Exploring potential therapeutic targets for osteoarthritis using proteomics combined with Mendelian randomization

Exploring potential therapeutic targets for osteoarthritis using proteomics combined with Mendelian randomization | 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 Exploring potential therapeutic targets for osteoarthritis using proteomics combined with Mendelian randomization Zihao Zhou, Guanhong Chen This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8016320/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 Background Osteoarthritis (OA) is a common chronic degenerative joint disease. Currently, treatments for the mechanisms underlying OA are lacking. Therefore, his study explored potential therapeutic targets for OA using proteomic and Mendelian randomization (MR) analyses. Methods The knee cartilage of patients with knee osteoarthritis (KOA) and tibial plateau fractures served as the experimental and control groups, respectively. The correlation between the platelet degranulation signaling pathway and KOA pathogenesis was determined using proteomics analysis. We performed a two-sample MR analysis to compare the effects of clopidogrel and aspirin on KOA onset. The literature search results suggested that the ADAMTS family of metalloproteinases is associated with platelet degranulation signaling pathways; therefore, we conducted MR analyses of drug targets to further explore potential therapeutic targets for KOA and extend the screening results to hip osteoarthritis (HOA). Results Proteomics analysis indicated that the platelet degranulation signaling pathway is involved in the pathogenesis of KOA. Following the MR analysis, ADAMTS-1, ADAMTS-5, and ADAMTS-13 were identified as potential targets for OA therapy. Only ADAMTS-13 expression was significantly associated with HOA onset. Conclusion This study provides novel insights into developing targeted treatments for OA, which could significantly improve patient outcomes and quality of life. proteomics mendelian randomization osteoarthritis drug targets pathogenesis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. Introduction Osteoarthritis (OA), a chronic degenerative joint disease prevalent in the general population[1], is characterized by progressive articular cartilage loss, subchondral bone remodeling, and osteophyte formation[2–4]. Globally, the prevalence of OA is high, especially among older adults. Approximately 300 million people in China are affected by OA, with an incidence as high as 62.2% in adults aged over 60 years and 80% in those aged 75 years and above. Although OA has a high incidence rate, its etiology and pathogenesis remain unclear[5]. According to existing studies, the risk factors for OA include heredity, age, sex (female), race, occupation (physical labor), obesity, hypertension, abnormal joint alignment, poor muscle strength, high-intensity exercise, and a history of joint injury. These factors activate cell signaling pathways in joints, including growth factors such as transforming growth factor-β (TGF-β) and Wnt-3a, and signaling molecules such as Smad3, β-catenin, and HIF-2α, thereby triggering pathological changes such as chronic low-grade inflammation, cartilage matrix degradation, and bone hyperplasia[4,6,7,8]. To date, although various drugs and treatment options for alleviating OA symptoms are available, including oral aspirin and other non-steroidal anti-inflammatory drugs, intra-articular injection of sodium hyaluronate, and joint replacement, effective treatments targeting the underlying mechanism of the disease are still lacking[9]. The high incidence of OA not only seriously affects the quality of life of patients but also exerts considerable pressure on medical resources and the social economy[1,3,4]. Therefore, developing targeted drugs for OA treatment is critical. Proteomics technology is among the most effective methods in biological research for studying dynamic changes in protein composition, expression levels, modification state, and interaction in cells. Using this technology, specific biological markers associated with a disease can be identified, and the specific pathogenesis of the disease can be further explored, providing reliable therapeutic targets for precise treatment. Mendelian randomization (MR) is a research method similar to randomized controlled trials, which can determine the causal relationship between risk factors associated with disease using genetic variation index exposure[10]. Therefore, this study utilized a large-scale genome-wide association study (GWAS) dataset, combining MR and drug target MR methods with previous research results of basic experimental and proteomics analysis techniques. This approach aims to further identify key signaling pathways and pathogenic genes in OA pathogenesis, providing new insights for OA treatment. 2. Methods The experimental process is shown in Fig. 1 . 2.1. Proteomics 2.1.1. Experimental sample A total of 26 patients who underwent knee surgery in the orthopedic department of Shan County Central Hospital between January 2023 and December 2023 were selected. Patients with knee OA (KOA) were included in the experimental group, and those with tibial plateau fractures were included in the control group. The experimental group comprised 18 patients (10 females and 8 males) with an average age of 62.5 ± 2.66 years, while 8 patients with an average age of 65.67 ± 3.5 years made up the control group (5 females and 3 males). Written informed consent was obtained from the patients and their families. The study was approved by the Hospital Medical Ethics Committee, with ethical approval number 20230109. Knee cartilage samples were obtained during proteomic analysis. 2.1.2. Inclusion criteria Experimental samples were obtained from patients with KOA who were treated surgically, and control samples were obtained from patients with tibial plateau fractures that could not be reduced during surgery. None of the patients in the experimental or control group had previously undergone knee surgery. The patients in both groups had no contraindications, were suitable for knee surgeries, and had no other serious mental, cardiovascular, or cerebrovascular diseases. Patients who did not meet these criteria were excluded. Additionally, patients with potential confounding conditions, such as rheumatoid and psoriatic arthritis, were excluded from the study after diagnosis. 2.1.3. Protein extraction and mass spectrometry First, an appropriate amount of knee cartilage samples are collected, frozen in liquid nitrogen, and then ground into a fine powder. The powder is subsequently mixed with a lysis buffer and a protease inhibitor cocktail at a 50:1 volume ratio (for example, 1 mL of inhibitor for every 50 mL of lysis buffer). The mixture is thoroughly stirred, followed by sonication for 5 minutes to ensure complete sample lysis. The samples are then centrifuged at 14,000 g for 20 minutes, and 10 µL of the clear supernatant is carefully collected for further analysis. SDS-PAGE electrophoresis is performed to assess protein quality. The gel is stained with Coomassie Brilliant Blue for 30 minutes, followed by multiple decolorization steps until the background is clear and the protein bands are visible. Next, 20 µL of the protein solution is added to 8-well tubes containing MMB magnetic beads and incubated at 37°C for 30 minutes. Subsequently, 45 µL of binding buffer is added, and the mixture is shaken and incubated at room temperature for 15 minutes. After removing the supernatant, the magnetic beads are washed three times with wash buffer. Then, 20 µL of trypsin-containing enzymatic digestion buffer is added, and the reaction is allowed to proceed at 37°C for 4 hours. Following enzymatic digestion, 5 µL of termination buffer is added to stop the reaction, and the mixture is freeze-dried into a powder. For mass spectrometry analysis, two mobile phases are prepared in advance: Phase A is pure water with 0.1% formic acid, and Phase B is 80% acetonitrile mixed with 0.1% formic acid. The freeze-dried sample is dissolved in 10 µL of Phase A and centrifuged at 14,000 g for 20 minutes at 4°C. A 1 µL aliquot of the supernatant is injected into an Orbitrap Eclipse mass spectrometer. The instrument parameters are set as follows: the compensation voltage (CV) switches between − 45 and − 65 V per second, the ion spray voltage is set to 2.0 kV, and the ion transfer tube is maintained at a high temperature of 320°C. After the mass spectrometry analysis, the raw data for proteomic analysis is automatically generated by the instrument. 2.1.4. Data Processing and Quality Control According to the species annotation priority principle, the Homo sapiens UniProt database (version SP, downloaded on March 7, 2023, containing 20,407 protein sequences) was selected for mass spectrometry data matching. The search parameters were set using Proteome Discoverer 2.4 software, including trypsin digestion, with fixed modification of cysteine carbamidomethylation, and dynamic modifications of methionine oxidation and protein N-terminal acetylation. The precursor ion mass tolerance was set to ± 15 ppm, the fragment ion mass tolerance to ± 0.02 Da, and the maximum number of missed cleavages was set to 2. Systematic evaluation of quality control indicators, such as peptide length distribution, number of missed cleavage sites, and peptide-spectrum match (PSM), was performed. Additionally, the physicochemical properties of protein matches, including peptide count, molecular weight, isoelectric point, and sequence coverage, were analyzed to ensure that the data quality met the standards for mass spectrometry detection. Subsequently, bioinformatics techniques are employed to analyze the data and identify key signaling pathways and critical pathogenic genes. 