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003eCESC\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg20258741\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464729\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.45\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.38\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e2.2e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg23143524\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464960\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.58\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.47\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e1.3e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg02539591\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464975\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.44\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.37\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e3.4e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg02623256\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24465114\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.43\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.38\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e3.5e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg06721573\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465117\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.41\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.47\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e5.0e-07\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg01212677\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24465173\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.48\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.44\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e3.3e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n 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style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.30\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e4.2e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg15345580\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465177\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.28\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.14\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.0e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg22969661\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465359\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.53\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.43\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e9.1e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg00114625\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465361\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.55\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.51\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e2.1e-09\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg18414950\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24465779\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.53\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.51\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e8.4e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg25863147\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24466244\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.61\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.67\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n 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style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.89\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.94\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e3.6e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\" style=\"width:100.0%;border:inset #333333 1.0pt;border-top:none;background:#D9D9D9;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003eUCEC\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg20258741\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464729\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.43\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.42\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e2.5e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg23143524\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24464960\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.57\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.53\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e8.6e-04\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg02539591\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24464975\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.41\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.41\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n 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style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465117\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.39\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.42\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e3.9e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg01212677\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465173\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.48\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.51\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e3.9e-02\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg15308487\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465175\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.33\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.36\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.7e-02\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg15345580\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465177\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.25\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.21\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e6.5e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg22969661\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465359\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.53\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.50\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e2.2e-03\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg00114625\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465361\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.56\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.54\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e2.1e-02\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg18414950\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24465779\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.51\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.55\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.7e-05\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003ecg25863147\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e24466244\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.69\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e0.68\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:#383838;'\u003e6.3e-01\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width:22.32%;border:inset #333333 1.0pt;border-top:none;background:transparent;padding:.75pt .75pt .75pt .75pt;height:1.9pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003ecg13777527\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:11.84%;border-top:none;border-left: none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;height:1.9pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e24539503\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.36%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;height:1.9pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.87\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:23.34%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;height:1.9pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e0.93\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:19.14%;border-top:none;border-left:none;border-bottom:inset #333333 1.0pt;border-right:inset #333333 1.0pt;background:transparent;padding:.75pt .75pt .75pt .75pt;height:1.9pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Aptos\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-family:\"Calibri\",sans-serif;color:red;'\u003e1.6e-12\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTable showing all the databases and tools used in the current study.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDatabases/Sites used in the study\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLinks\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGenotype-Tissue Expression (GTEx)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://gtexportal.org/home/\u003c/span\u003e\u003cspan address=\"https://gtexportal.org/home/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTIMER2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://timer.cistrome.org/\u003c/span\u003e\u003cspan address=\"http://timer.cistrome.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUCSC Xena\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://xena.ucsc.edu/\u003c/span\u003e\u003cspan address=\"https://xena.ucsc.edu/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUALCAN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://ualcan.path.uab.edu/\u003c/span\u003e\u003cspan address=\"https://ualcan.path.uab.edu/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCancer Cell Line Encyclopedia (CCLE)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://sites.broadinstitute.org/ccle/\u003c/span\u003e\u003cspan address=\"https://sites.broadinstitute.org/ccle/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOncoDB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://oncodb.org/\u003c/span\u003e\u003cspan address=\"https://oncodb.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGene Set Cancer Analysis (GSCA)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://guolab.wchscu.cn/GSCA/#/\u003c/span\u003e\u003cspan address=\"https://guolab.wchscu.cn/GSCA/#/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGeneMANIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://genemania.org/\u003c/span\u003e\u003cspan address=\"https://genemania.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBioGRID\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://thebiogrid.org/\u003c/span\u003e\u003cspan address=\"https://thebiogrid.