The 3’-UTR Polymorphisms in the NLRP3 Gene Associated with the Risk of COPD and Their Putative Effects on the microRNA Mechanism

The 3’-UTR Polymorphisms in the NLRP3 Gene Associated with the Risk of COPD and Their Putative Effects on the microRNA Mechanism | 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 The 3’-UTR Polymorphisms in the NLRP3 Gene Associated with the Risk of COPD and Their Putative Effects on the microRNA Mechanism Wu Hui Yan, Huang Chu Ting, Zhang Yan Ling, Peng Liang, Li Wei Peng This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3605541/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 Objective Evaluating the association between a single nucleotide polymorphism in the 3'UTR region of the miRNAs binding site of the NLRP3 gene and the occurrence and development of Chronic Obstructive Pulmonary Disease (COPD) and providing information to aid in the early detection and treatment of COPD. Methods The regulatory SNPs located in NLRP3 3'UTR region were searched by using dbSNP database and miRNA binding site prediction database. Meanwhile, samples from COPD patients and healthy controls in the same period were used for verification. The clinical baseline information of all subjects was collected, and the transcription level and protein expression level of NLRP3 and the expression level of inflammatory factors downstream of NLRP3 were detected. The effects of SNPs single nucleotide changes on the transcription and expression of inflammatory factors were analyzed. Results The study included 418 participants (249 in the COPD group and 169 in the control group). NLRP3 SNPs with miRNA binding sites include rs10754558 (G>C), rs1664774076 (ATAT>-), and rs1664775106 (C>G). Furthermore, two genotypes, GCG and GCA, were discovered to have a linkage mutation at 3'UTR 459-461. COPD susceptibility is tightly associated to the expression of the rs1664774076 -/- genotype, and the risk of COPD increased by 3.4 times (P≤0.0001). Type 459-461 GCA was substantially related with the likelihood of developing COPD at various stages (P<0.05). Except for rs10754558, all homozygous mutants increased NLRP3 mRNA and protein levels. NLRP3 had the greatest area under the ROC curve for predicting the development and diagnosis of COPD when compared to its downstream inflammatory variables (AUC= 0.9291). Conclusions The NLRP3 rs1664774076 -/- genotype is a COPD susceptibility gene, and the GCA genotype at 459-461 can be used as an early predictor of COPD exacerbation. The NLRP3 3'UTR polymorphism may alter the loss of miRNA binding sites, leading to an increase in NLRP3 expression. In the development of COPD, NLRP3 has a better diagnostic value than traditional inflammatory factors. Chronic Obstructive Pulmonary Disease NLRP3 3'UTR SNP miRNA Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. Introduction COPD (Chronic Obstructive Pulmonary Disease) is one of the diseases with the highest morbidity and mortality rates worldwide, and the global COPD prevalence rate is on the rise, reaching 9–10% among persons over the age of 40 [ 1 ]. COPD is a lung disease characterized by incomplete reversibility and continuous progressive airflow limitation, with chronic inflammation and structural deterioration of the airway wall and lung parenchyma as its pathological basis. Currently, it has been established that the cause of COPD is the inflammatory reaction of lung tissue, and that long-term stimulation of lung tissue by smoke and pollution is the inducer [ 2 ]. Despite the fact that smoking is a major cause of COPD, between 25 and 45 percent of COPD patients have never smoked, and these patients have a tendency toward regional and familial clustering, suggesting that the onset of COPD is heritable [ 3 ]. Genetic variation analysis may be useful for screening the COPD-vulnerable population. In terms of etiology, the inflammatory response plays a crucial indirect or direct role in the process of lung injury, which is a major cause of decreased lung tissue elasticity, retraction disorder, excessive alveolar expansion, and progressive fibrosis. Mutation of inflammatory factors may result in alterations to the inflammatory response. Therefore, we attempt to construct the inflammatory response based on the molecular genetic mechanism and investigate its relationship with the occurrence and progression of COPD. NLRP 3 (NOD-like Receptor Thermal Protein Domain Associated Protein 3) regulates the occurrence and development of chronic inflammation in COPD as a key factor of inflammatory corpuscles [ 4 ].Faner et al described that NLRP3 inflammatory bodies lead to focal death and passive release of pro-inflammatory cytokines, including mature IL-1β and IL-18, which trigger inflammatory response in COPD patients, and that NLRP3 inflammatory bodies are over-expressed during the deterioration period, and IL-1β and IL-18 family cytokines are released into the surrounding tissues [ 5 ]. Inhibiting the production of NLRP3 inflammatory corpuscles can therefore indirectly block the inflammatory effects of IL-1β and IL-18, making it an ideal therapeutic target for COPD. Gene mutations in the core domain of the NLRP3 gene play a crucial role in the regulation of the immune inflammatory response, resulting in genetic susceptibility to a variety of acute and chronic inflammatory diseases [ 6 ]. In contemporary molecular epidemiology, SNP (Single Nuclear Polymorphism) is frequently used as a biomarker of disease susceptibility. One of the mechanisms by which SNPs affect disease susceptibility is by modulating the interaction between miRNA and mRNA to modulate gene expression. MiRNA(MicroRNAs) can precisely bind to the 3'UTR sequence of the target gene mRNA to inhibit translation or promote degradation of the target gene at the post-transcriptional level. Therefore, this study hypothesizes that the genetic variation of the NLRP3 3'UTR region influences the binding of miRNA-mRNA, resulting in a change in the NLRP3 protein level and thereby influencing the occurrence or development of COPD. In this study, we used the prediction website of miRNA target genes and their binding sites to identify meaningful 3'UTR NLRP3 SNP, and investigated the relationship between these SNP and COPD susceptibility or progression in order to provide a new perspective for the exploration of its pathogenesis or potential therapeutic targets from the research of COPD genomics, and to identify genetic risk factors that can contribute to the early detection, early diagnosis, and prevention of COPD. 2. Material and methods Study Population All of the subjects are from the Fifth Affiliated Hospital of GuangZhou Medical University and have received approval from the institution's Ethics Review Committee. Inclusion and exclusion criteria were as follows: COPD was chosen based on diagnostic guidelines for COPD [7]. Exclude patients with additional chronic respiratory conditions, including lung cancer, thyroid cancer, and other malignant tumors. Have significant immune deficiency and immune system diseases. Antibiotics were administered within 30 days prior to selection. Local or systemic infection, etc. In addition, according to the GOLD(Global Initiative for Chronic Obstructive Lung Disease) standard, the COPD cohort patients were divided into the following categories: GOLD1, mild patients: FEV1%pred≥70%. GOLD2 , moderate patients: 50%≤FEV1%pred<70%. GOLD3, severe patients: 30%≤FEV1%pred<50%. GOLD4, extremely severe patients: FEV1%pred<30%. Standard of healthy control group: healthy people from physical examination center in the same period. Screening of miRNA targeting 3′UTR of NLRP3 gene SNPs The sequence of 3'UTR region in NLRP3 was obtained by using NCBI database , and the Nucleic acid sequence was NG_007509.2, which was a gene fragment of NLRP3. All SNP sites of NG_007509.2 were screened from NCBI SNP, that is, all SNPs of NLRP3 3'UTR. MiRNA binding sites of NLRP3 were obtained by using miRNA prediction websites miRDB, miRTarBase, DIANA, miRcode and TargetScan. Under bioinformatics simulation, the sequence of miRNA was aligned with the position of the 3'UTR SNP site, and the SNPs site with the ability to regulate gene expression was identified. Amplifification and Sequencing of 3′UTR of NLRP3 Regions All participants in this study underwent NLRP3 3'UTR gene sequencing. After 12 hours of fasting, 2 mL of EDTA anticoagulant was extracted the following morning, followed by the extraction of whole blood DNA (TIANGEN blood DNA extraction Kit). The target fragment was amplified using PCR (TIANGEN 2 Taq PCR Master Mix II PCR Kit, primer shown in Table 1), and the sequence of the NLRP3 3'UTR was determined using the Sanger sequencing technique. ClustalX is used to compare the mutation situation of the population, and sites with a mutation rate greater than 30%, which are in agreement with the predicted SNPs sites with regulatory function, are selected for inclusion in the study. TABLE 1. Primer sequences for PCR and qRT-PCR Primer name Sequence 5’ →3’ Tm(℃) NLRP3 3′UTR F TCAATTATGAGACAAAAAGTGCGTT 52.2 NLRP3 3′UTR R CAAACCTTTCCCTCCACGAT 53.8 NLRP3 mRNA F CCCCGTGAGTCCCATTA 52.3 NLRP3 mRNA R GACGCCCAGTCCAACAT 52.3 β-actin mRNA F GAGCTACGAGCTGCCTGACG 59.9 β-actin mRNA R GTAGTTTCGTGGATGCCACAG 56.1 Detection of mRNA expression and downstream factor level After fasting for 12 h, all subjects took 2 mL of EDTA anticoagulant and 5 mL of EDTA procoagulant in the morning of the next day. EDTA anticoagulation is used to extract total RNA from whole blood (Total RNA kit of whole blood, HangZhou Xinjing biochemical reagent Development Co., Ltd.). The mRNA was reverse transcribed into cDNA (TIANGEN gDNA Dispelling RT SuperMix cDNA kit), and then the level of NLRP3 mRNA (TIANGEN Talent qPCR kit, & CobasZ480) was detected by qRT-PCR, and the β-actin mRNA was used as the reference gene for calibration (Table 1 for primers). The serum obtained by procoagulant centrifugation is used to detect inflammatory factors, including: IL-1β(ELISA, Shanghai Jihe Biotechnology Co., Ltd.), IL-18(ELISA, Shanghai Jihe Biotechnology Co., Ltd.), NLRP3(ELISA, Shanghai Jihe Biotechnology Co., Ltd.) and IL-6(ECLIA, ROCHE). Statistical Analysis Statistical examination SPSS27.0 was utilized for statistical analysis of the collected data, while GraphPad Prism 8.0 was utilized for diagramming. C 2 test was used to examine the difference in genotype and allele frequency between groups. C 2 test was used to determine whether the genotype distribution conforms to the Hardy-Weinberg equilibrium law, and sample representativeness was confirmed. The odds ratio (OR) and 95% confidence interval (CI) of logistic regression analysis were used to estimate the relationship between NLRP3 gene polymorphism and the likelihood of COPD occurrence and development. The measured data conforming to the normal distribution are expressed as `X±SD, and T test is used between two groups, and One-way ANOVA test is used for comparison among multiple groups.The measured data that do not conform to the normal distribution are expressed as M(Q1,Q3), and the M-W U test and the Welch ANOVA test are used to compare two groups. For correlation analysis, Pearson correlation analysis is used. P <0.05 is statistically significant. Receiver operating characteristic curve analysis was used to predict the value, and the Area Under the Curve (AUC) and confidence interval were obtained. 