2.1.5. Consumables and Experimental Instruments All reagents and consumables used in this study were sourced from publicly available suppliers. The ammonium bicarbonate, TEAB, and formic acid were purchased from Sigma-Aldrich; urea, dithiothreitol (DTT), and iodoacetamide (IAM) were purchased from Amresco; the protein quantification dye was purchased from Huaxingbio; bovine serum albumin (BSA) was obtained from Thermo Scientific; trypsin was purchased from Promega; acetonitrile was sourced from J.T.Baker; ammonia solution was obtained from Wako Pure Chemical Industries Ltd; and sample vials and caps were purchased from Thermo. The experimental instruments used in this study were also sourced from publicly available suppliers. These include the RIGOL L-3000 high-performance liquid chromatography system purchased from Beijing Puyi Precision Instruments Co., Ltd.; the vortex shaker from SCILOGEX; the vacuum concentrator from Beijing Jiamu Technology Co., Ltd.; the electric constant-temperature water bath purchased from Beijing Guangming Medical Instruments Co., Ltd.; the centrifuge, microplate reader, and electrophoresis system from Shanghai Hefan Instruments Co., Ltd.; and the ultrasonic crusher from Shanghai Huxi Industrial Co., Ltd. 2.2. Two-sample MR analysis To mitigate potential confounding bias caused by racial stratification, our study focused only on participants of European descent. This approach ensured the reliability and consistency of results. The data are all publicly available from GWAS-Catalog, GWAS-IEU, and the FinnGen consortium, and their collection was approved by relevant ethical review committees with informed consent from the participants. Therefore, no further ethical review was required. The experimental process is illustrated in Fig. 2 . The results of basic experimental techniques and proteomics analysis indicated that the key pathways in OA pathogenesis include regulating insulin-like growth factor (IGF)-binding protein (IGFBP) transport, IGF uptake, Post-translational protein phosphorylation, Platelet degranulation, ECM proteoglycans, Regulation of Complement cascade, Degradation of the extracellular matrix, Collagen biosynthesis and modifying enzymes, and Terminal pathway of complements. Since the degranulation of post-translational protein phosphorylation and complement have been confirmed in previous studies[2,3], and the IGFBP-IGF regulation pathway could not be identified as a target in subsequent MR analysis, we have focused on describing the experimental process related to the extracellular matrix and platelet cytoplasmic calcium elevation pathways in this study, and their potential impact on the pathogenesis of KOA. This suggests that extracellular matrix degradation and coagulation may influence the development of KOA[1,11]. Clopidogrel and aspirin are well-known drugs that inhibit platelet granule release activation. Therefore, we will initially use clopidogrel and aspirin as exposure factors and KOA as the outcome factor to investigate the effect of the platelet cytoplasmic calcium elevation pathway on the development of KOA. 2.2.1. Data source Data on the two exposure factors used in the trial (databases using clopidogrel and aspirin) included 462,933(2959 cases and 459974 controls. ID: ukb-b-19698) and 462,933 patients(61702 cases and 401231 controls. ID: ukb-b-8755), respectively, and are publicly available from the IEU OpenGWAS project website ( https://gwas.mrcieu.ac.uk/ ). The KOA database (4,462 cases and 17,885 controls. ID: GCST005813) is publicly available on the official website of GWAS-Catalog ( https://ebi.ac.uk/ ). 2.2.2. Instrumental variable selection The selected instrumental variables (IVs) must meet the three principles of MR. We used clopidogrel and aspirin as significant exposures from GWAS summary data of single nucleotide diversity (single nucleotide polymorphisms, SNPs), with p 10,000 kb was used to ensure the independence of each SNP[12,13,14]. In addition, a correlation analysis was conducted, and SNPs with F > 10 were retained to ensure that the IVs were highly correlated with the exposure factors. Finally, SNPs associated with the results and confounding factors ( P < 5×10 − 8 ) were excluded, and palindromic SNPs with intermediate allele frequencies were removed when reconciling the exposure and outcome data[13]. Since insufficient SNPs were obtained during screening for the IVs of clopidogrel, we relaxed the previous screening criteria to P < 5×10 − 6 . Ultimately, 8 and 13 SNPs were obtained as IVs for using clopidogrel and aspirin, respectively. 2.2.3. Mendelian randomization analysis In this study, we used the ‘TwoSampleMR’ package in R software (version 4.4.1; R Foundation for Statistical Computing, Vienna, Austria) to conduct data analysis[12,13,14]. To verify the causal relationship between the two exposure factors and KOA and further rule out confusion bias, we primarily used inverse variance weighting (IVW) for verification[15]. At the same time, the results from MR-Egger, Weighted Median, Inverse Variance Weighted, Simple Mode, and Weighted Mode were complementary[14]. In addition, Cochran’s Q test was used to assess the heterogeneity of the IVW model, the MR-Egger test was used to determine directed pleiotropy and causal effects[16], and leave-one-out analysis assessed whether a single SNP strongly influenced the causal relationship between exposure and outcome. Outliers were checked and removed using the MR-PRESSO method. At the same time, we plotted funnel plots to enhance the intuitive reliability of our analysis and scatterplots to visually represent the estimated effects[12,13,14,15]. 2.3. Mendelian randomization analysis of drug targets According to the results of two-sample randomization, no obvious causal relationship was observed between the two risk factors and the occurrence of KOA. However, the use of clopidogrel and aspirin appeared to be protective against KOA onset. To further search for potential therapeutic targets, we reviewed relevant literature and found that α-particles are important organelles in the process of platelet degranulation and contain various proteins and molecules involved in the hemostatic response, among which the von Willebrand factor (vWF) is a key hemostatic adhesion glycoprotein[17]. Numerous studies have suggested that a disintegrin and metalloproteinase with thrombospondin protein type 1 motif member 13 (ADAMTS-13) can cleave the vWF[18–24]. Further information on the ADAMTS family has indicated that ADAMTS-1,2, 4, 5, 7, 12, 13, 14, etc., affect KOA pathogenesis[18,25–37]. Moreover, none of the primers related to the ADAMTS family were included in the primers used for our PCR experiments. Therefore, we retrieved the genes encoding these enzymes and their location from the National Library of Medicine ( https://www.ncbi.nlm.nih.gov/gene ) for the target analysis of MR. 2.3.1. Filtering ADAMTS-5 and ADAMTS-13 instrumental variables ADAMTS-1, 2, 4, 5, 7, 12, 13, and 14 were used in MR experiments with drug targets as exposure factors and their corresponding coding genes as drug targets and KOA as the outcome factor. Unfortunately, only ADAMTS-5(ID: GCST90088240) and ADAMTS-13(ID: GCST90088247) met the requirements for the filtering tool variables. Data on drug target exposure factors (5,362 patients for ADAMTS-5 and 5,359 patients for ADAMTS-13) are publicly available from the official website of the GWAS-Catalog ( https://www.ebi.ac.uk/gwas/search ). After the final screening, only the two target genes, ADAMTS-1 and ADAMTS-13, met the experimental requirements. SNPs located within ± 100 kb of ADAMTS-1 and ADAMTS-13, and significantly associated with ADAMTS-1, 2, 4, 5, 7, 12, 13, and 14, were selected as instrumental variables. To further mitigate the impact of strong linkage disequilibrium on the results, we set a threshold of r² < 0.3[38]. Ultimately, 19 SNPs significantly associated with ADAMTS-5 and 15 SNPs significantly associated with ADAMTS-13 were selected, which can be used for target analysis. 2.3.2. Outcome source KOA data were obtained similarly to that of the two-sample MR analysis (containing 22,347 patients) and is publicly available from the official website of the GWAS-Catalog ( https://www.ebi.ac.uk/gwas/search ). 2.3.3. Data analysis ADAMTS-5 and ADAMTS-13 are linked to arthritis and blood clotting, respectively[27]. Therefore, we used the KOA GWAS data as a positive control to verify the IVs. First, we integrated exposure-related drug-targeting IVs into the outcome dataset. Subsequent treatment methods refer to the two-sample MR analysis. 2.3.4. Co-localization analysis The numbers of ADAMTS-5 and ADAMTS-13 in the GWAS database were verified on the official eQTL website ( https://eqtlgen.org/cis-eqtls.html ). Corresponding databases (eqtl-a-ENSG00000154736 and eqtl-a-ENSG00000154734) were obtained from the official IEU OpenGWAS project website ( https://gwas.mrcieu.ac.uk/ ). Using the packages ‘LocusCompareR’ and ‘Gassocplot’ in R software, their co-localization analysis and outcome (KOA) were conducted to determine potential therapeutic targets. 2.4. Extended verification Since OA at different sites, such as HOA and KOA, has similar pathogenesis[11], the selected therapeutic targets may also act on HOA. We selected knee cartilage from patients with KOA and tibial plateau fractures as the experimental specimens for the basic experiment. However, expanding and verifying these results is necessary. Therefore, we obtained the publicly available HOA database (2,396 cases, 9,593 controls. ID: GCST005810.) from the GWAS-Catalog and further validated the effect of ADAMTS family on HOA incidence using drug target MR analysis (the treatment process was the same as above). Co-localization analysis was performed on the selected targets to further explore potential therapeutic targets. 3. Results 3.1. Experimental proteomics results 3.1.1. Screening for differentially expressed proteins Proteomics results were analyzed using a t -test. Differential protein screening was conducted at P < 0.05. Multiples of 1.5 were used as the screening criteria. The results identified 788 differentially expressed proteins, of which 364 were upregulated and 424 were downregulated (Fig. 2 a, 2 b). 3.1.2. Function analysis Response group analysis of the differentially expressed proteins revealed that they were mainly involved in regulating IGFBP transport, IGF uptake, post-translational protein phosphorylation, platelet degranulation, extracellular matrix proteoglycans, complement cascade regulation, extracellular matrix degradation, collagen biosynthesis, modifying enzymes, and the terminal complement pathway (Fig. 2 c). Further analysis of protein–protein interaction showed that the regulation of IGFBP transport, IGF uptake, and platelet degranulation are the key signaling pathways involved in OA pathogenesis. 3.2. Two-sample Mendelian randomization 3.2.1. Instrumental variables After correlation analysis with SNP crossover in GWAS frozen shoulder summary data, exclusion of confounding factors, and removal of outliers using MR-PRESSO, 8 and 13 SNPs were finally identified as IVs using clopidogrel and aspirin, respectively. 