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSTRING\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://string-db.org/\u003c/span\u003e\u003cspan address=\"https://string-db.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCancer hallmarks\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://cancerhallmarks.com/\u003c/span\u003e\u003cspan address=\"https://cancerhallmarks.com/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTFlink\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://tflink.net/\u003c/span\u003e\u003cspan address=\"https://tflink.net/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGEPIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://gepia.cancer-pku.cn/\u003c/span\u003e\u003cspan address=\"http://gepia.cancer-pku.cn/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKaplan-Meier plotter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://kmplot.com/analysis/\u003c/span\u003e\u003cspan address=\"https://kmplot.com/analysis/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTISIDB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://cis.hku.hk/TISIDB/\u003c/span\u003e\u003cspan address=\"http://cis.hku.hk/TISIDB/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntOGen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.intogen.org/search\u003c/span\u003e\u003cspan address=\"https://www.intogen.org/search\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSRplot\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.bioinformatics.com.cn/srplot\u003c/span\u003e\u003cspan address=\"https://www.bioinformatics.com.cn/srplot\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVenny 2.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://bioinfogp.cnb.csic.es/tools/venny/\u003c/span\u003e\u003cspan address=\"https://bioinfogp.cnb.csic.es/tools/venny/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShinyGO 0.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://bioinformatics.sdstate.edu/go/\u003c/span\u003e\u003cspan address=\"https://bioinformatics.sdstate.edu/go/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emiRNET\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.mirnet.ca/upload/MirUploadView.xhtml\u003c/span\u003e\u003cspan address=\"https://www.mirnet.ca/upload/MirUploadView.xhtml\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emiRDB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://mirdb.org/\u003c/span\u003e\u003cspan address=\"https://mirdb.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTargetScan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.targetscan.org/vert_80/\u003c/span\u003e\u003cspan address=\"https://www.targetscan.org/vert_80/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCell doubling time calculator\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.omnicalculator.com/\u003c/span\u003e\u003cspan address=\"https://www.omnicalculator.com/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eWe investigated the role of microRNAs (miRNAs) as potential regulators of \u003cem\u003ePDK3\u003c/em\u003e expression. Using data from the miRDB, miRNET and TargetScan databases, we identified 11 miRNAs that could target \u003cem\u003ePDK3\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA) with different target scores (a score of 70\u0026ndash;100 indicating high confidence) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Among the 11 miRNAs, hsa-miR-525 and hsa-miR-369 were downregulated in all three cancers, and hsa-miR-186 and hsa-miR-494 were downregulated only in TCGA-BRCA and TCGA-CESC, respectively. hsa-miR-205 and hsa-miR-194 were downregulated only in the TCGA-BRCA cohort and TCGA-CESC cohort, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). Hence, downregulation of these miRNAs might reduce their suppressive effect on the \u003cem\u003ePDK3\u003c/em\u003e gene.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eCo-expression and interaction analysis of\u003c/b\u003e \u003cb\u003ePDK3\u003c/b\u003e\u003c/p\u003e \u003cp\u003eUsing GeneMANIA (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA) and BioGRID (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB), we constructed a coexpression network of \u003cem\u003ePDK3\u003c/em\u003e. The STRING tool was used to generate the PPIN (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC). Protein‒protein interaction (PPI) network analysis revealed a complex network consisting of 14 nodes and 34 edges. The network demonstrated significant PPI enrichment, indicating nonrandom interactions among the identified proteins. Furthermore, the protein‒protein interaction network (PPIN) constructed from the overlapping genes revealed a direct interaction between \u003cem\u003ePDK3\u003c/em\u003e and 8 members (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD). Thus, our \u003cem\u003ein silico\u003c/em\u003e analysis revealed strong interactions of PDK3 with DLAT, DLD, PDHA1, PDHB, PDHX, PDK1, PDK2, and PDK4. This finding was supported by correlation analysis, which revealed positive correlations between \u003cem\u003ePDK3\u003c/em\u003e and DLAT, DLD, PDHA1, PDHB, PDHX, and PDK1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE). Furthermore, a cancer hallmark enrichment plot of \u003cem\u003ePDK3\u003c/em\u003e coexpressed genes revealed significant involvement in reprogramming and energy metabolism (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eF).