3. Results Population Characteristics From August 2021 to March 2023, a total of 168 healthy controls and 221 COPD patients participated. Compared to the control group, the proportion of men in the total COPD group is greater ( P = 0.0039), as is the degree of aging ( P ＜ 0.0001), but the height and weight have no effect on the incidence of COPD (BMI, P= 0.33). To exclude lung function-related diseases from the control group of healthy individuals, this study did not acquire the corresponding lung function data for this group. There is no difference in age, BMI index, or smoking as COPD progresses from mild to severe (P = 0.583, 0.141, and 0.272, respectively). Nevertheless, pulmonary function deteriorated significantly ( P ＜ 0.001), and FVC, FVCpred%, FEV1 and FEVpred% decreased progressively based on subgroup classification. As a result of pulmonary ventilation dysfunction, PaO 2 fell and PaCO 2 rose proportionally (table 2). TABLE 2. Baseline characteristics of control subjects, and Each subgroup of COPD patients. Characteristics GOLD 1 ( n = 74) GOLD 2 ( n =62 ) GOLD3-4 ( n = 85) P values Total COPD ( n =221 ) Controls ( n =168 ) P values Sex, male ( n , %) 58(78.38) 52(83.87) 79(92.94) 0.031 189(85.52 ) 124(73.80) 0.0039 Age ,year 75.42±8.37 77.00±10.24 76.60±9.59 0.583 78(70.00,83.00) 57(50.00,68.00) ＜0.0001 BMI, Kg/m 2 23.48±3.06 20.76±3.18 21.99±3.20 0.141 21.8(20.15,24.39) 21.33(20.26,22.31) 0.3334 Smoking status Never smoker(n, %) 28(37.84) 32(51.61) 38(44.71) 0.272 98(%) - - Lung function FVC, L 3.17±0.79 2.28±0.54 1.81±0.39 ＜0.0001 2.40±0.83 - - FVCpred% 97.15±14.90 80.87±7.73 54.84±21.48 ＜0.0001 76.28±21.48 - - FEV 1 , L 2.29±0.77 1.24±0.29 0.91±0.78 ＜0.0001 1.46±0.78 - - FEV 1 pred% 91.46±14.12 59.01±4.69 36.51±7.45 ＜0.0001 61.17±25.50 - - FEV 1 /FVC% 71.48±8.91 54.70±6.86 51.71±12.38 ＜0.0001 59.14±13.30 - - PaO2, mmHg 96.86±31.90 87.86±21.34 73.41±15.05 0.0008 85.52±25.20 - - PaCO2, mmHg 43.31±8.51 45.32±10.45 54.68±14.30 0.0003 47.95±12.45 - - Abbreviations: -, No data was collected.GOLD, the Global Initiative for Chronic Lung Disease; BMI, Body Mass Index; FVC, Forced vital capacity; FVCpredicted% , the ratio of FVC to its predicted value; FEV1, Forced expiratory volume in the first second; FEV1predicted%, the ratio of FEV1 to its predicted value; FEV1/FVC, the ratio of FEV1 to FVC; PaO2, arterial partial pressure of oxygen ; PaCO2, arterial partial pressure of Carbon Dioxide. NLRP3 3'UTR SNPs verified by the subject samples To cover as many SNPs as feasible in the 3'UTR region, we determined the sequence information of transcripts in the 3'UTR region and then determined the corresponding sequence position of transcripts from NCBI. 86 SNPs out of 256 can bind to miRNA. As population verification, we used the NLRP3 3'UTR gene sequences of all the subjects included in this analysis, and ultimately, six SNPs met the inclusion criteria (Fig.1&Table 3). One miRNA can modulate dozens of genes, and multiple mirnas can recognize the same mRNA target. If there are multiple regulatory relationships, the effect on downstream expression will be greater. rs10754558(G/C) is recognized by miR-3529-3p and miR-549a, and rs1664774076 (ATAT>-) is recognized by miR-5011-5p and miR-1277-5p, according to the findings of this study. According to the results of five miRNA binding site prediction websites, miR-22-3p and miR-223-3p were highly predicted, and rs1664775106 (C>G) and rs200307362 (A>G) were recognized by miR-22-3p and miR-223-3p, respectively. The species with the strongest binding strength is 8mer, whose recognition sites are rs1664774076(ATAT>-) and rs1664775106(C>G) (Table 2). In this investigation, the population distribution of the research object at the locus rs1664775106(C>G) revealed a continuous mutation in the miR-22-3p binding seed zone, designated 459-461(CAG>GCG/GCA). The 3bp-long mutation is located between the 457th and 464th sites in the NLRP3 3'UTR, and the SNP site of RS 1664775106 (C > G) is at the 457th site (Fig.2). Association between NLRP3 3’UTR Polymorphisms and COPD Prevalence In this study, the gene frequencies of rs10754558 (G>C), rs1664774076 (ATAT>-) and rs1664775106 (C>G) all conform to Hardy-Weinberg genetic balance law and are representative of the population ( P >0.05, Table 4). However, in the study population, there were no homozygous mutation phenotypes in rs1445597018(G/T), rs1664772078(A>C) and rs200307362(A>G), and Hardy-Weinberg statistics were P T), rs1664772078(A>C) and rs200307362 (A > G) were not involved in the follow-up study and analysis. The frequencies of polymorphism of rs1664774076 (ATAT>-) and rs1664775106(C>G) were significantly different between COPD patients and control group. Polymorphisms of rs1664774076 (ATAT>-) and rs1664775106 (C>G) are associated with the occurrence of COPD C>G (rs1664774076 ATAT/- vs ATAT/ATAT：OR= 2.221，95%CI = 1.270~3.885， P = 0.005. -/- vs ATAT/ATAT：OR =3.400， 95% CI = 1.952~5.923， P ＜ 0.0001. Allele - vs Allele ATAT：OR=2.035，95% CI = 1.519~2.728， P ＜ 0.0001. rs1664775106 CG vs CC：OR=1.852，95%CI = 1.220~2.811， P =0.004.GG vs CC：OR = 3.409， 95% CI = 1.200~9.680， P = 0.021. Allele G vs Allele C：OR = 1.699，95% CI = 1.228~2.351， P = 0.001）. However, only C allele at rs10754558 and GCA genotype at 459-461 were associated with the risk of COPD (rs10754558 Allele C vs Allele G: OR = 2.638, 95% CI=1.960~3.550, P < 0.0001). 459-461 GCA vs CAG: OR = 2.381, 95% CI=1.452~3.904, P =0.001) (Table 4). TABLE 4. Genotype frequencies of NLRP3 3UTR gene polymorphisms in control subjects and COPD patients. Variant Controls （ n = 168 ） Total COPD （ n =221 ） OR （95% CI） P values rs10754558 （G>C） GG 16（9.52） 35（15.84） 1.000 （reference） GC 73（43.45） 131（59.28） 0.820（0.425~1.583） 0.555 CC 79（47.02） 55（24.89） 3.142（1.585~6.229） 0.001 Allele G 105（31.25） 201（45.48） 1.000 （reference） Allele C 231（68.75） 241（54.52） 2.638（1.960~3.550） ＜0.0001 HWE P 0.998 0.191 rs1664774076 （ATAT> -） ATAT/ATAT 51（30.36） 30（13.57） 1.000 （reference） ATAT/- 62（36.90） 81（36.65） 2.221（1.270~3.885） 0.005 -/- 55（32.74） 110（49.77） 3.400（1.952~5.923） ＜0.0001 Allele ATAT 164（48.81） 141（31.90） 1.000 （reference） Allele - 172（51.19） 301（68.10） 2.035（1.519~2.728） ＜0.0001 HWE P 0.053 0.246 rs1664775106 （C>G） CC 98（58.33） 92（41.63） 1.000 （reference） CG 65（38.69） 113（51.13） 1.852（1.220~2.811） 0.004 GG 5（2.98） 16（7.24） 3.409（1.200~9.680） 0.021 Allele C 261（77.68） 297（67.19） 1.000 （reference） Allele G 75（22.32） 145（32.81） 1.699（1.228~2.351） 0.001 HWE P 0.568 0.207 459-461（CAG>GCG/GCA） CAG 98（58.33） 92（41.63） 1.000 （reference） GCG 36（21.43） 53（23.98） 1.568（0.942~2.612） 0.084 GCA 34（20.24） 76（34.39） 2.381（1.452~3.904） 0.001 In addition, this investigation investigates the difference in allele frequency between COPD subgroups. In severe COPD, the frequency of the C allele in rs10754558 increased. (GOLD1 vs GOLD3-4: P < 0.05, Fig.3A). The difference of Allele G distribution in rs1664774076 and rs1664775106 is significant in COPD mild to moderate or mild to severe, but there is no difference in allele distribution between moderate and severe (GOLD1 vs GOLD2: P ＜ 0.001 ， GOLD1 vs GOLD3-4： P ＜ 0.001，GOLD2 vs GOLD3-4： P > 0.05, Fig.3B&3C). The frequency of double mutation GCA and triple mutation GCG genotypes in 459-461 loci increased with the severity of COPD (GOLD1 vs GOLD2: P ＜0.001，GOLD1 vs GOLD3-4： P < 0.001, FIG.2D), and at the same time, the distribution difference between moderate and severe tends to weaken (GOLD2 vs GOLD3-4: P < 0.05, Fig.3D). The GCG mutations of rs10754558 Allele C, rs1664774076 Allele -, rs1664775106 Allele G, and 459-461 have a low prediction of moderate to severe COPD development risk, and the degree of risk aggravation is insignificant (GOLD2 vs GOLD3-4 P >0.05). However, the risk of aggravation of mild COPD patients was significantly predicted (GOLD1 vs GOLD2 P ＜0.0001. GOLD1 vs GOLD3-4 P ＜0.0001), except for rs10754558allec (GOLD1 vs GOLD2 P = 0.022. GOLD1 vs GOLD3-4 P =0.04. In particular, the exposure risk of 459-461 GCA continuous triple mutation was statistically significant among COPD patients of varying severity(GOLD1 vs GOLD2 P ＜ 0.0001. GOLD2 vs GOLD3-4 P = 0.012. GOLD1 vs GOLD3-4 P ＜0.0001) (Table 5). TABLE 5. NLRP3 3’UTR SNPs increase the risk of COPD exacerbation Variant GOLD1 VS GOLD 2 OR（95% CI） P values GOLD2 VS GOLD3-4 OR（95% CI） P values GOLD1 VS GOLD3-4 OR（95% CI） P values rs10754558 （G/C） Allele G 1.000 （reference） 1.000（reference） Allele C 1.759（1.084~2.853） 0.022 0.908（0.571~1.443） 0.682 1.596（1.021~2.497） 0.040 rs1664774076 （ATAT> -） Allele ATAT 1.000 （reference） 1.000 （reference） Allele - 3.529（2.100~5.933） ＜0.0001 1.556（0.883~2.741） 0.126 5.4903.297~9.142） ＜0.0001 rs1664775106 （C>G） Allele C 1.000 （reference） 1.000 （reference） Allele G 4.396（2.506~7.710） ＜0.0001 0.710（0.444~1.136） 0.153 3.119（1.825~5.332） ＜0.0001 459-461（CAG>GCG/GCA） CAG 1.000 （reference） 1.000 （reference） GCG 31.667（8.776~114.261） ＜0.0001 0.502（0.194~1.296） 0.154 15.894（5.065~49.880） ＜0.0001 GCA 16.077（6.206~41.650） ＜0.0001 0.315（0.127~0.778） 0.012 5.059（2.275~11.250） ＜0.0001 Difference of Plasma inflammatory Marker Level by Genotypes of COPD patiants The direct downstream factors of NLRP3 are IL-1β and IL-18, while the main activating factor of IL-6 expression is IL-1β. Therefore, we choose to detect the protein levels of NLRP3 and its downstream IL-1β, IL-18 and IL-6 to evaluate the influence of gene changes on transcription and expression. Considering that inducers or exposure factors were required to stimulate the expression of NLRP3 and other proteins, and that these inflammatory factors were all in a low expression state in the control group (Fig.4A), this study investigated whether SNP differences in COPD patients affected the expression of NLRP3 mRNA and downstream factors. The plasma NLRP3 expression level was higher in the double mutant allele groups rs1664774076 and rs1664775106. At sites 459-461, the plasma NLRP3 expression level increased as the number of mutant bases increased. NLPR3 protein expression was substantially different between the 459-461 GCA and rs1664774076 Allele- groups ( P <0.001). However, the locus of rs10754558 is opposite, and the expression level of plasma NLRP3 in the double mutant allele (CC) group is lower than that in the GG+GC group. The expression levels of inflammatory factors IL-1β, IL-18 and IL-6 downstream of NLRP-3 were basically the same among the groups, but we found that the above inflammatory factors were all elevated in GCA group at 459-461 (NLRP3：GCA vs CAG， P ＜0.001. IL-1β：GCA vs CAG， P ＜0.001.IL-18：GCA vs CAG， P ＜0.05 . IL-6：GCA vs CAG， P ＜0.001), as shown in Fig.4B. In order to further study whether there are differences in the expression of NLRP3 mRNA in different SNPs, 50 samples including different genotypes in this study were selected for analysis. The transcription level of NLRP3 in each SNPs group is consistent with the expression of NLRP3 protein (Fig.4C). NLRP3 3'UTR mutant genotype affects transcription regulation and is related to lung function. The relative expression of NLRP3 mRNA was negatively correlated with FEV1/FVC%, FEV1%pred, Pa02/PaCO2 (r= -0.7738, -0.6539, -0.4329, P <0.05). As NLRP3 mRNA levels increased, lung function decreased. Fig.5 demonstrates that FEV1/FVC% has the strongest correlation with the relative expression of mRNA (r = -0.7738), whereas NLRP3 mRNA expression has a moderate correlation with Pa02/PaCO2 (r = -0.4329). From Fig.4C, we know that the relative expression of NLRP3 3'UTR mutant NLRP3 mRNA has increased significantly (rs1664774076 Allele-vs Allele ATAT: P <0.001. 459-461 GCA vs CAG: P <0.001), so we analyzed whether the mutant was related to the decline of lung function. rs1664774076 double deletion genotype (-/-) and GCA genotype at 459-461 were negatively correlated with FEV1/FVC% (r= -0.6220, -0.6235, P < 0.001, Fig.6). The predictive value NLRP3 3’UTR SNPs for the occurrence and development of COPD The diagnostic value of rs1664774076 -/- genotype and GCA genotype at 459-461 locus in predicting the risk of COPD alone or in combination is low (AUC is 0.5852, 0.5708 and 0.5972 respectively). Rs1664774076 -/- genotype is more valuable in predicting the development of COPD than 459-461 GCA genotype (rs1664774076 -/-AUC= 0.7390. 459-461 GCA type AUC= 0.5886), NLRP3 inflammatory factor protein has higher predictive value than its downstream factors IL-6, IL-18 and IL-1β (NLRP3 inflammatory factor protein AUC= 0.9291). The AUC of IL-6, IL-18 and IL-1β were 0.6797, 0.7458 and 0.7841, respectively. The diagnostic value of combining NLRP3 and rs1664774076 -/- was slightly improved (AUC= 0.9301) (Fig.7). 