3.2.2. Impact of exposure factors on outcome factors The two-sample MR results indicated that while IVW analysis showed that clopidogrel reduced the risk of KOA, no significant causal relationship with KOA incidence was observed(OR[95%] = 0.0000657[1.18×10-13-36688.942], P > 0.05). The effect of aspirin was similar to that of clopidogrel (OR[95%] = 0.4890[0.008–30.364], P > 0.05). The detailed MR-Egger, weighted median, inverse variance weighted, simple mode, and weighted mode analysis results are shown in Table 1 . Table 1 Effect of using clopidogrel and aspirin on the incidence of knee osteoarthritis. Outcome Exposure Method nsnp p val or or_lci95 or_uci95 KOA Clopidogrel MR-Egger 8 0.509223129 8.29E-24 2.73E-88 2.52E + 41 KOA Clopidogrel Weighted median 8 0.880427128 0.143584979 1.50E-12 13790002090 KOA Clopidogrel Inverse variance weighted 8 0.348646727 6.57E-05 1.18E-13 36688.94239 KOA Clopidogrel Simple mode 8 0.960550703 2.71866282 6.70E-17 1.10326E + 17 KOA Clopidogrel Weighted mode 8 0.961409468 2.71866282 2.86E-17 2.58603E + 17 KOA Aspirin MR-Egger 10 0.689371395 9.885143433 0.000195241 500489.8223 KOA Aspirin Weighted median 10 0.785954938 0.578640767 0.011159043 30.00482459 KOA Aspirin Inverse variance weighted 10 0.734177941 0.489056053 0.007876839 30.364443 KOA Aspirin Simple mode 10 0.809978999 0.52103285 0.002991071 90.76188061 KOA Aspirin Weighted mode 10 0.748169356 0.436198449 0.003210299 59.26834587 We performed another Cochran’s Q test on the IVW results of the clopidogrel and aspirin trials and obtained P -values of 0.037 and 0.345 ( P > 0.05), respectively. Funnel (Fig. 3 a, 3 b) and forest (Fig. 3 c, 3 d) plots, leave-one-out analysis (Fig. 3 e, 3 f), and scatter plots (Fig. 3 g, 3 h) of the two trials were plotted. In addition, the P -values for the Egger intercept were 0.57 and 0.58 ( P > 0.05), indicating that the experimental results are relatively reliable. 3.3. Mendelian randomization of drug targets 3.3.1. Instrumental variables Finally, we retained 19 SNPs that were significantly correlated with ADAMTS-5 and could be used to analyze ADAMTS-1 as a target and 15 SNPs that were significantly correlated with ADAMTS-13 and could be used to analyze ADAMTS-13 as a target. 3.3.2. Impact of exposure factors on outcome factors The results of the IVW analysis suggest that the metabolism mediated by ADAMTS-1 and ADAMTS-5 reduces the risk of KOA (OR[95%] = 0.9656[0.926–1.007], P > 0.05). However, the 95% CI for this IVW result includes 1, indicating that there is no significant causal relationship between this metabolic pathway and the development of KOA. On the other hand, the level of ADAMTS-13 appears to increase the risk of KOA (OR[95%] = 1.0491[0.990–1.112], P > 0.05), and its 95% CI also includes 1, suggesting that this result does not reach statistical significance. The detailed MR-Egger, weighted median, IVW, simple mode, and weighted mode analysis results for the two experiments are presented in Table 2 . We also plotted the funnel (Fig. 4 a, 4 b) and forest (Fig. 4 c, 4 d) plots, leave-one-out analyses (Fig. 4 e, 4 f), scatter plots (Fig. 4 g, 4 h), and forest plots of the MR analysis results (Fig. 4 i).The funnel plots for both analyses show that the IVs are evenly distributed on both sides of the line, suggesting that the results are minimally influenced by heterogeneity. Other figures further confirm that the IVW results are not statistically significant, as well as the lack of effect of the two exposures on KOA. Table 2 Drug target Mendelian randomization analysis results of knee osteoarthritis. Outcome Exposure Method nsnp pval or or_lci95 or_uci95 orDrug or_lci95Drug or_uci95Drug KOA ADAMTS-5 MR-Egger 19 0.062980881 0.940574028 0.885483858 0.999091619 1.06318053735051 1.00090920736109 1.12932606343097 KOA ADAMTS-5 Weighted median 19 0.12291055 0.957001832 0.905027957 1.011960458 1.04493007946668 0.988180903866714 1.10493824228111 KOA ADAMTS-5 Inverse variance weighted 19 0.098506516 0.96561772 0.926358372 1.006540891 1.03560651329593 0.993501614025579 1.07949583094822 KOA ADAMTS-5 Simple mode 19 0.561230843 0.974735113 0.895551672 1.060919844 1.02591974673536 0.942578278660135 1.11663015217971 KOA ADAMTS-5 Weighted mode 19 0.144565891 0.957603108 0.905751178 1.012423428 1.04427397096629 0.987729019798239 1.10405597545414 KOA ADAMTS-13 MR Egger 15 0.318449619 1.058437516 0.950748037 1.178324784 0.944788884375749 0.848662451758683 1.05180338094396 KOA ADAMTS-13 Weighted median 15 0.021168671 1.085828277 1.01239978 1.164582481 0.920955938516265 0.858676836253772 0.987752091215969 KOA ADAMTS-13 Inverse variance weighted 15 0.107096416 1.049051376 0.989693424 1.111969387 0.953242160188764 0.89930533328442 1.01041390763548 KOA ADAMTS-13 Simple mode 15 0.130960108 1.098408973 0.979404788 1.231872957 0.91040771192106 0.811772020764687 1.0210282945507 KOA ADAMTS-13 Weighted mode 15 0.049598809 1.090598868 1.007660834 1.180363324 0.916927414167015 0.847196773874284 0.992397408462948 Although the results of both experiments did not reach statistical significance, they indirectly suggest that our experimental findings have good robustness. In both experiments, the P-values for the Cochran Q test in the IVW analysis were 0.980 and 0.240 ( P > 0.05), indicating no significant heterogeneity in the study results. Additionally, the P-values for the Egger intercept were 0.256 and 0.847 ( P > 0.05), suggesting that the study results are relatively reliable. 3.3.3. Co-location analysis Using the ‘LocusCompareR’ and ‘Gassocplot’ packages in R software, ADAMTS-5 and ADAMTS-13 were co-located with the outcome (KOA). We found that rs162489 and rs41314453 were common targets of ADAMTS-1 and ADAMTS-13, respectively, as well as KOA, as shown in Fig. 4 j and Fig. 4 k. 3.4. Expansion verification 3.4.1. Instrumental variables Finally, we selected 19 and 23 SNPs that significantly correlated with ADAMTS-5 and could be used to analyze ADAMTS-1 and ADAMTS-5, respectively. We also selected 15 SNPs that significantly correlated with ADAMTS-13 and could be used to analyze ADAMTS-13. 3.4.2. Impact of exposure factors on outcome factors The results of the IVW analysis suggest that the metabolism mediated by ADAMTS-1 and ADAMTS-5, as well as the level of ADAMTS-5, reduce the risk of HOA, but no significant causal relationship between them and HOA incidence was observed (OR[95%] = 0.9622[0.909–1.018], P > 0.05; OR[95%] = 0.9699[0.908–1.036], P > 0.05). At the same time, there is strong evidence suggesting that ADAMTS-13 increases the risk of developing HOA(OR[95%] = 1.1594[1.079–1.247], P < 0.05). The results of other analyses are shown in Table 3 . Other analyses included funnel (Fig. 5 a, 5 b and 5 c) and forest (Fig. 5 d, 5 e and 5 f) plots, leave-one-out analysis (Fig. 5 g, 5 h and 5 i), scatter plots (Fig. 5 j, 5 k and 5 l), and forest plots of the MR analysis results (Fig. 5 m).The funnel plots for the three analyses also show that the IVs are evenly distributed on both sides of the line, suggesting that the results are minimally influenced by heterogeneity. Furthermore, in the three experiments, the P -values for the Cochran Q test in the IVW analysis were 0.735, 0.988, and 0.818 ( P > 0.05), indicating no significant heterogeneity in the study results. The corresponding P -values for the Egger intercept were 0.73, 0.65, and 0.54 ( P > 0.05), suggesting that the study results are relatively reliable. Table 3 Drug target Mendelian randomization analysis results of hip osteoarthritis. Outcome Exposure Method pval or or_lci95 or_uci95 orDrug or_lci95Drug or_uci95Drug HOA ADAMTS-1 MR-Egger 0.515770248 0.972419879 0.895335025 1.056141438 1.028362358 0.946842879 1.116900347 HOA ADAMTS-1 Weighted median 0.432857992 0.970019967 0.898970496 1.046684781 1.030906614 0.955397478 1.112383559 HOA ADAMTS-1 Inverse variance weighted 0.183911906 0.962227985 0.909102544 1.01845793 1.039254746 0.981876591 1.099985922 HOA ADAMTS-1 Simple mode 0.600571466 0.968671941 0.86167165 1.088959268 1.032341248 0.918307993 1.160534875 HOA ADAMTS-1 Weighted mode 0.251598671 0.958373746 0.893268559 1.028224074 1.04343426 0.972550659 1.119484157 HOA ADAMTS-5 MR-Egger 0.430997928 0.968172256 0.894670354 1.047712727 1.032874051 0.954460106 1.117730117 HOA ADAMTS-5 Weighted median 0.365502565 0.969901604 0.907780756 1.036273479 1.031032422 0.964996229 1.101587574 HOA ADAMTS-5 Inverse variance weighted 0.057435711 0.954105588 0.908967462 1.001485214 1.048102026 0.998516989 1.100149391 HOA ADAMTS-5 Simple mode 0.443974204 0.96010858 0.866697773 1.063587001 1.041548863 0.940214575 1.153804742 HOA ADAMTS-5 Weighted mode 0.414198457 0.970374479 0.904027444 1.041590755 1.030529988 0.960069965 1.106161109 HOA ADAMTS-13 MR-Egger 0.106830707 1.120481629 0.985170887 1.274376962 0.892473356 0.784697174 1.015052326 HOA ADAMTS-13 Weighted median 0.001883136 1.16270397 1.05726347 1.278660013 0.860064148 0.78206872 0.945838032 HOA ADAMTS-13 Inverse variance weighted 6.05E-05 1.159416397 1.078564995 1.246328582 0.862502896 0.802356629 0.927157848 HOA ADAMTS-13 Simple mode 0.005659082 1.258402826 1.096147693 1.444675458 0.794658101 0.692196987 0.912285822 HOA ADAMTS-13 Weighted mode 0.009862616 1.200485389 1.064705121 1.353581514 0.832996394 0.738780775 0.939227191 3.4.3. Co-location analysis Co-localization analysis revealed that rs75152140, rs28744471, and rs78602133 were potential therapeutic targets for ADAMTS-1, ADAMTS-5, and ADAMTS-13, respectively, in the treatment of HOA, as illustrated in Fig. 5 n, Fig. 5 o and Fig. 5 p. 4. Discussion Our experimental results do not explicitly identify ADAMTS-1 and ADAMTS-13 as key targets for the treatment of KOA. Similarly, there is insufficient evidence to support the notion that ADAMTS-1-mediated ADAMTS-5 metabolism and ADAMTS-5 levels play a protective role in the onset of HOA. Additionally, while ADAMTS-13 has been found to be a risk factor for both KOA and HOA, it is considered meaningful only as a therapeutic target for HOA. Although our experimental results did not indicate whether ADAMTS-1 and ADAMTS-5 are associated with the pathogenesis of OA, previous studies have considered both as key proteins in OA development. This discrepancy may be due to the relatively small sample size of OA patients in the GWAS database, which could have