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003emodulates critical signalling pathways in cancer progression\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eIn silico\u003c/em\u003e analysis revealed \u003cem\u003ePDK3\u003c/em\u003e as part of many key oncogenic signalling pathways. We used ChIP-seq data available at TFlink to identify key transcription factors (TFs) regulating \u003cem\u003ePDK3\u003c/em\u003e expression. We determined that \u003cem\u003ePDK3\u003c/em\u003e possesses binding sites close to 414 transcription factors (Supplementary Table\u0026nbsp;1). The upstream regulators of \u003cem\u003ePDK3\u003c/em\u003e are predominantly connected with pathways in cancer, the cell cycle, Wnt signalling, miRNAs in cancer, breast cancer, and HPV infection, as shown by KEGG pathway enrichment analysis (Supplementary Fig.\u0026nbsp;2A). Gene Ontology (GO) term analysis further suggested that enrichment of RNA biosynthesis, differential transcription, and macromolecule biosynthesis were the top hit BPs (Supplementary Fig.\u0026nbsp;2B). Significant enrichment in transcription coregulator activity and different transcription factor binding activities was observed for \u003cem\u003ePDK3\u003c/em\u003e targets at the molecular function (MF) level. The majority of these functions were linked to cellular components that regulate DNA epigenetically (Supplementary Fig.\u0026nbsp;2C \u0026amp; 2D).\u003c/p\u003e \u003cp\u003e \u003cb\u003eClinical significance of\u003c/b\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eacross gynecological cancers\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe clinical significance of aberrant \u003cem\u003ePDK3\u003c/em\u003e expression in gynecological cancers was evaluated via the GEPIA tool. Correlation analysis of \u003cem\u003ePDK3\u003c/em\u003e expression with overall survival revealed significant prognostic implications in breast and cervical cancer patients. Notably, increased \u003cem\u003ePDK3\u003c/em\u003e expression in the TCGA-CESC cohort and decreased PDK3 expression in the TCGA-BRCA and TCGA-UCEC cohorts correlated with improved survival outcomes (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) (Supplementary Fig.\u0026nbsp;3A). We next tested the association of \u003cem\u003ePDK3\u003c/em\u003e expression with cancer stage in breast, cervical, and endometrial cancer. There were no significant differences between \u003cem\u003ePDK3\u003c/em\u003e expression and cancer stage (F value\u0026thinsp;\u0026lt;\u0026thinsp;2.5) (Supplementary Fig.\u0026nbsp;3B).\u003c/p\u003e \u003cp\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eexpression can affect the abundance of immune cells\u003c/b\u003e\u003c/p\u003e \u003cp\u003eWe generated a violin plot via TISIDB to evaluate \u003cem\u003ePDK3\u003c/em\u003e expression in different immune subtypes, classifying immune subtypes into six classes: wound healing (C1), IFN-gamma dominant (C2), inflammatory (C3), lymphocyte depleted (C4), immunologically quiet (C5), and TGF-β dominant (C6). \u003cem\u003ePDK3\u003c/em\u003e expression correlated with five immunological subtypes (C1, C2, C3, C4 and C6) in the TCGA-BRCA cohort and three immunological subtypes (C1, C2, and C4) in the TCGA-CESC cohort, indicating that \u003cem\u003ePDK3\u003c/em\u003e expression differs according to the subtype of the immune microenvironment (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA). TIMER 2.0 database analysis revealed an inverse relationship between \u003cem\u003ePDK3\u003c/em\u003e expression and the numbers of NK cells and CD8\u0026thinsp;+\u0026thinsp;T cells in the TCGA-BRCA cohort. In the TCGA-CESC cohort, unpolarized memory B cells were positively correlated with \u003cem\u003ePDK3\u003c/em\u003e expression. Additionally, \u003cem\u003ePDK3\u003c/em\u003e expression is positively correlated with antitumour M1 macrophages and negatively correlated with protomeric M2 macrophages. In addition, \u003cem\u003ePDK3\u003c/em\u003e overexpression was significantly negatively correlated with the expression of memory B cells and NK cells. In the TCGA-UCEC cohort, a significant negative correlation between NK cells and \u003cem\u003ePDK3\u003c/em\u003e was observed (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003e-drug interactions\u003c/b\u003e \u003c/p\u003e \u003cp\u003eHigher \u003cem\u003ePDK3\u003c/em\u003e expression was positively correlated with the expression of most GDSC drugs and one of the CTRP drugs, dasatinib, indicating that increased expression of this gene may lead to resistance to these drugs. A negative correlation of \u003cem\u003ePDK3\u003c/em\u003e was observed with most of the CTRP drugs and two of the GDSC drugs, GSK690693 and PLX470, suggesting that increased expression of this gene might lead to drug sensitivity (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eC\u0026amp;D).