4. Discussion There are signaling molecules that activate inflammasomes in the patient's body, which contribute to the complexity of COPD's pathogenesis. Pathogen-related molecular pattern stimulation due to infection, such as viruses and bacteria [8-9] and damage-related molecular pattern stimulation due to oxidative stress, such as ATP and mitochondrial DNA [10] may be the origins of these signaling molecules.This study confirmed the association between NLRP3 and COPD, and found that the expression of NLRP3 and its downstream inflammatory factors was substantially higher in COPD patients than in healthy controls. This distinction is due to two factors: first, the stimulation of external risk factors. Secondly, the presence of SNPs in the NLRP3 population results in variations in inflammatory expression. This can explain why the mutation of the NLRP3 gene does not cause COPD in healthy individuals, and why the development of COPD patients stimulated by the same external factors is heterogeneous. NLRP3 is located in chromosome region 1q43-q44, with 3 kb upstream of the transcription initiation site (exon and intron) and 2 kb downstream of the termination codon (a total of 37,953 kb) [11]. Recent genetic research has focused on the association between NLRP3 genetic variation and susceptibility to autoimmune and inflammatory diseases [12-15]. However, they all investigate the polymorphism of the exon and intron genes in the NLRP3 coding region. This study concentrates on the 3' untranslated region (3'UTR) and locates the functional site, which has been confirmed by the population, as opposed to remaining at the level of biological prediction. There were continuous mutations at sites 459-461 in the 3'UTR region, and the mutant genotypes were GCG or GCA. rs10754558 CC, rs1664774076 Allele- allele (ATAT deletion), and rs1664775106 are examples. COPD susceptibility is associated with the G allele and the 459-461 GCA genotype, which can enhance the risk of COPD. However, for the demonstration that GG is the susceptible genotype [11] and other articles [16] have reported higher expression of the GG gene NLRP3, one of the possibilities for the difference in results is the regional variation of samples collected, and the effect of the rs10754558 SNP on COPD remains unknown.In addition, the distribution of SNPs differs between subtypes of COPD and is associated with COPD development. Notable is the fact that genotype 459-461 GCA, as a mutant containing three consecutive bases, has the highest degree of mutation, which can not only trigger the early aggravation of risk but also foretell the development risk from GOLD2 to GOLD3-4. Consequently, site mutations are more likely to exacerbate the disease. This study indicates that SNP mutations can increase the likelihood of disease occurrence or disease development. Polymorphisms in the 3'UTR region influence mRNA stability and translation [17], which may substantially affect gene expression by removing, diminishing, or producing miRNA-binding sites. This section of the binding site is also known as the seed region. It consists of 6-7 bases that are complementary to the 3'UTR of the target mRNA. The combination of miRNAs can inhibit the expression of a target protein. The mutants of rs1664774076 (ATAT>-), rs1664775106 (C>G) and 459-461 all showed synchronous up-regulation of NLRP3 mRNA expression and NLRP3 protein level, indicating that there may be a relationship of gene regulation. Different SNP genotypes have no effect on the expression of NLRP3's downstream factors (IL-1β, IL-18, and IL-6). Nevertheless, the 459-461 GCA genotype demonstrated a high level of expression in NLRP3 and its downstream inflammatory factors ( P ＜0.05, Fig.4B), which may indicate that the triple base continuous mutation has a greater regulatory influence on the NLRP3 signaling pathway. The mechanism may be that the 459-461 site is located in the seed region of miR-22-3p, and the mutation of three bases results in the loss of the binding site for miR-22-3p, thereby reducing the inhibitory effect of miR-22-3p on the expression of NLRP3 protein and increasing NLRP3. Previous research has also demonstrated that miR-22-3p inhibits the expression of NLRP3 to reduce the inflammatory effects of gout[18-19]. Rs1664774076 (ATAT>-) is recognized by miR-1277-5p and miR-5011-5p, and the mutation at this site corresponds to the deletion of ATAT. Additionally, there is a loss of four bases in the seed region of miRNA, indicating that the mutation at this site may play a larger role in regulating NLRP3. MiR-1277-5p can bind to the 3'UTR of IRS3(Recombinant Insulin Receptor Substrate 3) mRNA to inhibit its expression and reduce cell damage, according to studies [20]. MiR-223-3p has been demonstrated to be a targeted anti-inflammatory therapeutic molecule, and its down-regulation can reduce inflammation [21-23].Rs200307362(A>G) is located in the miR-223-3p-identified seed zone. In contrast, rs200307362 (A>G) did not contain any double mutant GG in this study. It is inferred that the seed region containing rs200307362 can bind tightly to miR-223-3p, and that miR-223-3p inhibits NLRP3. The level of NLRP3 mRNA increased as lung function declined, and the genotypes of rs1664774076 (ATAT>-) and 459-461 GCA also showed a correlation with lung function decline. The expression levels of NLRP3 mRNA were markedly higher in individuals with the rs1664774076 -/- genotype and the 459-461 GCA genotype. It can be inferred that these two SNP mutations affect the expression of NLRP3, which ultimately reduces COPD patients' lung function and exacerbates the disease. The rs1664774076 (ATAT>-) and 459-461 GCA genotypes can be used for risk prediction, whereas the detection of the NLRP3 gene has a low diagnostic value for COPD and cannot supplant conventional diagnostic methods. COPD acute exacerbation is measured using inflammation indices (such as CRP, IL-6, and IL-1β) [4] .This study discovered that NLRP3 has a greater predictive value than IL-1β and IL-6. In light of the fact that the expression of inflammatory factors has been stimulated by inducing factors, i.e., the expression of inflammatory factors trails behind that of gene detection, it is suggested that the detection of SNPs plus NLRP3 can aid in the clinical evaluation of the progression of a patient's disease. This study has some limitations, to conclude. This analysis is limited to the Chinese population and may not be generalizable to other populations. In addition, the scope of this study is limited to the prediction of the miRNA binding site, and it is still necessary to complete the double luciferase reporter gene verification experiment in order to determine the nature of their relationship. In addition, it is necessary to accomplish the miRNA population survey necessary for this study. If the expression level of the miRNA population is minimal, it will also reduce the regulation of SNPs on inflammatory factors. Declarations Authors’ Contributions: Wu Hui Yan performed miRNAs-bing sites prediction and SNPs screening, data analysis, and article writing. Huang Chu Ting completed the whole experiment and wrote the original draft. Zhang Yan Ling participated in the screening and enrollment of subjects, blood samples collecting, and clinical data collation. Peng Liang contributed to project administration. Li Wei Peng accomplished the funding acquisition and the writing review of the article. All the authors read and approved the final article. Author Disclosure Statement: No competing financial interests exist. Funding: The research was funded by Medical Scientific Research Foundation of Guangdong Province of China (B2021440). Informed Consent Statement: Written informed consent to publish this paper was obtained from the patient(s). Institutional Review Board Statement: The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Review Board (or Ethics Committee) of the Fifth Affiliated Hospital of Guangzhou Medical University (protocol KY01-2020-11-06). Declaration Section: The data that support the SNPs and the sequence of 3'UTR region in NLRP3 of this study are available in NCBI. These miRNA binding sites of NLRP3 were derived from the following resources availiable in the public domain:miRDB, miRTarBase, DIANA, miRcode and TargetScan. 1. NCBI database (The sequence of 3'UTR region in NLRP3 was obtained) accession numbers: Gene ID: 114548 web links: https://www.ncbi.nlm.nih.gov/gene/114548#reference-sequences and the matching NCBI blast to verify accession numbers：NG_007509.2 web links: https://www.ncbi.nlm.nih.gov/projects/sviewer/?id=NG_007509.2&search=NG_007509.2:g.37562G%3EC&v=1:100&content=5 2. dbSNP ( All SNPs of NLRP3 3'UTR were obtained) accession numbers: search key word: NLRP3 web links: https://www.ncbi.nlm.nih.gov/snp/ 3. miRNA prediction websites: miRDB, miRTarBase, DIANA, miRcode and TargetScan 3.1 miRDB accession numbers: search by gene target: NLRP3 web links: https://mirdb.org/mirdb/index.html 3.2 miRTarBase web links: https://mirtarbase.cuhk.edu.cn/~miRTarBase/miRTarBase_2022/php/search.php?org=hsa&kw=NLRP3&opt=target 3.3 DIANA web links: https://dianalab.e-ce.uth.gr/html/dianauniverse/index.php?r=microT_CDS/results&keywords=ENSG00000162711&genes=ENSG00000162711%20&mirnas=&descr=&threshold=0.7 3.4 miRcode web links: http://www.mircode.org/index.php?gene=NLRP3&mirfam=&class=&cons=&trregion= 3.5 TargetScan web links: https://www.targetscan.org/cgi-bin/targetscan/vert_80/view_gene.cgi?rs=ENST00000391828.3&taxid=9606&showcnc=0&shownc=0&shownc_nc=&showncf1=&showncf2=&subset=1 References Vijayan V K. Chronic obstructive pulmonary disease[J]. Indian J Med Res, 2013,137(2):251-269. Vogelmeier C F, Roman-Rodriguez M, Singh D, et al. Goals of COPD treatment: Focus on symptoms and exacerbations[J]. Respir Med, 2020,166:105938. Ortega-Martinez A, Perez-Rubio G, Ambrocio-Ortiz E, et al. The SNP rs13147758 in the HHIP Gene Is Associated With COPD Susceptibility, Serum, and Sputum Protein Levels in Smokers[J]. Front Genet, 2020,11:882. Guo P, Li R, Piao T H, et al. Pathological Mechanism and Targeted Drugs of COPD[J]. Int J Chron Obstruct Pulmon Dis, 2022,17:1565-1575. Chen X, Guo X, Ge Q, et al. ER Stress Activates the NLRP3 Inflammasome: A Novel Mechanism of Atherosclerosis[J]. Oxid Med Cell Longev, 2019,2019:3462530. Lu F, Chen H, Hong Y, et al. A gain-of-function NLRP3 3'-UTR polymorphism causes miR-146a-mediated suppression of NLRP3 expression and confers protection against sepsis progression[J]. Sci Rep, 2021,11(1):13300. Rabe K F, Hurd S, Anzueto A, et al. Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary[J]. Am J Respir Crit Care Med, 2007,176(6):532-555. Martinon F, Agostini L, Meylan E, et al. Identification of bacterial muramyl dipeptide as activator of the NALP3/cryopyrin inflammasome[J]. Curr Biol, 2004,14(21):1929-1934. Kanneganti T D, Ozoren N, Body-Malapel M, et al. Bacterial RNA and small antiviral compounds activate caspase-1 through cryopyrin/Nalp3[J]. Nature, 2006,440(7081):233-236. Riteau N, Gasse P, Fauconnier L, et al. Extracellular ATP is a danger signal activating P2X7 receptor in lung inflammation and fibrosis[J]. Am J Respir Crit Care Med, 2010,182(6):774-783. Xu G, Huang R, Xia W, et al. Associations between inflammasome-related gene NLRP3 Polymorphisms (rs10754558 and rs35829419) and risk of bladder cancer in a Chinese population[J]. J Clin Lab Anal, 2021,35(11):e23973. Wu Z, Wu S, Liang T. Association of NLRP3 rs35829419 and rs10754558 Polymorphisms With Risks of Autoimmune Diseases: A Systematic Review and Meta-Analysis[J]. Front Genet, 2021,12:690860. Hitomi Y, Ebisawa M, Tomikawa M, et al. Associations of functional NLRP3 polymorphisms with susceptibility to food-induced anaphylaxis and aspirin-induced asthma[J]. J Allergy Clin Immunol, 2009,124(4):779-785. Zhao S, Chen H, Wu G, et al. The association of NLRP3 and TNFRSF1A polymorphisms with risk of ankylosing spondylitis and treatment efficacy of etanercept[J]. J Clin Lab Anal, 2017,31(6). Queiroz G A, Da S R, Pires A O, et al. New variants in NLRP3 inflammasome genes increase risk for asthma and Blomia tropicalis-induced allergy in a Brazilian population[J]. Cytokine X, 2020,2(3):100032. Cheng L, Liang X, Qian L, et al. NLRP3 gene polymorphisms and expression in rheumatoid arthritis[J]. Exp Ther Med, 2021,22(4):1110. Kim J O, Park H S, Ko E J, et al. The 3'-UTR Polymorphisms in the Thymidylate Synthase (TS) Gene Associated with the Risk of Ischemic Stroke and Silent Brain Infarction[J]. J Pers Med, 2021,11(3). Wang X, Chi J, Dong B, et al. MiR-223-3p and miR-22-3p inhibit monosodium urate-induced gouty inflammation by targeting NLRP3[J]. Int J Rheum Dis, 2021,24(4):599-607. Zhang Q B, Zhu D, Dai F, et al. MicroRNA-223 Suppresses IL-1beta and TNF-alpha Production in Gouty Inflammation by Targeting the NLRP3 Inflammasome[J]. Front Pharmacol, 2021,12:637415. Sun Z, Song L, Li J. Knockdown of small nucleolar RNA host gene 10 (SNHG10) alleviates the injury of human neuroblastoma cells via the miR-1277-5p/insulin substrate receptor 2 axis[J]. Bioengineered, 2022,13(1):709-720. Dong H C, Li P N, Chen C J, et al. Sinomenine Attenuates Cartilage Degeneration by Regulating miR-223-3p/NLRP3 Inflammasome Signaling[J]. Inflammation, 2019,42(4):1265-1275. Zhao G, Jiang K, Yang Y, et al. The Potential Therapeutic Role of miR-223 in Bovine Endometritis by Targeting the NLRP3 Inflammasome[J]. Front Immunol, 2018,9:1916. Neudecker V, Haneklaus M, Jensen O, et al. Myeloid-derived miR-223 regulates intestinal inflammation via repression of the NLRP3 inflammasome[J]. J Exp Med, 2017,214(6):1737-1752. Additional Declarations No competing interests reported. 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. 