introduced bias during the MR analysis. Therefore, future studies may require additional experiments to validate the impact of these two proteins on OA pathogenesis. Interestingly, there are also two important questions for further exploration: Firstly, what is the relationship between ADAMTS-1 and ADAMTS-5? Secondly, why does targeting ADAMTS-13 effectively reduce the risk of HOA? These questions not only stimulate curiosity but also provide crucial directions for future research. In existing studies, ADAMTS-1 and ADAMTS-5 are considered to perform similar functions, namely the cleavage of various proteoglycans, including aggrecan, versican, and brevican[39]. ADAMTS-1 is regarded as a key enzyme responsible for cartilage matrix degradation[2]. Research has shown that ADAMTS-1 can degrade aggrecan, affecting the function and survival of chondrocytes, thus exacerbating cartilage degeneration. In cartilage tissues from OA patients, the expression of ADAMTS-1 is significantly increased, suggesting that it may play a promotive role in OA progression[2,25,28,39]. Additionally, a study by Virtanen et al. identified a genetic linkage between ADAMTS-1 and ADAMTS-5[26], which may explain a potential connection between these two enzymes. ADAMTS-5 is known to degrade key cartilage components, particularly aggrecan[28,29]. Notably, in surgically induced OA models, the activation of ADAMTS-5 is considered one of the primary causes of cartilage destruction[39]. Therefore, precisely targeting and regulating the ADAMTS family may offer a novel strategy for OA treatment. For instance, specific inhibitors targeting ADAMTS-5 have shown potential in delaying the progression of OA in animal models[33]. In previous studies, ADAMTS-13 appears to be a protective factor in the pathogenesis of osteoarthritis (OA). This conclusion may be explained by the following aspects. First, platelet degranulation is a process by which platelets release intracellular particles and various bioactive substances. These substances include platelet growth factor, TGF-β, endogenous tyrosine kinase, etc., which play an important role in injury repair and inflammation[17]. vWF is stored in eccentric nanodomains within platelet α particles and in the Weibel–Palad bodies of endothelial cells[17]. Variations in vWF can promote platelet degranulation and activation, resulting in the failure of fibrinolysis and the possible occurrence of intravascular coagulation and thrombosis. This can, in turn, lead to vascular occlusion and non-vascular necrosis of osteocytes, thus resulting in bone reduction[11]. Second, ADAMTS-13 acts precisely on vWF. When ADAMTS-13 levels increase, large vascular blood factor (ultra large vWF; ULvWF) polymer accumulation in plasma promotes platelet aggregation[18]. Third, platelet activation and thrombosis may lead to thromboinflammation. For example, platelets can form platelet-leukocyte aggregates (PLAs) through interactions with white blood cells, which promote the recruitment and activation of inflammatory cells and intensify the local inflammatory response. In addition, the formation of PLAs is closely associated with microcirculation disorders, which increase the risk of secondary thrombotic events and tissue damage[40]. In addition, activated platelets affect fibrinolysis, leading to intravascular coagulation and thrombosis of the femoral head, eventually leading to vascular occlusion, osteocyte anascular necrosis, osteopenia, and non-traumatic osteonecrosis[11]. Other studies have found that when platelets are subjected to the action of C-reactive protein or thrombin, the activation and release of pro-inflammatory factors such as TNF-α will be triggered, leading to a significant increase in [Ca 2 ⁺] i , which not only promotes platelet adhesion to endothelial cells but also enhances the aggregation ability of platelets, thus aggravating the local inflammatory response[41]. Since OA progression is closely related to the inflammatory state in the synovial membrane, this process may further aggravate joint injury and dysfunction[4,6,7,8]. Therefore, inhibiting platelet activation or blocking the release of inflammatory factors may reduce the symptoms and progression of OA. Fourth, upregulated inflammatory factors such as IL-6 and TNF-α lead to ADAMTS-13 expression downregulation, further aggravating the synovial inflammatory response, thus forming a vicious cycle and promoting further joint damage[22]. Fifth, OA is often accompanied by abnormal synovial angiogenesis, which is closely related to the expression of ADAMTS-13[23]. When ADAMTS-13 activity is reduced, ULvWF accumulation promotes platelet aggregation and microvascular formation, increasing angiogenesis within the synovial tissue. Abnormal angiogenesis not only aggravates the inflammatory response of the synovial membrane but may also lead to insufficient nutrient supply and hypoxia within the joint, further affecting the health of articular cartilage. In addition, the results of an animal study indicated that ADAMTS-13 can reduce vWF-mediated ischemic acute inflammation by reducing IL-6 and TNFα-15, providing a potential therapeutic approach for treating OA by targeting inflammation and pre-thrombotic states[42]. Sixth, ADAMTS-13 may play a role in the progressive loss and degradation of articular cartilage by regulating enzymes associated with the cartilage matrix. Although ADAMTS-13 is not the primary enzyme responsible for directly degrading the cartilage matrix, changes in its activity could indirectly affect the activity of other key proteases, such as ADAMTS-5. However, this appears to contradict the findings of our study. Therefore, we hypothesize that ADAMTS-13 may be involved in the pathogenesis of osteoarthritis through other, as yet undiscovered signaling pathways. Furthermore, in the existing literature, the impact of ADAMTS-13 on the development of OA is rarely mentioned, which may provide a direction for future research. In addition, our study indicates a strong correlation between the IGFBP-IGF-related pathway and OA pathogenesis. However, due to the limited available data, we could not explore the relevant therapeutic targets, which is regrettable. In future studies, deepening our understanding of the influence of IGFBP-IGF-related pathways on OA pathogenesis and exploring novel therapeutic targets may be possible. In the current study, the following arguments may provide new ideas for future research. First, disrupting IGF1/IGF1R signaling significantly affects joint inflammation in rheumatoid arthritis progression. This may lead to a greater perception of pain in patients with rheumatoid arthritis by affecting the homeostasis of chondrocytes, leukocytes, and synovial fibroblasts, as well as the high IGF1R expression in leukocytes[43,44]. Second, other studies have indicated that IGF-1 has an important influence on the biological behavior of chondrocytes. It can regulate cartilage matrix metabolism, promote cartilage repair, and reduce proximal cartilage degeneration at the defect site[45]. Locally directed recombinant adeno-associated virus-mediated IGF-1 gene therapy has shown the potential to enhance osteochondral repair and counteract the effects of early OA[46]. Third, one of the main functions of IGF-1 is to promote growth; simultaneously, it enhances insulin action, inhibits lipolysis, and promotes adipogenesis[47]. IGF-1 therapy may reduce inflammatory responses by reducing the release of high-mobility group protein B1 and the expression of oxidative low-density lipoprotein-induced intercellular adhesion molecules[48]. Fourth, the imaging features of OA are negatively correlated with serum IGF-1 levels, which may vary with age. Serum IGF-1 levels in elderly individuals tend to decline with increasing age[49]. Although the current research on OA has revealed several pathological mechanisms, further research is needed to fully understand its complexity. Future research should focus on the following aspects. 1. Mechanistic studies: Continued exploration of the specific mechanisms of the ADAMTS family, platelet degranulation, and coagulation systems in OA, especially how they interact with synovial inflammation, metabolic disorders, and genetic factors. 2. Therapeutic research: Developing drugs and therapeutic regimens targeting the ADAMTS family, platelet degranulation, and clotting systems, particularly those that can effectively control cartilage matrix degradation and inflammatory responses. 3. Individualized treatment: Based on genetic research results and an in-depth understanding of pathological mechanisms, individualized OA treatment strategies should be developed to improve treatment effects. 4. Preventive measures: Exploring effective preventive measures, such as weight management and early intervention, to reduce the occurrence and progression of OA. 5. Conclusion In our study, proteomics analysis indicated that the key pathways in OA pathogenesis include regulating IGF-binding protein IGFBP transport, IGF uptake, and platelet degranulation signaling pathways. Targeted inhibition of ADAMTS-13 may provide a protective effect against the onset of OA. However, this conclusion is statistically significant only regarding the effect of ADAMTS13 on the onset of HOA. A potential target for achieving this effect is rs78602133. In general, OA treatment must consider various factors, including inflammation, metabolism, genetic factors, and extracellular matrix degradation. An in-depth study of these factors and their interactions could provide new strategies for effectively managing and treating OA, thereby improving the quality of life of patients and reducing the socioeconomic burden. Limitations However, our study has some limitations. First, our proteomics experimental materials were obtained from a Chinese population, whereas the MR analysis was based on data from Europe and America, which may have led to racial bias. Second, a few IVs were used for the two-sample MR analysis and MR target analysis, which may also result in experimental biases. Third, the two-sample MR analysis exhibited heterogeneity, possibly related to different laboratories or measurement methods. Fourth, the outcome factors may contain errors. Because the GWAS data used to analyze OA was obtained from hospital diagnoses, their representation in the population may not be accurate. Declarations Acknowledgements Not applicable. Availability of data and materials The datasets during and/or analysed during the current study available from the corresponding author on reasonable request. Consent for publication Not applicable. Ethics approval All procedures performed in studies involving human participants were in accordance with the ethical standards of the Shan County Central Hospital Research Committee, as well as the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. The study was approved by the Shan County Central Hospital Research Committee (Approval No. 20230109). Clinical trial number Not applicable. Funding Our research has not received any financial support. Competing interests The authors declare that they have no competing interests. Consent to Publish declaration Not applicable. Author contributions Zihao Zhou was responsible for the experimental operations, literature reading, data processing, and article writing. Guanhong Chen guided the experimental procedures and wrote the paper. References W. Jiang, Y. Jin, S. Zhang, Y. Ding, K. Huo, J. Yang, L. Zhao, B. Nian, T. P. Zhong, W. Lu, H. Zhang, X. Cao, K. M. Shah, N. Wang, M. Liu and J. 