\u003c/p\u003e \u003cp\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eknockdown in SiHa cells\u003c/b\u003e\u003c/p\u003e \u003cp\u003eWe previously reported that \u003cem\u003ePDK3\u003c/em\u003e is significantly overexpressed in cervical cancer patient samples (Srinath et al., 2024). This finding was further confirmed by \u003cem\u003ein silico\u003c/em\u003e differential expression analysis of the TCGA-CESC datasets. qRT‒PCR and western blot analysis revealed significant expression of \u003cem\u003ePDK3\u003c/em\u003e in SiHa cells. Hence, we knocked down \u003cem\u003ePDK3\u003c/em\u003e in SiHa cells to investigate its biological properties. \u003cem\u003ePDK3\u003c/em\u003e knockdown in SiHa cells was confirmed by qRT‒PCR, which revealed an 85% decrease in the expression of \u003cem\u003ePDK3\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA). The knockdown effect was then confirmed by western blot analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB \u0026amp; C).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ePDK3\u003c/b\u003e \u003cb\u003eknockdown inhibits the growth and proliferation of SiHa cells\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003ePDK3\u003c/em\u003e knockdown in SiHa cells was confirmed by qRT‒PCR and western blotting (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA‒6C). The impact of \u003cem\u003ePDK3\u003c/em\u003e on proliferation and growth was quantified via growth curve analysis and colony formation assays. The growth curve analysis revealed a significant reduction in cell number upon \u003cem\u003ePDK3\u003c/em\u003e knockdown in SiHa cells. In contrast, the scrambled control-treated cells exhibited significant cell proliferation (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eD). The cell doubling time in scrambled and \u003cem\u003ePDK3\u003c/em\u003e-knockdown cells was 39.27 hrs. and 173.45 hrs., respectively. We next investigated the impact of \u003cem\u003ePDK3\u003c/em\u003e knockdown on the growth of SiHa cells via a colony formation assay. We found that, compared with scrambled treatment, PDK3 knockdown significantly reduced the number of colonies (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eE and F).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe involvement of \u003cem\u003ePDK3\u003c/em\u003e in cancer progression has gained significant attention because of its critical role in regulating cellular metabolism, especially in inhibiting PDHC. In the present study, we investigated the regulation, functional implications, and clinical significance of aberrant \u003cem\u003ePDK3\u003c/em\u003e expression in gynecological cancers through \u003cem\u003ein silico\u003c/em\u003e analysis and \u003cem\u003ein vitro\u003c/em\u003e experiments. According to earlier research, \u003cem\u003ePDK3\u003c/em\u003e is overexpressed in the stomach [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e], colon [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], lung [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e], cervix [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e], breast [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], and urinary bladder [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] and may contribute to metabolic reprogramming and tumor progression [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Despite its aberrant expression in multiple cancers [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], its role in gynecological cancers has not been well investigated. Our \u003cem\u003ein silico\u003c/em\u003e analysis of the TCGA-BRCA, TCGA-UCEC, and TCGA-CESC datasets revealed notable overexpression of \u003cem\u003ePDK3\u003c/em\u003e in cancer samples compared with normal samples. The Cell Line Encyclopedia (CCLE) also confirmed relatively high \u003cem\u003ePDK3\u003c/em\u003e expression in gynecological cancer cell lines, further emphasizing the oncogenic potential of \u003cem\u003ePDK3\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eGenetic and epigenetic analyses of \u003cem\u003ePDK3\u003c/em\u003e revealed that somatic \u003cem\u003ePDK3\u003c/em\u003e mutations were relatively uncommon in all of the cancer samples examined. Considerable CNV gains were observed in the TCGA-CESC, TCGA-UCEC, and TCGA-BRCA samples. Spearman correlation analysis revealed a positive correlation of \u003cem\u003ePDK3\u003c/em\u003e expression only in the TCGA-UCEC cohort and TCGA-BRCA cohort. Hence, CNV gain may have contributed to \u003cem\u003ePDK3\u003c/em\u003e overexpression in the TCGA-UCEC and TCGA-BRCA cohorts but not in the TCGA-CESC cohort. This finding provides the scope for further investigation of the cause of \u003cem\u003ePDK3\u003c/em\u003e overexpression in the TCGA-CESC cohort.\u003c/p\u003e \u003cp\u003eAberrant DNA methylation has been shown to influence gene expression, with hypermethylation contributing to gene silencing and hypomethylation contributing to gene upregulation [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Although our DNA methylation studies revealed significantly increased methylation in the normal tissues of all three cancers at specific CpG sites, the findings were inconsistent across all the CpG sites. The beta value cut-off indicated that neither significant hypermethylation nor hypomethylation was present in the tumor samples. However, according to the Spearman correlation analysis, the TCGA-BRCA and TCGA-CESC cohorts presented significant negative correlations with promoter methylation and \u003cem\u003ePDK3\u003c/em\u003e expression. We previously reported that \u003cem\u003ePDK3\u003c/em\u003e is an upregulated gene in cervical cancer [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. More detailed studies are needed to understand the relationship between promoter DNA methylation and \u003cem\u003ePDK3\u003c/em\u003e expression.