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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-3605541","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":252330906,"identity":"38692fce-16a4-4cf3-991d-e9b05509832a","order_by":0,"name":"Wu Hui Yan","email":"","orcid":"","institution":"The Fifth Affiliated Hospital of Guangzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Wu","middleName":"Hui","lastName":"Yan","suffix":""},{"id":252330908,"identity":"d9eec415-61ce-4b05-a67e-8107ac9349f1","order_by":1,"name":"Huang Chu Ting","email":"","orcid":"","institution":"KingMed School of Laboratory Medicine of Guangzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Huang","middleName":"Chu","lastName":"Ting","suffix":""},{"id":252330910,"identity":"8bb07b0a-1cd0-49fb-8df9-45c89b081cb8","order_by":2,"name":"Zhang Yan Ling","email":"","orcid":"","institution":"The Fifth Affiliated Hospital of Guangzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zhang","middleName":"Yan","lastName":"Ling","suffix":""},{"id":252330912,"identity":"0580b6e7-4223-4e92-ad0c-47ea1cd6944d","order_by":3,"name":"Peng Liang","email":"","orcid":"","institution":"The Fifth Affiliated Hospital of Guangzhou Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Peng","middleName":"","lastName":"Liang","suffix":""},{"id":252330916,"identity":"22d16f16-d6b0-4184-b5ff-298695436b5e","order_by":4,"name":"Li Wei Peng","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIie3PQUrDQBSA4RcG4ubpbBsUeoUXCooQqkeZRyErEcELDBRyhnTVK+gmdvlKoauiB3BhvMEENwqCptnU1aRLofMvhpnhfTADEAr9w05Uu5huq0QMZai19ZN4R2Ku3V1+lpTSQ3ZbHKWlW2VkTQ85wrR+X2S3w2s4P0V6QQKJXHPjexiOiDf5fSqQt+QVL5RVyazykeNqwMWKHy2sO3JpZXvpJU+fXPy0JCpa8owkppdUwIXwHJRKS5J9iP4Y8GbCDxBHtaMJJuVy6v2L1mtuvhZjnlvtxHyPr7SeLl3jIV2RBaC3+u+xr+3McI+5UCgUOtB+AdBbT9f3uJVuAAAAAElFTkSuQmCC","orcid":"","institution":"Wuhan Dian Medical Laboratory Co., Ltd","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Li","middleName":"Wei","lastName":"Peng","suffix":""}],"badges":[],"createdAt":"2023-11-13 14:29:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3605541/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3605541/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":47383440,"identity":"0a243a4f-a976-4309-a65a-13c63f16da86","added_by":"auto","created_at":"2023-11-30 17:02:47","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":34666,"visible":true,"origin":"","legend":"\u003cp\u003eScreening of NLRP3 3’UTR SNPs\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/c6a36639af8969f8a4aa1188.jpg"},{"id":47383441,"identity":"57400dc9-5da2-4551-98f5-4a5c6bb1f97d","added_by":"auto","created_at":"2023-11-30 17:02:47","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":21570,"visible":true,"origin":"","legend":"\u003cp\u003eThe continuous mutation is located between the 457th and 464th sites in the NLRP3 3'UTR\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/5f9e2c0386bc17909777097c.jpg"},{"id":47384228,"identity":"afbcbd27-ed88-4615-b5c5-8f98a12524f6","added_by":"auto","created_at":"2023-11-30 17:10:47","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":53456,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution of SNPs alleles among different subgroups of COPD. (ns：no significance；*\u003cem\u003eP\u003c/em\u003e＜0.05；**\u003cem\u003eP\u003c/em\u003e＜0.001)\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/44568fd664954ff07f16f20f.jpg"},{"id":47383445,"identity":"1e605438-4769-43ef-af9c-8b56a684bf83","added_by":"auto","created_at":"2023-11-30 17:02:47","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":126548,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePotential regulatory effects of NLRP3 gene subtypes on downstream expressed proteins\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAbbreviations: A. The difference in the expression of inflammatory factor protein downstream of NLRP3 between the control group and COPD patients. B. The expression of NLRP3 protein and downstream factors in each SNPs. C. The expression of NLRP3 mRNA in each SNPs;（ns：no significance；*P＜0.05； **P＜0.001）\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/215af01ee9f8c90a3fb9555a.jpg"},{"id":47384227,"identity":"e2ea2c30-565f-4956-adcd-75b0151e66b5","added_by":"auto","created_at":"2023-11-30 17:10:47","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":32431,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation analysis between lung function and relative expression of NLRP3 mRNA.\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/6d418da5bd1b1e964d142680.jpg"},{"id":47383442,"identity":"cdd45af3-3023-4b91-8a49-4829af0bfad5","added_by":"auto","created_at":"2023-11-30 17:02:47","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":22801,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation analysis between lung function and NLRP3 3'UTR mutant genotype\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/f6b6356ae9f999bcbe4aaf68.jpg"},{"id":47383443,"identity":"1d65219f-c108-4195-90c0-0e21745797df","added_by":"auto","created_at":"2023-11-30 17:02:47","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":47511,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eROC curve of the combination of plasma inflammatory markers and SNPs\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAbbreviations: A: ROC curve of predicting the risk of COPD by SNPs (positive sample in COPD group and negative sample in control group); B: The ROC curve of NLRP3 polymorphism and inflammatory factor level for predicting the aggravation of COPD development risk (with GOLD3-4 group as positive sample and GOLD1 group as control group)\u003c/p\u003e","description":"","filename":"7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/979c07b26cbf3d8c0ea98457.jpg"},{"id":56595951,"identity":"0f2debb1-fcdf-40c1-a290-397114eb9bc4","added_by":"auto","created_at":"2024-05-16 10:30:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1011595,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3605541/v1/ff129f4f-7d40-47d8-8596-fb3e93d48977.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"The 3’-UTR Polymorphisms in the NLRP3 Gene Associated with the Risk of COPD and Their Putative Effects on the microRNA Mechanism","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCOPD (Chronic Obstructive Pulmonary Disease) is one of the diseases with the highest morbidity and mortality rates worldwide, and the global COPD prevalence rate is on the rise, reaching 9\u0026ndash;10% among persons over the age of 40 [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. COPD is a lung disease characterized by incomplete reversibility and continuous progressive airflow limitation, with chronic inflammation and structural deterioration of the airway wall and lung parenchyma as its pathological basis. Currently, it has been established that the cause of COPD is the inflammatory reaction of lung tissue, and that long-term stimulation of lung tissue by smoke and pollution is the inducer [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Despite the fact that smoking is a major cause of COPD, between 25 and 45 percent of COPD patients have never smoked, and these patients have a tendency toward regional and familial clustering, suggesting that the onset of COPD is heritable [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Genetic variation analysis may be useful for screening the COPD-vulnerable population. In terms of etiology, the inflammatory response plays a crucial indirect or direct role in the process of lung injury, which is a major cause of decreased lung tissue elasticity, retraction disorder, excessive alveolar expansion, and progressive fibrosis. Mutation of inflammatory factors may result in alterations to the inflammatory response. Therefore, we attempt to construct the inflammatory response based on the molecular genetic mechanism and investigate its relationship with the occurrence and progression of COPD.\u003c/p\u003e \u003cp\u003eNLRP 3 (NOD-like Receptor Thermal Protein Domain Associated Protein 3) regulates the occurrence and development of chronic inflammation in COPD as a key factor of inflammatory corpuscles [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].Faner et al described that NLRP3 inflammatory bodies lead to focal death and passive release of pro-inflammatory cytokines, including mature IL-1β and IL-18, which trigger inflammatory response in COPD patients, and that NLRP3 inflammatory bodies are over-expressed during the deterioration period, and IL-1β and IL-18 family cytokines are released into the surrounding tissues [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Inhibiting the production of NLRP3 inflammatory corpuscles can therefore indirectly block the inflammatory effects of IL-1β and IL-18, making it an ideal therapeutic target for COPD.\u003c/p\u003e \u003cp\u003eGene mutations in the core domain of the NLRP3 gene play a crucial role in the regulation of the immune inflammatory response, resulting in genetic susceptibility to a variety of acute and chronic inflammatory diseases [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. In contemporary molecular epidemiology, SNP (Single Nuclear Polymorphism) is frequently used as a biomarker of disease susceptibility. One of the mechanisms by which SNPs affect disease susceptibility is by modulating the interaction between miRNA and mRNA to modulate gene expression. MiRNA(MicroRNAs) can precisely bind to the 3'UTR sequence of the target gene mRNA to inhibit translation or promote degradation of the target gene at the post-transcriptional level. Therefore, this study hypothesizes that the genetic variation of the NLRP3 3'UTR region influences the binding of miRNA-mRNA, resulting in a change in the NLRP3 protein level and thereby influencing the occurrence or development of COPD.\u003c/p\u003e \u003cp\u003eIn this study, we used the prediction website of miRNA target genes and their binding sites to identify meaningful 3'UTR NLRP3 SNP, and investigated the relationship between these SNP and COPD susceptibility or progression in order to provide a new perspective for the exploration of its pathogenesis or potential therapeutic targets from the research of COPD genomics, and to identify genetic risk factors that can contribute to the early detection, early diagnosis, and prevention of COPD.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cp\u003e\u003cem\u003eStudy Population\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAll of the subjects are from the Fifth Affiliated Hospital of GuangZhou Medical University and have received approval from the institution\u0026apos;s Ethics Review Committee. Inclusion and exclusion criteria were as follows:\u003c/p\u003e\n\u003cp\u003eCOPD was chosen based on diagnostic guidelines for COPD [7]. Exclude patients with additional chronic respiratory conditions, including lung cancer, thyroid cancer, and other malignant tumors. Have significant immune deficiency and immune system diseases. Antibiotics were administered within 30 days prior to selection. Local or systemic infection, etc. In addition, according to the GOLD(Global Initiative for Chronic Obstructive Lung Disease) standard, the COPD cohort patients were divided into the following categories: GOLD1, mild patients: FEV1%pred\u0026ge;70%. GOLD2 , moderate patients: 50%\u0026le;FEV1%pred\u0026lt;70%. GOLD3, severe patients: 30%\u0026le;FEV1%pred\u0026lt;50%. GOLD4, extremely severe patients: FEV1%pred\u0026lt;30%. Standard of healthy control group: healthy people from physical examination center in the same period.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;Screening of miRNA targeting 3\u0026prime;UTR of NLRP3 gene SNPs\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe sequence of 3\u0026apos;UTR region in NLRP3 was obtained by using NCBI database , and the Nucleic acid sequence was NG_007509.2, which was a gene fragment of NLRP3. All SNP sites of NG_007509.2 were screened from NCBI SNP, that is, all SNPs of NLRP3 3\u0026apos;UTR. MiRNA binding sites of NLRP3 were obtained by using miRNA prediction websites miRDB, miRTarBase, DIANA, miRcode and TargetScan. Under bioinformatics simulation, the sequence of miRNA was aligned with the position of the 3\u0026apos;UTR SNP site, and the SNPs site with the ability to regulate gene expression was identified.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAmplifification and Sequencing of 3\u0026prime;UTR of NLRP3 Regions\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAll participants in this study underwent NLRP3 3\u0026apos;UTR gene sequencing. After 12 hours of fasting, 2 mL of EDTA anticoagulant was extracted the following morning, followed by the extraction of whole blood DNA (TIANGEN blood DNA extraction Kit). The target fragment was amplified using PCR (TIANGEN 2 Taq PCR Master Mix II PCR Kit, primer shown in Table 1), and the sequence of the NLRP3 3\u0026apos;UTR was determined using the Sanger sequencing technique. ClustalX is used to compare the mutation situation of the population, and sites with a mutation rate greater than 30%, which are in agreement with the predicted SNPs sites with regulatory function, are selected for inclusion in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTABLE 1.\u003c/strong\u003e Primer sequences for PCR and qRT-PCR\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003ePrimer name\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eSequence 5\u0026rsquo; \u0026rarr;3\u0026rsquo;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003eTm(℃)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003eNLRP3\u0026nbsp;3\u0026prime;UTR\u0026nbsp;F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eTCAATTATGAGACAAAAAGTGCGTT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003e52.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003eNLRP3\u0026nbsp;3\u0026prime;UTR\u0026nbsp;R\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eCAAACCTTTCCCTCCACGAT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003e53.