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16:28:57","extension":"html","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":119120,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/cdd40ba45c929de1c0cb140f.html"},{"id":98421704,"identity":"d04f5cc4-b09a-46d0-a882-38d18583980f","added_by":"auto","created_at":"2025-12-17 16:29:02","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":435989,"visible":true,"origin":"","legend":"\u003cp\u003eExperimental flowchart. OA: osteoarthritis; ADAMTS: A disintegrin and metalloproteinase with thrombospondin motifs; SNP:Single Nucleotide Polymorphism; IV:Instrumental Variable; MR:Mendelian Randomization.\u0026nbsp;\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/9b05e2e270960ee5dc02b6d7.png"},{"id":98422094,"identity":"42217cab-388e-4ef1-8c11-f1f7098c97e1","added_by":"auto","created_at":"2025-12-17 16:30:25","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":196510,"visible":true,"origin":"","legend":"\u003cp\u003eProteomic experiment results.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/a4cb13d96bfff56eb39988b1.png"},{"id":98422920,"identity":"b792b477-136a-41b3-83bf-3eb121206253","added_by":"auto","created_at":"2025-12-17 16:31:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":994815,"visible":true,"origin":"","legend":"\u003cp\u003eTwo-sample Mendelian randomization experiment results.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/fd5f3eb0efeabc8561db25a2.png"},{"id":98422915,"identity":"880b533d-6d0f-4afb-b05d-e36977e5d2d4","added_by":"auto","created_at":"2025-12-17 16:31:38","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":794936,"visible":true,"origin":"","legend":"\u003cp\u003eDrug target Mendelian randomization analysis results of knee osteoarthritis and Co localization analysis.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/1c075e330707136cc7a2b8e3.png"},{"id":98421749,"identity":"a409f24e-2da8-4ec2-bc8b-41ef880e740f","added_by":"auto","created_at":"2025-12-17 16:29:15","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1195691,"visible":true,"origin":"","legend":"\u003cp\u003eDrug target Mendelian randomization analysis results of hip osteoarthritis and Co localization analysis.\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/43cc0f4338dfa41a1859dbdf.png"},{"id":100833447,"identity":"320cf866-b55d-4eee-bcf0-0f8ffdd3b8fe","added_by":"auto","created_at":"2026-01-21 22:24:34","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4972118,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8016320/v1/5ed255f3-ad2f-47f1-b63d-d7f97047759a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Exploring potential therapeutic targets for osteoarthritis using proteomics combined with Mendelian randomization","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eOsteoarthritis (OA), a chronic degenerative joint disease prevalent in the general population[1], is characterized by progressive articular cartilage loss, subchondral bone remodeling, and osteophyte formation[2\u0026ndash;4]. Globally, the prevalence of OA is high, especially among older adults. Approximately 300\u0026nbsp;million people in China are affected by OA, with an incidence as high as 62.2% in adults aged over 60 years and 80% in those aged 75 years and above. Although OA has a high incidence rate, its etiology and pathogenesis remain unclear[5]. According to existing studies, the risk factors for OA include heredity, age, sex (female), race, occupation (physical labor), obesity, hypertension, abnormal joint alignment, poor muscle strength, high-intensity exercise, and a history of joint injury. These factors activate cell signaling pathways in joints, including growth factors such as transforming growth factor-β (TGF-β) and Wnt-3a, and signaling molecules such as Smad3, β-catenin, and HIF-2α, thereby triggering pathological changes such as chronic low-grade inflammation, cartilage matrix degradation, and bone hyperplasia[4,6,7,8]. To date, although various drugs and treatment options for alleviating OA symptoms are available, including oral aspirin and other non-steroidal anti-inflammatory drugs, intra-articular injection of sodium hyaluronate, and joint replacement, effective treatments targeting the underlying mechanism of the disease are still lacking[9].\u003c/p\u003e\u003cp\u003eThe high incidence of OA not only seriously affects the quality of life of patients but also exerts considerable pressure on medical resources and the social economy[1,3,4]. Therefore, developing targeted drugs for OA treatment is critical. Proteomics technology is among the most effective methods in biological research for studying dynamic changes in protein composition, expression levels, modification state, and interaction in cells. Using this technology, specific biological markers associated with a disease can be identified, and the specific pathogenesis of the disease can be further explored, providing reliable therapeutic targets for precise treatment. Mendelian randomization (MR) is a research method similar to randomized controlled trials, which can determine the causal relationship between risk factors associated with disease using genetic variation index exposure[10]. Therefore, this study utilized a large-scale genome-wide association study (GWAS) dataset, combining MR and drug target MR methods with previous research results of basic experimental and proteomics analysis techniques. This approach aims to further identify key signaling pathways and pathogenic genes in OA pathogenesis, providing new insights for OA treatment.\u003c/p\u003e"},{"header":"2. Methods","content":"\u003cp\u003eThe experimental process is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1. Proteomics\u003c/h2\u003e\u003cdiv id=\"Sec4\" class=\"Section3\"\u003e\u003ch2\u003e2.1.1. Experimental sample\u003c/h2\u003e\u003cp\u003eA total of 26 patients who underwent knee surgery in the orthopedic department of Shan County Central Hospital between January 2023 and December 2023 were selected. Patients with knee OA (KOA) were included in the experimental group, and those with tibial plateau fractures were included in the control group. The experimental group comprised 18 patients (10 females and 8 males) with an average age of 62.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.66 years, while 8 patients with an average age of 65.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.5 years made up the control group (5 females and 3 males). Written informed consent was obtained from the patients and their families. The study was approved by the Hospital Medical Ethics Committee, with ethical approval number 20230109. Knee cartilage samples were obtained during proteomic analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section3\"\u003e\u003ch2\u003e2.1.2. Inclusion criteria\u003c/h2\u003e\u003cp\u003eExperimental samples were obtained from patients with KOA who were treated surgically, and control samples were obtained from patients with tibial plateau fractures that could not be reduced during surgery. None of the patients in the experimental or control group had previously undergone knee surgery. The patients in both groups had no contraindications, were suitable for knee surgeries, and had no other serious mental, cardiovascular, or cerebrovascular diseases. Patients who did not meet these criteria were excluded. Additionally, patients with potential confounding conditions, such as rheumatoid and psoriatic arthritis, were excluded from the study after diagnosis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\u003ch2\u003e2.1.3. Protein extraction and mass spectrometry\u003c/h2\u003e\u003cp\u003eFirst, an appropriate amount of knee cartilage samples are collected, frozen in liquid nitrogen, and then ground into a fine powder. The powder is subsequently mixed with a lysis buffer and a protease inhibitor cocktail at a 50:1 volume ratio (for example, 1 mL of inhibitor for every 50 mL of lysis buffer). The mixture is thoroughly stirred, followed by sonication for 5 minutes to ensure complete sample lysis. The samples are then centrifuged at 14,000 g for 20 minutes, and 10 \u0026micro;L of the clear supernatant is carefully collected for further analysis. SDS-PAGE electrophoresis is performed to assess protein quality. The gel is stained with Coomassie Brilliant Blue for 30 minutes, followed by multiple decolorization steps until the background is clear and the protein bands are visible.\u003c/p\u003e\u003cp\u003eNext, 20 \u0026micro;L of the protein solution is added to 8-well tubes containing MMB magnetic beads and incubated at 37\u0026deg;C for 30 minutes. Subsequently, 45 \u0026micro;L of binding buffer is added, and the mixture is shaken and incubated at room temperature for 15 minutes. After removing the supernatant, the magnetic beads are washed three times with wash buffer. Then, 20 \u0026micro;L of trypsin-containing enzymatic digestion buffer is added, and the reaction is allowed to proceed at 37\u0026deg;C for 4 hours. Following enzymatic digestion, 5 \u0026micro;L of termination buffer is added to stop the reaction, and the mixture is freeze-dried into a powder.