\u003c/p\u003e \u003cp\u003eMicroRNA interaction and regulatory network analyses have identified multiple microRNAs that possibly target \u003cem\u003ePDK3\u003c/em\u003e expression in gynecological cancers. Importantly, hsa-miR-186-5p, hsa-miR-194-5p, hsa-miR-7-5p and hsa-miR-494-3p have been shown to target \u003cem\u003ePDK3\u003c/em\u003e. hsa-miR-186-5p downregulates \u003cem\u003ePDK3\u003c/em\u003e expression in glioma [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e], whereas hsa-7-5p has been studied for its role in suppressing \u003cem\u003ePDK3\u003c/em\u003e expression in TCGA-HNSC [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Our previous research revealed that hsa-miR-494-3p targets \u003cem\u003ePDK3\u003c/em\u003e in cervical cancer and that the binding of \u003cem\u003ePDK3\u003c/em\u003e by hsa-miR-494-3p results in the reversal of tumorigenic properties through metabolic reprogramming [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. This differential miRNA regulation suggests that the diverse miRNAs act to tightly regulate \u003cem\u003ePDK3\u003c/em\u003e expression among different cancer types to accommodate distinct metabolic adaptations.\u003c/p\u003e \u003cp\u003eCoexpression analysis of \u003cem\u003ePDK3\u003c/em\u003e highlighted the potential interaction of \u003cem\u003ePDK3\u003c/em\u003e with PDK1, PDK2, PDK4, PDHX, PDHB, PDHA1, DLAT, and DLD. The interaction of \u003cem\u003ePDK3\u003c/em\u003e with genes such as PDHX, PDHB, DLAT and DLD may be important for promoting metabolic reprogramming and requires detailed investigation. The hallmark enrichment analysis highlighted the role of PDK in metabolic reprogramming in cancer cells. Hence, more thorough mechanistic research is needed to understand how \u003cem\u003ePDK3\u003c/em\u003e and its coexpressed genes contribute to metabolic reprogramming and the development of cancer. Prognostic markers are clinical indicators used in oncology to determine a patient's risk of disease recurrence following initial therapy. Previous studies have shown that \u003cem\u003ePDK3\u003c/em\u003e influences patient survival in urothelial carcinoma [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], colon cancer [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], and cholangiocarcinoma [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Thus, we explored the prognostic significance of \u003cem\u003ePDK3\u003c/em\u003e in breast, cervical, and endometrial cancer using the TCGA dataset. In TCGA-UCEC patients, the overexpression of \u003cem\u003ePDK3\u003c/em\u003e indicated a poor survival outcome. However, \u003cem\u003ePDK3\u003c/em\u003e overexpression contributed to better overall survival in the TCGA-TCGA-BRCA and TCGA-TCGA-CESC datasets.\u003c/p\u003e \u003cp\u003eImmune cells and the tumor microenvironment are decisive in determining tumor progression and metastasis [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. Tumor-infiltrating immune cells have been reported to influence patient survival. Tumor-associated macrophages and neutrophils exhibit either protumorigenic (M2 and N2) or antitumorigenic (M1 and N1) effects on the basis of their polarization status [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Natural killer (NK) cells cause potent cytolysis and antitumour activity in cancer cells [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. Similarly, CD8\u0026thinsp;+\u0026thinsp;and CD4\u0026thinsp;+\u0026thinsp;T cells exhibit increased antitumour activity through granular exocytosis [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. We analysed the link between \u003cem\u003ePDK3\u003c/em\u003e expression and immune cell abundance via TCGA datasets. In the TCGA-BRCA cohort, significantly lower numbers of NK cells and CD8\u0026thinsp;+\u0026thinsp;T cells were observed, which might be responsible for the poor survival outcome. Similarly, in TCGA-UCEC patients, the reduced survival can also be attributed to the significant downregulation of CD8\u0026thinsp;+\u0026thinsp;T cells. The TCGA-CESC dataset revealed significantly increased numbers of unpolarized macrophages and significant downregulation of NK cells. These preliminary observations on tumor infiltration and its involvement in tumor prognosis upon \u003cem\u003ePDK3\u003c/em\u003e expression need to be explored in detail. Previous studies reported the role of \u003cem\u003ePDK3\u003c/em\u003e in therapy resistance, such as in colon cancer [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], gastric cancer [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], and melanoma [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. Many drugs, such as doxorubicin, etoposide, and COL-3, are negatively correlated with \u003cem\u003ePDK3\u003c/em\u003e expression. More detailed studies are needed to understand the role of \u003cem\u003ePDK3\u003c/em\u003e in the development of therapeutic resistance to these drugs.