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003eNLRP3 mRNA F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eCCCCGTGAGTCCCATTA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003e52.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003eNLRP3 mRNA R\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eGACGCCCAGTCCAACAT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003e52.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026beta;-actin mRNA F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eGAGCTACGAGCTGCCTGACG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003e59.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"23.008849557522122%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026beta;-actin mRNA R\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"48.49557522123894%\" valign=\"top\"\u003e\n \u003cp\u003eGTAGTTTCGTGGATGCCACAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.495575221238937%\" valign=\"top\"\u003e\n \u003cp\u003e56.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cem\u003eDetection of mRNA expression and downstream factor level\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eAfter fasting for 12 h, all subjects took 2 mL of EDTA anticoagulant and 5 mL of EDTA procoagulant in the morning of the next day. EDTA anticoagulation is used to extract total RNA from whole blood (Total RNA kit of whole blood, HangZhou Xinjing biochemical reagent Development Co., Ltd.). The mRNA was reverse transcribed into cDNA (TIANGEN gDNA Dispelling RT SuperMix cDNA kit), and then the level of NLRP3 mRNA (TIANGEN Talent qPCR kit, \u0026amp; CobasZ480) was detected by qRT-PCR, and the \u0026beta;-actin mRNA was used as the reference gene for calibration (Table 1 for primers). The serum obtained by procoagulant centrifugation is used to detect inflammatory factors, including: IL-1\u0026beta;(ELISA, Shanghai Jihe Biotechnology Co., Ltd.), IL-18(ELISA, Shanghai Jihe Biotechnology Co., Ltd.), NLRP3(ELISA, Shanghai Jihe Biotechnology Co., Ltd.) and IL-6(ECLIA, ROCHE).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eStatistical Analysis\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eStatistical examination SPSS27.0 was utilized for statistical analysis of the collected data, while GraphPad Prism 8.0 was utilized for diagramming.\u0026nbsp;C\u003csup\u003e2\u003c/sup\u003e test was used to examine the difference in genotype and allele frequency between groups. C\u003csup\u003e2\u003c/sup\u003e test was used to determine whether the genotype distribution conforms to the Hardy-Weinberg equilibrium law, and sample representativeness was confirmed. The odds ratio (OR) and 95% confidence interval (CI) of logistic regression analysis were used to estimate the relationship between NLRP3 gene polymorphism and the likelihood of COPD occurrence and development. The measured data conforming to the normal distribution are expressed as `X\u0026plusmn;SD, and T test is used between two groups, and One-way ANOVA test is used for comparison among multiple groups.The measured data that do not conform to the normal distribution are expressed as M(Q1,Q3), and the M-W U test and the Welch ANOVA test are used to compare two groups. For correlation analysis, Pearson correlation analysis is used. \u003cem\u003eP\u003c/em\u003e\u0026lt;0.05 is statistically significant. Receiver operating characteristic curve analysis was used to predict the value, and the Area Under the Curve (AUC) and confidence interval were obtained.\u003c/p\u003e"},{"header":"3. Results","content":"\u003cp\u003e\u003cem\u003ePopulation Characteristics\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eFrom August 2021 to March 2023, a total of 168 healthy controls and 221 COPD patients participated. Compared to the control group, the proportion of men in the total COPD group is greater (\u003cem\u003eP\u003c/em\u003e= 0.0039), as is the degree of aging (\u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e0.0001), but the height and weight have no effect on the incidence of COPD (BMI, P= 0.33). To exclude lung function-related diseases from the control group of healthy individuals, this study did not acquire the corresponding lung function data for this group. There is no difference in age, BMI index, or smoking as COPD progresses from mild to severe (P = 0.583, 0.141, and 0.272, respectively). Nevertheless, pulmonary function deteriorated significantly (\u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e0.001), and FVC, FVCpred%, FEV1 and FEVpred% decreased progressively based on subgroup classification. As a result of pulmonary ventilation dysfunction,\u0026nbsp;PaO\u003csub\u003e2\u003c/sub\u003e fell and PaCO\u003csub\u003e2\u003c/sub\u003e rose proportionally (table 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTABLE 2.\u0026nbsp;\u003c/strong\u003eBaseline characteristics of control subjects, and Each subgroup of COPD patients.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"646\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eCharacteristics\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003eGOLD 1\u003c/p\u003e\n \u003cp\u003e(\u003cem\u003en\u0026nbsp;\u003c/em\u003e= 74)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003eGOLD 2\u003c/p\u003e\n \u003cp\u003e(\u003cem\u003en\u0026nbsp;\u003c/em\u003e=62 )\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003eGOLD3-4\u003c/p\u003e\n \u003cp\u003e(\u003cem\u003en\u0026nbsp;\u003c/em\u003e= 85)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalues\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eTotal COPD\u003c/p\u003e\n \u003cp\u003e(\u003cem\u003en\u0026nbsp;\u003c/em\u003e=221 )\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003eControls\u003c/p\u003e\n \u003cp\u003e(\u003cem\u003en\u0026nbsp;\u003c/em\u003e=168 )\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalues\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eSex, male (\u003cem\u003en\u003c/em\u003e, %)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e58(78.38)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e52(83.87)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e79(92.94)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e0.031\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e189(85.52 )\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e124(73.80)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e0.0039\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eAge ,year\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e75.42\u0026plusmn;8.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e77.00\u0026plusmn;10.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e76.60\u0026plusmn;9.59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e0.583\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e78(70.00,83.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e57(50.00,68.00)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eBMI, Kg/m\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e23.48\u0026plusmn;3.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e20.76\u0026plusmn;3.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e21.99\u0026plusmn;3.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e0.141\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e21.8(20.15,24.39)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e21.33(20.26,22.31)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e0.3334\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eSmoking status\u003c/p\u003e\n \u003cp\u003eNever smoker(n, %)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\"\u003e\n \u003cp\u003e28(37.84)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\"\u003e\n \u003cp\u003e32(51.61)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\"\u003e\n \u003cp\u003e38(44.71)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\"\u003e\n \u003cp\u003e0.272\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\"\u003e\n \u003cp\u003e98(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"8\" valign=\"top\"\u003e\n \u003cp\u003eLung function\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eFVC, L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e3.17\u0026plusmn;0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e2.28\u0026plusmn;0.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e1.81\u0026plusmn;0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e2.40\u0026plusmn;0.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eFVCpred%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e97.15\u0026plusmn;14.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e80.87\u0026plusmn;7.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e54.84\u0026plusmn;21.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e76.28\u0026plusmn;21.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eFEV\u003csub\u003e1\u003c/sub\u003e, L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e2.29\u0026plusmn;0.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e1.24\u0026plusmn;0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e0.91\u0026plusmn;0.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e1.46\u0026plusmn;0.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eFEV\u003csub\u003e1\u003c/sub\u003epred%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e91.46\u0026plusmn;14.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e59.01\u0026plusmn;4.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e36.51\u0026plusmn;7.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e61.17\u0026plusmn;25.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003eFEV\u003csub\u003e1\u003c/sub\u003e/FVC%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e71.48\u0026plusmn;8.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e54.70\u0026plusmn;6.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e51.71\u0026plusmn;12.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e59.14\u0026plusmn;13.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003ePaO2, mmHg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e96.86\u0026plusmn;31.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e87.86\u0026plusmn;21.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e73.41\u0026plusmn;15.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e0.0008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e85.52\u0026plusmn;25.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003ePaCO2, mmHg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.145510835913313%\" valign=\"top\"\u003e\n \u003cp\u003e43.31\u0026plusmn;8.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.609907120743035%\" valign=\"top\"\u003e\n \u003cp\u003e45.32\u0026plusmn;10.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.455108359133128%\" valign=\"top\"\u003e\n \u003cp\u003e54.68\u0026plusmn;14.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.13312693498452%\" valign=\"top\"\u003e\n \u003cp\u003e0.0003\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.634674922600619%\" valign=\"top\"\u003e\n \u003cp\u003e47.95\u0026plusmn;12.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.09907120743034%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.287925696594428%\" valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: -, No data was collected.GOLD, the Global Initiative for Chronic Lung Disease; BMI, Body Mass Index; FVC, Forced vital capacity; FVCpredicted% , the ratio of FVC to its predicted value; FEV1, Forced expiratory volume in the first second; FEV1predicted%, the ratio of FEV1 to its predicted value; FEV1/FVC, the ratio of FEV1 to FVC; PaO2, arterial partial pressure of oxygen ; PaCO2, arterial partial pressure of Carbon Dioxide.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eNLRP3 3\u0026apos;UTR SNPs verified by the subject samples\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eTo cover as many SNPs as feasible in the 3\u0026apos;UTR region, we determined the sequence information of transcripts in the 3\u0026apos;UTR region and then determined the corresponding sequence position of transcripts from NCBI. 86 SNPs out of 256 can bind to miRNA. As population verification, we used the NLRP3 3\u0026apos;UTR gene sequences of all the subjects included in this analysis, and ultimately, six SNPs met the inclusion criteria (Fig.1\u0026amp;Table 3).