\u003c/p\u003e\u003cp\u003eFor mass spectrometry analysis, two mobile phases are prepared in advance: Phase A is pure water with 0.1% formic acid, and Phase B is 80% acetonitrile mixed with 0.1% formic acid. The freeze-dried sample is dissolved in 10 \u0026micro;L of Phase A and centrifuged at 14,000 g for 20 minutes at 4\u0026deg;C. A 1 \u0026micro;L aliquot of the supernatant is injected into an Orbitrap Eclipse mass spectrometer. The instrument parameters are set as follows: the compensation voltage (CV) switches between \u0026minus;\u0026thinsp;45 and \u0026minus;\u0026thinsp;65 V per second, the ion spray voltage is set to 2.0 kV, and the ion transfer tube is maintained at a high temperature of 320\u0026deg;C. After the mass spectrometry analysis, the raw data for proteomic analysis is automatically generated by the instrument.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section3\"\u003e\u003ch2\u003e2.1.4. Data Processing and Quality Control\u003c/h2\u003e\u003cp\u003eAccording to the species annotation priority principle, the Homo sapiens UniProt database (version SP, downloaded on March 7, 2023, containing 20,407 protein sequences) was selected for mass spectrometry data matching. The search parameters were set using Proteome Discoverer 2.4 software, including trypsin digestion, with fixed modification of cysteine carbamidomethylation, and dynamic modifications of methionine oxidation and protein N-terminal acetylation. The precursor ion mass tolerance was set to \u0026plusmn;\u0026thinsp;15 ppm, the fragment ion mass tolerance to \u0026plusmn;\u0026thinsp;0.02 Da, and the maximum number of missed cleavages was set to 2. Systematic evaluation of quality control indicators, such as peptide length distribution, number of missed cleavage sites, and peptide-spectrum match (PSM), was performed. Additionally, the physicochemical properties of protein matches, including peptide count, molecular weight, isoelectric point, and sequence coverage, were analyzed to ensure that the data quality met the standards for mass spectrometry detection. Subsequently, bioinformatics techniques are employed to analyze the data and identify key signaling pathways and critical pathogenic genes.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\u003ch2\u003e2.1.5. Consumables and Experimental Instruments\u003c/h2\u003e\u003cp\u003eAll reagents and consumables used in this study were sourced from publicly available suppliers. The ammonium bicarbonate, TEAB, and formic acid were purchased from Sigma-Aldrich; urea, dithiothreitol (DTT), and iodoacetamide (IAM) were purchased from Amresco; the protein quantification dye was purchased from Huaxingbio; bovine serum albumin (BSA) was obtained from Thermo Scientific; trypsin was purchased from Promega; acetonitrile was sourced from J.T.Baker; ammonia solution was obtained from Wako Pure Chemical Industries Ltd; and sample vials and caps were purchased from Thermo.\u003c/p\u003e\u003cp\u003eThe experimental instruments used in this study were also sourced from publicly available suppliers. These include the RIGOL L-3000 high-performance liquid chromatography system purchased from Beijing Puyi Precision Instruments Co., Ltd.; the vortex shaker from SCILOGEX; the vacuum concentrator from Beijing Jiamu Technology Co., Ltd.; the electric constant-temperature water bath purchased from Beijing Guangming Medical Instruments Co., Ltd.; the centrifuge, microplate reader, and electrophoresis system from Shanghai Hefan Instruments Co., Ltd.; and the ultrasonic crusher from Shanghai Huxi Industrial Co., Ltd.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003e2.2. Two-sample MR analysis\u003c/h2\u003e\u003cp\u003eTo mitigate potential confounding bias caused by racial stratification, our study focused only on participants of European descent. This approach ensured the reliability and consistency of results. The data are all publicly available from GWAS-Catalog, GWAS-IEU, and the FinnGen consortium, and their collection was approved by relevant ethical review committees with informed consent from the participants. Therefore, no further ethical review was required. The experimental process is illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe results of basic experimental techniques and proteomics analysis indicated that the key pathways in OA pathogenesis include regulating insulin-like growth factor (IGF)-binding protein (IGFBP) transport, IGF uptake, Post-translational protein phosphorylation, Platelet degranulation, ECM proteoglycans, Regulation of Complement cascade, Degradation of the extracellular matrix, Collagen biosynthesis and modifying enzymes, and Terminal pathway of complements. Since the degranulation of post-translational protein phosphorylation and complement have been confirmed in previous studies[2,3], and the IGFBP-IGF regulation pathway could not be identified as a target in subsequent MR analysis, we have focused on describing the experimental process related to the extracellular matrix and platelet cytoplasmic calcium elevation pathways in this study, and their potential impact on the pathogenesis of KOA. This suggests that extracellular matrix degradation and coagulation may influence the development of KOA[1,11]. Clopidogrel and aspirin are well-known drugs that inhibit platelet granule release activation. Therefore, we will initially use clopidogrel and aspirin as exposure factors and KOA as the outcome factor to investigate the effect of the platelet cytoplasmic calcium elevation pathway on the development of KOA.\u003c/p\u003e\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\u003ch2\u003e2.2.1. Data source\u003c/h2\u003e\u003cp\u003eData on the two exposure factors used in the trial (databases using clopidogrel and aspirin) included 462,933(2959 cases and 459974 controls. ID: ukb-b-19698) and 462,933 patients(61702 cases and 401231 controls. ID: ukb-b-8755), respectively, and are publicly available from the IEU OpenGWAS project website (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://gwas.mrcieu.ac.uk/\u003c/span\u003e\u003cspan address=\"https://gwas.mrcieu.ac.uk/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). The KOA database (4,462 cases and 17,885 controls. ID: GCST005813) is publicly available on the official website of GWAS-Catalog (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://ebi.ac.uk/\u003c/span\u003e\u003cspan address=\"https://ebi.ac.uk/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\u003ch2\u003e2.2.2. Instrumental variable selection\u003c/h2\u003e\u003cp\u003eThe selected instrumental variables (IVs) must meet the three principles of MR. We used clopidogrel and aspirin as significant exposures from GWAS summary data of single nucleotide diversity (single nucleotide polymorphisms, SNPs), with \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;5 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;8\u003c/sup\u003e. The linkage disequilibrium coefficient (\u003cem\u003er\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e) was set to 0.001, and a linkage disequilibrium width\u0026thinsp;\u0026gt;\u0026thinsp;10,000 kb was used to ensure the independence of each SNP[12,13,14]. In addition, a correlation analysis was conducted, and SNPs with F\u0026thinsp;\u0026gt;\u0026thinsp;10 were retained to ensure that the IVs were highly correlated with the exposure factors. Finally, SNPs associated with the results and confounding factors (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;5\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;8\u003c/sup\u003e) were excluded, and palindromic SNPs with intermediate allele frequencies were removed when reconciling the exposure and outcome data[13]. Since insufficient SNPs were obtained during screening for the IVs of clopidogrel, we relaxed the previous screening criteria to \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;5\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;6\u003c/sup\u003e. Ultimately, 8 and 13 SNPs were obtained as IVs for using clopidogrel and aspirin, respectively.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\u003ch2\u003e2.2.3. Mendelian randomization analysis\u003c/h2\u003e\u003cp\u003eIn this study, we used the \u0026lsquo;TwoSampleMR\u0026rsquo; package in R software (version 4.4.1; R Foundation for Statistical Computing, Vienna, Austria) to conduct data analysis[12,13,14]. To verify the causal relationship between the two exposure factors and KOA and further rule out confusion bias, we primarily used inverse variance weighting (IVW) for verification[15]. At the same time, the results from MR-Egger, Weighted Median, Inverse Variance Weighted, Simple Mode, and Weighted Mode were complementary[14]. In addition, Cochran\u0026rsquo;s Q test was used to assess the heterogeneity of the IVW model, the MR-Egger test was used to determine directed pleiotropy and causal effects[16], and leave-one-out analysis assessed whether a single SNP strongly influenced the causal relationship between exposure and outcome. Outliers were checked and removed using the MR-PRESSO method. At the same time, we plotted funnel plots to enhance the intuitive reliability of our analysis and scatterplots to visually represent the estimated effects[12,13,14,15].\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e2.3. Mendelian randomization analysis of drug targets\u003c/h2\u003e\u003cp\u003eAccording to the results of two-sample randomization, no obvious causal relationship was observed between the two risk factors and the occurrence of KOA. However, the use of clopidogrel and aspirin appeared to be protective against KOA onset. To further search for potential therapeutic targets, we reviewed relevant literature and found that α-particles are important organelles in the process of platelet degranulation and contain various proteins and molecules involved in the hemostatic response, among which the von Willebrand factor (vWF) is a key hemostatic adhesion glycoprotein[17]. Numerous studies have suggested that a disintegrin and metalloproteinase with thrombospondin protein type 1 motif member 13 (ADAMTS-13) can cleave the vWF[18\u0026ndash;24]. Further information on the ADAMTS family has indicated that ADAMTS-1,2, 4, 5, 7, 12, 13, 14, etc., affect KOA pathogenesis[18,25\u0026ndash;37]. Moreover, none of the primers related to the ADAMTS family were included in the primers used for our PCR experiments. Therefore, we retrieved the genes encoding these enzymes and their location from the National Library of Medicine (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/gene\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/gene\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) for the target analysis of MR.