\u003c/p\u003e \u003cp\u003eFinally, we investigated the biological functions of \u003cem\u003ePDK3\u003c/em\u003e in SiHa cells. The knockdown of \u003cem\u003ePDK3\u003c/em\u003e in SiHa cells inhibited both the proliferation and growth of SiHa cells. Our study revealed that \u003cem\u003ePDK3\u003c/em\u003e may promote tumor growth by increasing cell proliferation by decreasing the cell doubling time. Our findings are consistent with previous reports in other cancers, such as urothelial carcinoma [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] and colon cancer [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Overall, we report a procarcinogenic function of PDK3 in gynecological cancer.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eOur study investigated the role of \u003cem\u003ePDK3\u003c/em\u003e in gynecological cancers through extensive computational analysis followed by experimental validation. The gene expression data from multiple databases consistently revealed its overexpression, which emphasizes its role as a potential oncogene. Furthermore, genomic and epigenomic changes have been shown to influence \u003cem\u003ePDK3\u003c/em\u003e expression. \u003cem\u003ePDK3\u003c/em\u003e expression may influence immune cell abundance and clinical outcomes in breast, uterine, endometrial and cervical cancers. Our study identified potential drugs that can be used to treat cancers that overexpress \u003cem\u003ePDK3\u003c/em\u003e. The results of the computational analysis followed by experimental validation suggest an oncogenic role for \u003cem\u003ePDK3\u003c/em\u003e in cervical cancer. These findings clarify the role of \u003cem\u003ePDK3\u003c/em\u003e as a promising biomarker in gynecological cancers.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Wellcome Trust DBT India Alliance, Government of India \u003cstrong\u003e(Grant no: IA/I/22/1/506240)\u003c/strong\u003e\u003cstrong\u003e,\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eawarded to Dr. Shama Prasada K, and support from Joint CSIR-UGC NET Junior Research Fellowship, Government of India \u003cstrong\u003e(Reference ID 221610058423),\u0026nbsp;\u003c/strong\u003eand Dr.\u0026nbsp;TMA Pai structured\u0026nbsp;Ph.D.\u0026nbsp;fellowship.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDr. Shama Prasada Kabekkodu: Conceptualization, Project administration, Resources, Supervision, Visualization, Writing \u0026ndash; review and editing. Medha Bairy: Formal analysis, Investigation, Software, Validation, Writing \u0026ndash; original draft, Visualization. Aishath Shaheeda: Formal analysis, Investigation, Software, Validation, Writing \u0026ndash; original draft, Visualization. Padmanaban S. Suresh: Writing \u0026ndash; review and editing. Roshan Mascarenhas: Writing \u0026ndash; review and editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no ethical statements to disclose since the research does not include any animal or human studies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that all other data supporting the findings of this study are available within the article and its supplementary information files.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eH. Sung, J. Ferlay, R.L. Siegel, M. 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This study was designed to examine the role of \u003cem\u003ePDK3\u003c/em\u003e in gynecological cancers via \u003cem\u003ein silico\u003c/em\u003e analysis and its conformation via \u003cem\u003ein vitro\u003c/em\u003e experiments. Analysis of gynecological cancer datasets from the TCGA and CCLE databases suggested significant upregulation of \u003cem\u003ePDK3\u003c/em\u003e in breast, cervical, and endometrial cancer samples compared with the corresponding normal samples. KEGG pathway enrichment analysis revealed that pathways associated with cancers, viral carcinogenesis, and human papilloma virus infection were highly enriched. \u003cem\u003ePDK3\u003c/em\u003e upregulation affected the overall survival of breast and cervical cancer patients according to Kaplan\u0026ndash;Meier survival analysis. \u003cem\u003ePDK3\u003c/em\u003e upregulation affected the proportion and distribution of immune cells in breast, cervical, and endometrial cancer. Furthermore, we experimentally showed that \u003cem\u003ePDK3\u003c/em\u003e knockdown inhibits the proliferation and growth of SiHa cells. Overall, we found that \u003cem\u003ePDK3\u003c/em\u003e may be an oncogene, and measuring its expression may be of prognostic and therapeutic significance in gynecological cancers.\u003c/p\u003e","manuscriptTitle":"Understanding the Role of Pyruvate Dehydrogenase Kinase 3 in Gynecological Cancers","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-16 13:03:37","doi":"10.21203/rs.3.rs-6429535/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"80e69a55-12df-4591-8357-d78314593667","owner":[],"postedDate":"May 16th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-05-16T13:11:40+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-16 13:03:37","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6429535","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6429535","identity":"rs-6429535","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}