\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\" width=\"679\" height=\"920\"\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eOne miRNA can modulate dozens of genes, and multiple mirnas can recognize the same mRNA target. If there are multiple regulatory relationships, the effect on downstream expression will be greater. rs10754558(G/C) is recognized by miR-3529-3p and miR-549a, and rs1664774076 (ATAT\u0026gt;-) is recognized by miR-5011-5p and miR-1277-5p, according to the findings of this study. According to the results of five miRNA binding site prediction websites, miR-22-3p and miR-223-3p were highly predicted, and rs1664775106 (C\u0026gt;G) and rs200307362 (A\u0026gt;G) were recognized by miR-22-3p and miR-223-3p, respectively. The species with the strongest binding strength is 8mer, whose recognition sites are rs1664774076(ATAT\u0026gt;-) and rs1664775106(C\u0026gt;G) (Table 2). In this investigation, the population distribution of the research object at the locus rs1664775106(C\u0026gt;G) revealed a continuous mutation in the miR-22-3p binding seed zone, designated 459-461(CAG\u0026gt;GCG/GCA). The 3bp-long mutation is located between the 457th and 464th sites in the NLRP3 3\u0026apos;UTR, and the SNP site of RS 1664775106 (C \u0026gt; G) is at the 457th site (Fig.2).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAssociation between NLRP3 3\u0026rsquo;UTR Polymorphisms and COPD Prevalence\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eIn this study, the gene frequencies of rs10754558 (G\u0026gt;C), rs1664774076 (ATAT\u0026gt;-) and rs1664775106 (C\u0026gt;G) all conform to Hardy-Weinberg genetic balance law and are representative of the population (\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05, Table 4). However, in the study population, there were no homozygous mutation phenotypes in rs1445597018(G/T), rs1664772078(A\u0026gt;C) and rs200307362(A\u0026gt;G), and Hardy-Weinberg statistics were \u003cem\u003eP\u003c/em\u003e\u0026lt; 0.05. This Hardy-Weinberg rule, which depends on allele frequency calculation, when mutation occurred, Therefore, the SNPs of rs1445597018(G\u0026gt;T), rs1664772078(A\u0026gt;C) and rs200307362 (A \u0026gt; G) were not involved in the follow-up study and analysis.\u003c/p\u003e\n\u003cp\u003eThe frequencies of polymorphism of rs1664774076 (ATAT\u0026gt;-) and rs1664775106(C\u0026gt;G) were significantly different between COPD patients and control group.\u0026nbsp;Polymorphisms of rs1664774076 (ATAT\u0026gt;-) and rs1664775106 (C\u0026gt;G) are associated with the occurrence of COPD\u0026nbsp;C\u0026gt;G\u0026nbsp;(rs1664774076\u0026nbsp;ATAT/-\u0026nbsp;vs\u0026nbsp;ATAT/ATAT：OR= 2.221，95%CI = 1.270~3.885，\u003cem\u003eP\u003c/em\u003e= 0.005. \u0026nbsp;-/-\u0026nbsp;vs\u0026nbsp;ATAT/ATAT：OR =3.400，\u0026nbsp;95% CI = 1.952~5.923，\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e 0.0001. Allele - vs Allele ATAT：OR=2.035，95% CI = 1.519~2.728，\u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e0.0001. rs1664775106 CG vs CC：OR=1.852，95%CI = 1.220~2.811，\u003cem\u003eP\u003c/em\u003e=0.004.GG vs CC：OR = 3.409，\u0026nbsp;95% CI = 1.200~9.680，\u003cem\u003e\u0026nbsp;P\u003c/em\u003e = 0.021. Allele G vs Allele C：OR =\u0026nbsp;1.699，95% CI =\u0026nbsp;1.228~2.351，\u003cem\u003eP\u0026nbsp;\u003c/em\u003e=\u0026nbsp;0.001）. However, only C allele at rs10754558 and GCA genotype at 459-461 were associated with the risk of COPD (rs10754558 Allele C vs Allele G: OR = 2.638, 95% CI=1.960~3.550, P \u0026lt; 0.0001). 459-461 GCA vs CAG: OR = 2.381, 95% CI=1.452~3.904, P =0.001) (Table 4).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTABLE 4.\u003c/strong\u003e Genotype frequencies of NLRP3 3UTR gene polymorphisms in control subjects and COPD patients.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\"\u003e\n \u003cp\u003eVariant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\"\u003e\n \u003cp\u003eControls\u003c/p\u003e\n \u003cp\u003e（\u003cem\u003en\u003c/em\u003e=\u0026nbsp;168\u0026nbsp;）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\"\u003e\n \u003cp\u003eTotal COPD\u003c/p\u003e\n \u003cp\u003e（\u003cem\u003en\u003c/em\u003e=221\u0026nbsp;）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\"\u003e\n \u003cp\u003eOR\u0026nbsp;（95% CI）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\"\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalues\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003ers10754558\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e（G\u0026gt;C）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e16（9.52）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e35（15.84）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e73（43.45）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e131（59.28）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e0.820（0.425~1.583）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.555\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e79（47.02）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e55（24.89）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e3.142（1.585~6.229）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eAllele G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e105（31.25）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e201（45.48）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eAllele C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e231（68.75）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e241（54.52）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e2.638（1.960~3.550）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eHWE\u0026nbsp;\u003cem\u003eP\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e0.998\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e0.191\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003ers1664774076\u0026nbsp;（ATAT\u0026gt; -）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eATAT/ATAT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e51（30.36）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e30（13.57）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eATAT/-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e62（36.90）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e81（36.65）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e2.221（1.270~3.885）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e-/-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e55（32.74）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e110（49.77）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e3.400（1.952~5.923）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eAllele ATAT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e164（48.81）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e141（31.90）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eAllele -\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e172（51.19）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e301（68.10）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e2.035（1.519~2.728）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eHWE\u0026nbsp;\u003cem\u003eP\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e0.053\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e0.246\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003ers1664775106\u0026nbsp;（C\u0026gt;G）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e98（58.33）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e92（41.63）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eCG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e65（38.69）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e113（51.13）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.852（1.220~2.811）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e5（2.98）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e16（7.24）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e3.409（1.200~9.680）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.021\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eAllele C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e261（77.68）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e297（67.19）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eAllele G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e75（22.32）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e145（32.81）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.699（1.228~2.351）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eHWE\u0026nbsp;\u003cem\u003eP\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e0.568\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e0.207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e459-461（CAG\u0026gt;GCG/GCA）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eCAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e98（58.33）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e92（41.63）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eGCG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e36（21.43）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e53（23.98）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e1.568（0.942~2.612）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.084\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"25.535714285714285%\" valign=\"top\"\u003e\n \u003cp\u003eGCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.071428571428573%\" valign=\"top\"\u003e\n \u003cp\u003e34（20.24）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.035714285714285%\" valign=\"top\"\u003e\n \u003cp\u003e76（34.39）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e2.381（1.452~3.904）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.357142857142858%\" valign=\"top\"\u003e\n \u003cp\u003e0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn addition, this investigation investigates the difference in allele frequency between COPD subgroups. In severe COPD, the frequency of the C allele in rs10754558 increased. (GOLD1 vs GOLD3-4: \u003cem\u003eP\u0026nbsp;\u003c/em\u003e\u0026lt; 0.05, Fig.3A). The difference of Allele G distribution in rs1664774076 and rs1664775106 is significant in COPD mild to moderate or mild to severe, but there is no difference in allele distribution between moderate and severe (GOLD1 vs GOLD2: \u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e0.001\u003cem\u003e，\u003c/em\u003eGOLD1\u003cem\u003e\u0026nbsp;\u003c/em\u003evs GOLD3-4： \u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e0.001，GOLD2 vs GOLD3-4： \u003cem\u003eP\u0026nbsp;\u003c/em\u003e\u0026gt; 0.05, Fig.3B\u0026amp;3C). The frequency of double mutation GCA and triple mutation GCG genotypes in 459-461 loci increased with the severity of COPD (GOLD1 vs GOLD2: \u003cem\u003eP\u003c/em\u003e＜0.001，GOLD1 vs GOLD3-4： \u003cem\u003eP \u0026lt;\u0026nbsp;\u003c/em\u003e0.001, FIG.2D), and at the same time, the distribution difference between moderate and severe tends to weaken (GOLD2 vs GOLD3-4: P \u0026lt; 0.05, Fig.3D).\u003c/p\u003e\n\u003cp\u003eThe GCG mutations of rs10754558 Allele C, rs1664774076 Allele -, rs1664775106 Allele G, and 459-461 have a low prediction of moderate to severe COPD development risk, and the degree of risk aggravation is insignificant (GOLD2 vs GOLD3-4 \u003cem\u003eP\u003c/em\u003e\u0026gt;0.05). However, the risk of aggravation of mild COPD patients was significantly predicted (GOLD1 vs GOLD2 \u003cem\u003eP\u003c/em\u003e＜0.0001. GOLD1 vs GOLD3-4 \u003cem\u003eP\u003c/em\u003e＜0.0001), except for rs10754558allec (GOLD1 vs GOLD2\u003cem\u003e\u0026nbsp;P\u003c/em\u003e= 0.022. GOLD1 vs GOLD3-4 \u003cem\u003eP\u003c/em\u003e=0.04. In particular, the exposure risk of 459-461 GCA continuous triple mutation was statistically significant among COPD patients of varying severity(GOLD1 vs GOLD2 \u003cem\u003eP\u003c/em\u003e\u003cem\u003e＜\u003c/em\u003e0.0001. GOLD2 vs GOLD3-4 \u003cem\u003eP\u003c/em\u003e= 0.012. GOLD1 vs GOLD3-4 \u003cem\u003eP\u003c/em\u003e＜0.0001) (Table 5).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTABLE 5.