\u003c/p\u003e\u003cdiv id=\"Sec14\" class=\"Section3\"\u003e\u003ch2\u003e2.3.1. Filtering ADAMTS-5 and ADAMTS-13 instrumental variables\u003c/h2\u003e\u003cp\u003eADAMTS-1, 2, 4, 5, 7, 12, 13, and 14 were used in MR experiments with drug targets as exposure factors and their corresponding coding genes as drug targets and KOA as the outcome factor. Unfortunately, only ADAMTS-5(ID: GCST90088240) and ADAMTS-13(ID: GCST90088247) met the requirements for the filtering tool variables. Data on drug target exposure factors (5,362 patients for ADAMTS-5 and 5,359 patients for ADAMTS-13) are publicly available from the official website of the GWAS-Catalog (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ebi.ac.uk/gwas/search\u003c/span\u003e\u003cspan address=\"https://www.ebi.ac.uk/gwas/search\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAfter the final screening, only the two target genes, ADAMTS-1 and ADAMTS-13, met the experimental requirements. SNPs located within \u0026plusmn;\u0026thinsp;100 kb of ADAMTS-1 and ADAMTS-13, and significantly associated with ADAMTS-1, 2, 4, 5, 7, 12, 13, and 14, were selected as instrumental variables. To further mitigate the impact of strong linkage disequilibrium on the results, we set a threshold of r\u0026sup2; \u0026lt; 0.3[38]. Ultimately, 19 SNPs significantly associated with ADAMTS-5 and 15 SNPs significantly associated with ADAMTS-13 were selected, which can be used for target analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\u003ch2\u003e2.3.2. Outcome source\u003c/h2\u003e\u003cp\u003eKOA data were obtained similarly to that of the two-sample MR analysis (containing 22,347 patients) and is publicly available from the official website of the GWAS-Catalog (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ebi.ac.uk/gwas/search\u003c/span\u003e\u003cspan address=\"https://www.ebi.ac.uk/gwas/search\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section3\"\u003e\u003ch2\u003e2.3.3. Data analysis\u003c/h2\u003e\u003cp\u003eADAMTS-5 and ADAMTS-13 are linked to arthritis and blood clotting, respectively[27]. Therefore, we used the KOA GWAS data as a positive control to verify the IVs. First, we integrated exposure-related drug-targeting IVs into the outcome dataset. Subsequent treatment methods refer to the two-sample MR analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section3\"\u003e\u003ch2\u003e2.3.4. Co-localization analysis\u003c/h2\u003e\u003cp\u003eThe numbers of ADAMTS-5 and ADAMTS-13 in the GWAS database were verified on the official eQTL website (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://eqtlgen.org/cis-eqtls.html\u003c/span\u003e\u003cspan address=\"https://eqtlgen.org/cis-eqtls.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Corresponding databases (eqtl-a-ENSG00000154736 and eqtl-a-ENSG00000154734) were obtained from the official IEU OpenGWAS project website (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://gwas.mrcieu.ac.uk/\u003c/span\u003e\u003cspan address=\"https://gwas.mrcieu.ac.uk/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Using the packages \u0026lsquo;LocusCompareR\u0026rsquo; and \u0026lsquo;Gassocplot\u0026rsquo; in R software, their co-localization analysis and outcome (KOA) were conducted to determine potential therapeutic targets.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003e2.4. Extended verification\u003c/h2\u003e\u003cp\u003eSince OA at different sites, such as HOA and KOA, has similar pathogenesis[11], the selected therapeutic targets may also act on HOA. We selected knee cartilage from patients with KOA and tibial plateau fractures as the experimental specimens for the basic experiment. However, expanding and verifying these results is necessary. Therefore, we obtained the publicly available HOA database (2,396 cases, 9,593 controls. ID: GCST005810.) from the GWAS-Catalog and further validated the effect of ADAMTS family on HOA incidence using drug target MR analysis (the treatment process was the same as above). Co-localization analysis was performed on the selected targets to further explore potential therapeutic targets.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\u003ch2\u003e3.1. Experimental proteomics results\u003c/h2\u003e\u003cdiv id=\"Sec21\" class=\"Section3\"\u003e\u003ch2\u003e3.1.1. Screening for differentially expressed proteins\u003c/h2\u003e\u003cp\u003eProteomics results were analyzed using a \u003cem\u003et\u003c/em\u003e-test. Differential protein screening was conducted at \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05. Multiples of 1.5 were used as the screening criteria. The results identified 788 differentially expressed proteins, of which 364 were upregulated and 424 were downregulated (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea, \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec22\" class=\"Section3\"\u003e\u003ch2\u003e3.1.2. Function analysis\u003c/h2\u003e\u003cp\u003eResponse group analysis of the differentially expressed proteins revealed that they were mainly involved in regulating IGFBP transport, IGF uptake, post-translational protein phosphorylation, platelet degranulation, extracellular matrix proteoglycans, complement cascade regulation, extracellular matrix degradation, collagen biosynthesis, modifying enzymes, and the terminal complement pathway (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec). Further analysis of protein\u0026ndash;protein interaction showed that the regulation of IGFBP transport, IGF uptake, and platelet degranulation are the key signaling pathways involved in OA pathogenesis.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec23\" class=\"Section2\"\u003e\u003ch2\u003e3.2. Two-sample Mendelian randomization\u003c/h2\u003e\u003cdiv id=\"Sec24\" class=\"Section3\"\u003e\u003ch2\u003e3.2.1. Instrumental variables\u003c/h2\u003e\u003cp\u003eAfter correlation analysis with SNP crossover in GWAS frozen shoulder summary data, exclusion of confounding factors, and removal of outliers using MR-PRESSO, 8 and 13 SNPs were finally identified as IVs using clopidogrel and aspirin, respectively.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec25\" class=\"Section3\"\u003e\u003ch2\u003e3.2.2. Impact of exposure factors on outcome factors\u003c/h2\u003e\u003cp\u003eThe two-sample MR results indicated that while IVW analysis showed that clopidogrel reduced the risk of KOA, no significant causal relationship with KOA incidence was observed(OR[95%]\u0026thinsp;=\u0026thinsp;0.0000657[1.18\u0026times;10-13-36688.942], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). The effect of aspirin was similar to that of clopidogrel (OR[95%]\u0026thinsp;=\u0026thinsp;0.4890[0.008\u0026ndash;30.364], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). The detailed MR-Egger, weighted median, inverse variance weighted, simple mode, and weighted mode analysis results are shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of using clopidogrel and aspirin on the incidence of knee osteoarthritis.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"8\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOutcome\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eExposure\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMethod\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ensnp\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cem\u003ep\u003c/em\u003eval\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eor\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eor_lci95\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eor_uci95\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClopidogrel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR-Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.509223129\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e8.29E-24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2.73E-88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.52E\u0026thinsp;+\u0026thinsp;41\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClopidogrel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.880427128\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.143584979\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.50E-12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e13790002090\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClopidogrel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.348646727\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e6.57E-05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.18E-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e36688.94239\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClopidogrel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.960550703\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.71866282\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e6.70E-17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.10326E\u0026thinsp;+\u0026thinsp;17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClopidogrel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.961409468\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.71866282\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2.86E-17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.58603E\u0026thinsp;+\u0026thinsp;17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAspirin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR-Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.689371395\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.885143433\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.000195241\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e500489.8223\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAspirin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.785954938\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.578640767\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.011159043\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e30.00482459\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAspirin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.734177941\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.489056053\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.007876839\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e30.364443\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAspirin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.809978999\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.52103285\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.002991071\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e90.76188061\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAspirin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.748169356\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.436198449\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.003210299\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e59.26834587\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eWe