\u003c/strong\u003e \u0026nbsp;NLRP3 3\u0026rsquo;UTR\u0026nbsp;SNPs increase the risk of COPD\u0026nbsp;exacerbation\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"701\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eVariant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003eGOLD1\u0026nbsp;VS\u003c/p\u003e\n \u003cp\u003eGOLD 2\u003c/p\u003e\n \u003cp\u003eOR（95% CI）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalues\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003eGOLD2\u0026nbsp;VS\u003c/p\u003e\n \u003cp\u003eGOLD3-4\u003c/p\u003e\n \u003cp\u003eOR（95% CI）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalues\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003eGOLD1\u0026nbsp;VS\u003c/p\u003e\n \u003cp\u003eGOLD3-4\u003c/p\u003e\n \u003cp\u003eOR（95% CI）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalues\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003ers10754558\u0026nbsp;（G/C）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eAllele G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e1.000（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eAllele C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e1.759（1.084~2.853）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e0.022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e0.908（0.571~1.443）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e0.682\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e1.596（1.021~2.497）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e0.040\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003ers1664774076\u0026nbsp;（ATAT\u0026gt; -）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eAllele ATAT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eAllele -\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e3.529（2.100~5.933）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e1.556（0.883~2.741）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e0.126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e5.4903.297~9.142）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003ers1664775106\u0026nbsp;（C\u0026gt;G）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eAllele C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eAllele G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e4.396（2.506~7.710）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e0.710（0.444~1.136）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e0.153\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e3.119（1.825~5.332）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003e459-461（CAG\u0026gt;GCG/GCA）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eCAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e1.000\u0026nbsp;（reference）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eGCG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e31.667（8.776~114.261）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e0.502（0.194~1.296）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e0.154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e15.894（5.065~49.880）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"17.94871794871795%\" valign=\"top\"\u003e\n \u003cp\u003eGCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.65811965811966%\" valign=\"top\"\u003e\n \u003cp\u003e16.077（6.206~41.650）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.25925925925926%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.23931623931624%\" valign=\"top\"\u003e\n \u003cp\u003e0.315（0.127~0.778）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e0.012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"18.66096866096866%\" valign=\"top\"\u003e\n \u003cp\u003e5.059（2.275~11.250）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.116809116809117%\" valign=\"top\"\u003e\n \u003cp\u003e＜0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eDifference of Plasma inflammatory Marker Level by Genotypes of COPD patiants\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe direct downstream factors of NLRP3 are IL-1\u0026beta; and IL-18, while the main activating factor of IL-6 expression is IL-1\u0026beta;. Therefore, we choose to detect the protein levels of NLRP3 and its downstream IL-1\u0026beta;, IL-18 and IL-6 to evaluate the influence of gene changes on transcription and expression. Considering that inducers or exposure factors were required to stimulate the expression of NLRP3 and other proteins, and that these inflammatory factors were all in a low expression state in the control group (Fig.4A), this study investigated whether SNP differences in COPD patients affected the expression of NLRP3 mRNA and downstream factors.\u003c/p\u003e\n\u003cp\u003eThe plasma NLRP3 expression level was higher in the double mutant allele groups rs1664774076 and rs1664775106. At sites 459-461, the plasma NLRP3 expression level increased as the number of mutant bases increased. NLPR3 protein expression was substantially different between the 459-461 GCA and rs1664774076 Allele- groups (\u003cem\u003eP\u003c/em\u003e\u0026lt;0.001). However, the locus of rs10754558 is opposite, and the expression level of plasma NLRP3 in the double mutant allele (CC) group is lower than that in the GG+GC group. The expression levels of inflammatory factors IL-1\u0026beta;, IL-18 and IL-6 downstream of NLRP-3 were basically the same among the groups, but we found that the above inflammatory factors were all elevated in GCA group at 459-461 (NLRP3：GCA vs CAG，\u003cem\u003eP\u003c/em\u003e＜0.001.\u0026nbsp;IL-1\u0026beta;：GCA vs CAG，\u003cem\u003eP\u003c/em\u003e＜0.001.IL-18：GCA vs CAG，\u003cem\u003eP\u003c/em\u003e＜0.05 .\u0026nbsp;IL-6：GCA vs CAG，\u003cem\u003eP\u003c/em\u003e＜0.001), as shown in\u0026nbsp;Fig.4B.\u003c/p\u003e\n\u003cp\u003eIn order to further study whether there are differences in the expression of NLRP3 mRNA in different SNPs, 50 samples including different genotypes in this study were selected for analysis. The transcription level of NLRP3 in each SNPs group is consistent with the expression of NLRP3 protein (Fig.4C).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eNLRP3 3\u0026apos;UTR mutant genotype affects transcription regulation and is related to lung function.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe relative expression of NLRP3 mRNA was negatively correlated with FEV1/FVC%, FEV1%pred, Pa02/PaCO2 (r= -0.7738, -0.6539, -0.4329,\u003cem\u003e\u0026nbsp;P\u003c/em\u003e\u0026lt;0.05). As NLRP3 mRNA levels increased, lung function decreased. Fig.5 demonstrates that FEV1/FVC% has the strongest correlation with the relative expression of mRNA (r = -0.7738), whereas NLRP3 mRNA expression has a moderate correlation with Pa02/PaCO2 (r = -0.4329). From Fig.4C, we know that the relative expression of NLRP3 3\u0026apos;UTR mutant NLRP3 mRNA has increased significantly (rs1664774076 Allele-vs Allele ATAT: \u003cem\u003eP\u003c/em\u003e\u0026lt;0.001. 459-461 GCA vs CAG: \u003cem\u003eP\u003c/em\u003e\u0026lt;0.001), so we analyzed whether the mutant was related to the decline of lung function. rs1664774076 double deletion genotype (-/-) and GCA genotype at 459-461 were negatively correlated with FEV1/FVC% (r= -0.6220, -0.6235, P \u0026lt; 0.001, Fig.6).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eThe predictive value NLRP3 3\u0026rsquo;UTR SNPs for the occurrence and development of COPD\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe diagnostic value of rs1664774076 -/- genotype and GCA genotype at 459-461 locus in predicting the risk of COPD alone or in combination is low (AUC is 0.5852, 0.5708 and 0.5972 respectively). Rs1664774076 -/- genotype is more valuable in predicting the development of COPD than 459-461 GCA genotype (rs1664774076 -/-AUC= 0.7390. 459-461 GCA type AUC= 0.5886), NLRP3 inflammatory factor protein has higher predictive value than its downstream factors IL-6, IL-18 and IL-1\u0026beta; (NLRP3 inflammatory factor protein AUC= 0.9291). The AUC of IL-6, IL-18 and IL-1\u0026beta; were 0.6797, 0.7458 and 0.7841, respectively. The diagnostic value of combining NLRP3 and rs1664774076 -/- was slightly improved (AUC= 0.9301) (Fig.7).\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThere are signaling molecules that activate inflammasomes in the patient\u0026apos;s body, which contribute to the complexity of COPD\u0026apos;s pathogenesis. Pathogen-related molecular pattern stimulation due to infection, such as viruses and bacteria\u003csup\u003e\u0026nbsp;\u003c/sup\u003e[8-9] and damage-related molecular pattern stimulation due to oxidative stress, such as ATP and mitochondrial DNA\u003csup\u003e\u0026nbsp;\u003c/sup\u003e[10] may be the origins of these signaling molecules.This study confirmed the association between NLRP3 and COPD, and found that the expression of NLRP3 and its downstream inflammatory factors was substantially higher in COPD patients than in healthy controls. This distinction is due to two factors: first, the stimulation of external risk factors. Secondly, the presence of SNPs in the NLRP3 population results in variations in inflammatory expression. This can explain why the mutation of the NLRP3 gene does not cause COPD in healthy individuals, and why the development of COPD patients stimulated by the same external factors is heterogeneous.\u003c/p\u003e\n\u003cp\u003eNLRP3 is located in chromosome region 1q43-q44, with 3 kb upstream of the transcription initiation site (exon and intron) and 2 kb downstream of the termination codon (a total of 37,953 kb)\u003csup\u003e\u0026nbsp;\u003c/sup\u003e[11]. Recent genetic research has focused on the association between NLRP3 genetic variation and susceptibility to autoimmune and inflammatory diseases [12-15]. However, they all investigate the polymorphism of the exon and intron genes in the NLRP3 coding region. This study concentrates on the 3\u0026apos; untranslated region (3\u0026apos;UTR) and locates the functional site, which has been confirmed by the population, as opposed to remaining at the level of biological prediction.\u003c/p\u003e\n\u003cp\u003eThere were continuous mutations at sites 459-461 in the 3\u0026apos;UTR region, and the mutant genotypes were GCG or GCA. rs10754558 CC, rs1664774076 Allele- allele (ATAT deletion), and rs1664775106 are examples. COPD susceptibility is associated with the G allele and the 459-461 GCA genotype, which can enhance the risk of COPD. However, for the demonstration that GG is the susceptible genotype\u003csup\u003e\u0026nbsp;\u003c/sup\u003e[11] and other articles\u003csup\u003e\u0026nbsp;\u003c/sup\u003e[16]\u0026nbsp;have reported higher expression of the GG gene NLRP3, one of the possibilities for the difference in results is the regional variation of samples collected, and the effect of the rs10754558 SNP on COPD remains unknown.In addition, the distribution of SNPs differs between subtypes of COPD and is associated with COPD development. Notable is the fact that genotype 459-461 GCA, as a mutant containing three consecutive bases, has the highest degree of mutation, which can not only trigger the early aggravation of risk but also foretell the development risk from GOLD2 to GOLD3-4. Consequently, site mutations are more likely to exacerbate the disease.\u003c/p\u003e\n\u003cp\u003eThis study indicates that SNP mutations can increase the likelihood of disease occurrence or disease development. Polymorphisms in the 3\u0026apos;UTR region influence mRNA stability and translation\u003csup\u003e\u0026nbsp;\u003c/sup\u003e[17], which may substantially affect gene expression by removing, diminishing, or producing miRNA-binding sites. This section of the binding site is also known as the seed region. It consists of 6-7 bases that are complementary to the 3\u0026apos;UTR of the target mRNA. The combination of miRNAs can inhibit the expression of a target protein.\u0026nbsp;The mutants of rs1664774076 (ATAT\u0026gt;-), rs1664775106 (C\u0026gt;G) and 459-461 all showed synchronous up-regulation of NLRP3 mRNA expression and NLRP3 protein level, indicating that there may be a relationship of gene regulation.\u003c/p\u003e\n\u003cp\u003eDifferent SNP genotypes have no effect on the expression of NLRP3\u0026apos;s downstream factors (IL-1\u0026beta;, IL-18, and IL-6). Nevertheless, the 459-461 GCA genotype demonstrated a high level of expression in NLRP3 and its downstream inflammatory factors (\u003cem\u003eP\u003c/em\u003e＜0.05, Fig.4B), which may indicate that the triple base continuous mutation has a greater regulatory influence on the NLRP3 signaling pathway. The mechanism may be that the 459-461 site is located in the seed region of miR-22-3p, and the mutation of three bases results in the loss of the binding site for miR-22-3p, thereby reducing the inhibitory effect of miR-22-3p on the expression of NLRP3 protein and increasing NLRP3. Previous research has also demonstrated that miR-22-3p inhibits the expression of NLRP3 to reduce the inflammatory effects of gout[18-19]. Rs1664774076 (ATAT\u0026gt;-) is recognized by miR-1277-5p and miR-5011-5p, and the mutation at this site corresponds to the deletion of ATAT. Additionally, there is a loss of four bases in the seed region of miRNA, indicating that the mutation at this site may play a larger role in regulating NLRP3. MiR-1277-5p can bind to the 3\u0026apos;UTR of IRS3(Recombinant Insulin Receptor Substrate 3) mRNA to inhibit its expression and reduce cell damage, according to studies [20]. MiR-223-3p has been demonstrated to be a targeted anti-inflammatory therapeutic molecule, and its down-regulation can reduce inflammation [21-23].Rs200307362(A\u0026gt;G) is located in the miR-223-3p-identified seed zone. In contrast, rs200307362 (A\u0026gt;G) did not contain any double mutant GG in this study. It is inferred that the seed region containing rs200307362 can bind tightly to miR-223-3p, and that miR-223-3p inhibits NLRP3.\u003c/p\u003e\n\u003cp\u003eThe level of NLRP3 mRNA increased as lung function declined, and the genotypes of rs1664774076 (ATAT\u0026gt;-) and 459-461 GCA also showed a correlation with lung function decline. The expression levels of NLRP3 mRNA were markedly higher in individuals with the rs1664774076 -/- genotype and the 459-461 GCA genotype. It can be inferred that these two SNP mutations affect the expression of NLRP3, which ultimately reduces COPD patients\u0026apos; lung function and exacerbates the disease.\u003c/p\u003e\n\u003cp\u003eThe rs1664774076 (ATAT\u0026gt;-) and 459-461 GCA genotypes can be used for risk prediction, whereas the detection of the NLRP3 gene has a low diagnostic value for COPD and cannot supplant conventional diagnostic methods. COPD acute exacerbation is measured using inflammation indices (such as CRP, IL-6, and\u0026nbsp;IL-1\u0026beta;)\u003csup\u003e[4]\u0026nbsp;\u003c/sup\u003e.This study discovered that NLRP3 has a greater predictive value than\u0026nbsp;IL-1\u0026beta;\u0026nbsp;and IL-6. In light of the fact that the expression of inflammatory factors has been stimulated by inducing factors, i.e., the expression of inflammatory factors trails behind that of gene detection, it is suggested that the detection of SNPs plus NLRP3 can aid in the clinical evaluation of the progression of a patient\u0026apos;s disease.\u003c/p\u003e\n\u003cp\u003eThis study has some limitations, to conclude. This analysis is limited to the Chinese population and may not be generalizable to other populations. In addition, the scope of this study is limited to the prediction of the miRNA binding site, and it is still necessary to complete the double luciferase reporter gene verification experiment in order to determine the nature of their relationship. In addition, it is necessary to accomplish the miRNA population survey necessary for this study. If the expression level of the miRNA population is minimal, it will also reduce the regulation of SNPs on inflammatory factors.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; Contributions:\u0026nbsp;\u003c/strong\u003e\u003cem\u003eWu Hui Yan\u003c/em\u003e performed miRNAs-bing sites prediction and SNPs screening, data analysis, and article writing. \u003cem\u003eHuang Chu Ting\u003c/em\u003e completed the whole experiment and wrote the original draft. \u003cem\u003eZhang Yan Ling\u003c/em\u003e participated in the screening and enrollment of subjects, blood samples collecting, and clinical data collation. \u003cem\u003ePeng Liang\u003c/em\u003e contributed to project administration. \u003cem\u003eLi Wei Peng\u0026nbsp;\u003c/em\u003eaccomplished the funding acquisition and the writing review of the article. All the authors read and approved the final article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Disclosure Statement:\u0026nbsp;\u003c/strong\u003eNo competing financial interests exist.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eThe research was funded by Medical Scientific Research Foundation of Guangdong Province of China (B2021440).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent Statement:\u0026nbsp;\u003c/strong\u003eWritten informed consent to publish this paper was obtained from the patient(s).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstitutional Review Board Statement:\u0026nbsp;\u003c/strong\u003eThe study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Review Board (or Ethics Committee) of the Fifth Affiliated Hospital of Guangzhou Medical University (protocol KY01-2020-11-06).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration Section:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data that support the SNPs and the sequence of 3\u0026apos;UTR region in NLRP3 of this study are available in NCBI. These miRNA binding sites of NLRP3 were derived from the following resources availiable in the public domain:miRDB, miRTarBase, DIANA, miRcode and TargetScan.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e1. NCBI database\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(The sequence of 3\u0026apos;UTR region in NLRP3 was obtained)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eaccession numbers: Gene ID: 114548 web links:\u003c/p\u003e\n\u003cp\u003ehttps://www.ncbi.nlm.nih.gov/gene/114548#reference-sequences and the matching NCBI blast to verify accession numbers：NG_007509.2 web links:\u003c/p\u003e\n\u003cp\u003ehttps://www.ncbi.nlm.nih.gov/projects/sviewer/?id=NG_007509.2\u0026amp;search=NG_007509.2:g.37562G%3EC\u0026amp;v=1:100\u0026amp;content=5\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2. dbSNP\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e( All SNPs of NLRP3 3\u0026apos;UTR were obtained) accession numbers: search key word: NLRP3 web links:\u003c/p\u003e\n\u003cp\u003ehttps://www.ncbi.nlm.nih.gov/snp/\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3. miRNA prediction websites:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003emiRDB, miRTarBase, DIANA, miRcode and TargetScan\u003c/p\u003e\n\u003cp\u003e3.1 miRDB\u003c/p\u003e\n\u003cp\u003eaccession numbers: search by gene target: NLRP3\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eweb links:\u003c/p\u003e\n\u003cp\u003ehttps://mirdb.org/mirdb/index.html\u003c/p\u003e\n\u003cp\u003e3.2 miRTarBase\u003c/p\u003e\n\u003cp\u003eweb links:\u003c/p\u003e\n\u003cp\u003ehttps://mirtarbase.cuhk.edu.cn/~miRTarBase/miRTarBase_2022/php/search.php?org=hsa\u0026amp;kw=NLRP3\u0026amp;opt=target\u003c/p\u003e\n\u003cp\u003e3.3 DIANA\u003c/p\u003e\n\u003cp\u003eweb links:\u003c/p\u003e\n\u003cp\u003ehttps://dianalab.e-ce.uth.gr/html/dianauniverse/index.php?r=microT_CDS/results\u0026amp;keywords=ENSG00000162711\u0026amp;genes=ENSG00000162711%20\u0026amp;mirnas=\u0026amp;descr=\u0026amp;threshold=0.7\u003c/p\u003e\n\u003cp\u003e3.4 miRcode\u003c/p\u003e\n\u003cp\u003eweb links:\u003c/p\u003e\n\u003cp\u003ehttp://www.mircode.org/index.php?gene=NLRP3\u0026amp;mirfam=\u0026amp;class=\u0026amp;cons=\u0026amp;trregion=\u003c/p\u003e\n\u003cp\u003e3.5 TargetScan\u003c/p\u003e\n\u003cp\u003eweb links:\u003c/p\u003e\n\u003cp\u003ehttps://www.targetscan.org/cgi-bin/targetscan/vert_80/view_gene.cgi?rs=ENST00000391828.3\u0026amp;taxid=9606\u0026amp;showcnc=0\u0026amp;shownc=0\u0026amp;shownc_nc=\u0026amp;showncf1=\u0026amp;showncf2=\u0026amp;subset=1\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eVijayan V K. 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A gain-of-function NLRP3 3\u0026apos;-UTR polymorphism causes miR-146a-mediated suppression of NLRP3 expression and confers protection against sepsis progression[J]. Sci Rep, 2021,11(1):13300.\u003c/li\u003e\n\u003cli\u003eRabe K F, Hurd S, Anzueto A, et al. Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary[J]. Am J Respir Crit Care Med, 2007,176(6):532-555.\u003c/li\u003e\n\u003cli\u003eMartinon F, Agostini L, Meylan E, et al. Identification of bacterial muramyl dipeptide as activator of the NALP3/cryopyrin inflammasome[J]. Curr Biol, 2004,14(21):1929-1934.\u003c/li\u003e\n\u003cli\u003eKanneganti T D, Ozoren N, Body-Malapel M, et al. Bacterial RNA and small antiviral compounds activate caspase-1 through cryopyrin/Nalp3[J]. Nature, 2006,440(7081):233-236.\u003c/li\u003e\n\u003cli\u003eRiteau N, Gasse P, Fauconnier L, et al. Extracellular ATP is a danger signal activating P2X7 receptor in lung inflammation and fibrosis[J]. Am J Respir Crit Care Med, 2010,182(6):774-783.\u003c/li\u003e\n\u003cli\u003eXu G, Huang R, Xia W, et al. Associations between inflammasome-related gene NLRP3 Polymorphisms (rs10754558 and rs35829419) and risk of bladder cancer in a Chinese population[J]. J Clin Lab Anal, 2021,35(11):e23973.\u003c/li\u003e\n\u003cli\u003eWu Z, Wu S, Liang T. Association of NLRP3 rs35829419 and rs10754558 Polymorphisms With Risks of Autoimmune Diseases: A Systematic Review and Meta-Analysis[J]. Front Genet, 2021,12:690860.\u003c/li\u003e\n\u003cli\u003eHitomi Y, Ebisawa M, Tomikawa M, et al. Associations of functional NLRP3 polymorphisms with susceptibility to food-induced anaphylaxis and aspirin-induced asthma[J]. J Allergy Clin Immunol, 2009,124(4):779-785.\u003c/li\u003e\n\u003cli\u003eZhao S, Chen H, Wu G, et al. The association of NLRP3 and TNFRSF1A polymorphisms with risk of ankylosing spondylitis and treatment efficacy of etanercept[J]. J Clin Lab Anal, 2017,31(6).\u003c/li\u003e\n\u003cli\u003eQueiroz G A, Da S R, Pires A O, et al. New variants in NLRP3 inflammasome genes increase risk for asthma and Blomia tropicalis-induced allergy in a Brazilian population[J]. Cytokine X, 2020,2(3):100032.\u003c/li\u003e\n\u003cli\u003eCheng L, Liang X, Qian L, et al. NLRP3 gene polymorphisms and expression in rheumatoid arthritis[J]. Exp Ther Med, 2021,22(4):1110.\u003c/li\u003e\n\u003cli\u003eKim J O, Park H S, Ko E J, et al. The 3\u0026apos;-UTR Polymorphisms in the Thymidylate Synthase (TS) Gene Associated with the Risk of Ischemic Stroke and Silent Brain Infarction[J]. J Pers Med, 2021,11(3).\u003c/li\u003e\n\u003cli\u003eWang X, Chi J, Dong B, et al. MiR-223-3p and miR-22-3p inhibit monosodium urate-induced gouty inflammation by targeting NLRP3[J]. Int J Rheum Dis, 2021,24(4):599-607.\u003c/li\u003e\n\u003cli\u003eZhang Q B, Zhu D, Dai F, et al. MicroRNA-223 Suppresses IL-1beta and TNF-alpha Production in Gouty Inflammation by Targeting the NLRP3 Inflammasome[J]. Front Pharmacol, 2021,12:637415.\u003c/li\u003e\n\u003cli\u003eSun Z, Song L, Li J. Knockdown of small nucleolar RNA host gene 10 (SNHG10) alleviates the injury of human neuroblastoma cells via the miR-1277-5p/insulin substrate receptor 2 axis[J]. Bioengineered, 2022,13(1):709-720.\u003c/li\u003e\n\u003cli\u003eDong H C, Li P N, Chen C J, et al. Sinomenine Attenuates Cartilage Degeneration by Regulating miR-223-3p/NLRP3 Inflammasome Signaling[J]. Inflammation, 2019,42(4):1265-1275.\u003c/li\u003e\n\u003cli\u003eZhao G, Jiang K, Yang Y, et al. The Potential Therapeutic Role of miR-223 in Bovine Endometritis by Targeting the NLRP3 Inflammasome[J]. Front Immunol, 2018,9:1916.\u003c/li\u003e\n\u003cli\u003eNeudecker V, Haneklaus M, Jensen O, et al. 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J Exp Med, 2017,214(6):1737-1752.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"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},"keywords":"Chronic Obstructive Pulmonary Disease, NLRP3, 3'UTR, SNP, miRNA","lastPublishedDoi":"10.21203/rs.3.rs-3605541/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3605541/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjective \u003c/strong\u003eEvaluating the association between a single nucleotide polymorphism in the 3'UTR region of the miRNAs binding site of the NLRP3 gene and the occurrence and development of Chronic Obstructive Pulmonary Disease (COPD) and providing information to aid in the early detection and treatment of COPD.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e The regulatory SNPs located in NLRP3 3'UTR region were searched by using dbSNP database and miRNA binding site prediction database. Meanwhile, samples from COPD patients and healthy controls in the same period were used for verification. The clinical baseline information of all subjects was collected, and the transcription level and protein expression level of NLRP3 and the expression level of inflammatory factors downstream of NLRP3 were detected. The effects of SNPs single nucleotide changes on the transcription and expression of inflammatory factors were analyzed. \u003cstrong\u003eResults\u003c/strong\u003e The study included 418 participants (249 in the COPD group and 169 in the control group). NLRP3 SNPs with miRNA binding sites include rs10754558 (G\u0026gt;C), rs1664774076 (ATAT\u0026gt;-), and rs1664775106 (C\u0026gt;G). Furthermore, two genotypes, GCG and GCA, were discovered to have a linkage mutation at 3'UTR 459-461. COPD susceptibility is tightly associated to the expression of the rs1664774076 -/- genotype, and the risk of COPD increased by 3.4 times (P≤0.0001). Type 459-461 GCA was substantially related with the likelihood of developing COPD at various stages (P\u0026lt;0.05). Except for rs10754558, all homozygous mutants increased NLRP3 mRNA and protein levels. NLRP3 had the greatest area under the ROC curve for predicting the development and diagnosis of COPD when compared to its downstream inflammatory variables (AUC= 0.9291).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e The NLRP3 rs1664774076 -/- genotype is a COPD susceptibility gene, and the GCA genotype at 459-461 can be used as an early predictor of COPD exacerbation. The NLRP3 3'UTR polymorphism may alter the loss of miRNA binding sites, leading to an increase in NLRP3 expression. In the development of COPD, NLRP3 has a better diagnostic value than traditional inflammatory factors.\u003c/p\u003e","manuscriptTitle":"The 3’-UTR Polymorphisms in the NLRP3 Gene Associated with the Risk of COPD and Their Putative Effects on the microRNA Mechanism","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-11-30 17:02:42","doi":"10.21203/rs.3.rs-3605541/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":"8eed210b-dd94-4895-b2ac-720c54e352b0","owner":[],"postedDate":"November 30th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-05-16T10:22:29+00:00","versionOfRecord":[],"versionCreatedAt":"2023-11-30 17:02:42","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3605541","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3605541","identity":"rs-3605541","version":["v1"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}