performed another Cochran\u0026rsquo;s Q test on the IVW results of the clopidogrel and aspirin trials and obtained \u003cem\u003eP\u003c/em\u003e-values of 0.037 and 0.345 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), respectively. Funnel (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea, \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb) and forest (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec, \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ed) plots, leave-one-out analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ee, \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ef), and scatter plots (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eg, \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eh) of the two trials were plotted. In addition, the \u003cem\u003eP\u003c/em\u003e-values for the Egger intercept were 0.57 and 0.58 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), indicating that the experimental results are relatively reliable.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e\u003ch2\u003e3.3. Mendelian randomization of drug targets\u003c/h2\u003e\u003cdiv id=\"Sec27\" class=\"Section3\"\u003e\u003ch2\u003e3.3.1. Instrumental variables\u003c/h2\u003e\u003cp\u003eFinally, we retained 19 SNPs that were significantly correlated with ADAMTS-5 and could be used to analyze ADAMTS-1 as a target and 15 SNPs that were significantly correlated with ADAMTS-13 and could be used to analyze ADAMTS-13 as a target.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec28\" class=\"Section3\"\u003e\u003ch2\u003e3.3.2. Impact of exposure factors on outcome factors\u003c/h2\u003e\u003cp\u003eThe results of the IVW analysis suggest that the metabolism mediated by ADAMTS-1 and ADAMTS-5 reduces the risk of KOA (OR[95%]\u0026thinsp;=\u0026thinsp;0.9656[0.926\u0026ndash;1.007], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). However, the 95% CI for this IVW result includes 1, indicating that there is no significant causal relationship between this metabolic pathway and the development of KOA. On the other hand, the level of ADAMTS-13 appears to increase the risk of KOA (OR[95%]\u0026thinsp;=\u0026thinsp;1.0491[0.990\u0026ndash;1.112], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), and its 95% CI also includes 1, suggesting that this result does not reach statistical significance. The detailed MR-Egger, weighted median, IVW, simple mode, and weighted mode analysis results for the two experiments are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. We also plotted the funnel (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb) and forest (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ec, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ed) plots, leave-one-out analyses (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ee, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ef), scatter plots (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eg, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eh), and forest plots of the MR analysis results (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ei).The funnel plots for both analyses show that the IVs are evenly distributed on both sides of the line, suggesting that the results are minimally influenced by heterogeneity. Other figures further confirm that the IVW results are not statistically significant, as well as the lack of effect of the two exposures on KOA.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDrug target Mendelian randomization analysis results of knee osteoarthritis.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOutcome\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eExposure\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMethod\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ensnp\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003epval\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eor\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eor_lci95\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eor_uci95\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eorDrug\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eor_lci95Drug\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u003cp\u003eor_uci95Drug\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR-Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.062980881\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.940574028\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.885483858\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.999091619\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e1.06318053735051\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.00090920736109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.12932606343097\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.12291055\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.957001832\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.905027957\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.011960458\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e1.04493007946668\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.988180903866714\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.10493824228111\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.098506516\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.96561772\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.926358372\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.006540891\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e1.03560651329593\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.993501614025579\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.07949583094822\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.561230843\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.974735113\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.895551672\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.060919844\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e1.02591974673536\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.942578278660135\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.11663015217971\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.144565891\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.957603108\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.905751178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.012423428\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e1.04427397096629\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.987729019798239\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.10405597545414\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.318449619\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.058437516\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.950748037\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.178324784\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.944788884375749\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.848662451758683\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.05180338094396\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.021168671\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.085828277\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.01239978\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.164582481\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.920955938516265\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.858676836253772\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e0.987752091215969\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.107096416\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.049051376\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.989693424\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.111969387\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.953242160188764\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.89930533328442\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.01041390763548\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.130960108\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.098408973\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.979404788\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.231872957\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.91040771192106\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.811772020764687\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e1.0210282945507\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.049598809\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.090598868\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.007660834\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.180363324\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.916927414167015\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.847196773874284\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e\u003cp\u003e0.992397408462948\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eAlthough the results of both experiments did not reach statistical significance, they indirectly suggest that our experimental findings have good robustness. In both experiments, the P-values for the Cochran Q test in the IVW analysis were 0.980 and 0.240 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), indicating no significant heterogeneity in the study results. Additionally, the P-values for the Egger intercept were 0.256 and 0.847 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), suggesting that the study results are relatively reliable.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec29\" class=\"Section3\"\u003e\u003ch2\u003e3.3.3. Co-location analysis\u003c/h2\u003e\u003cp\u003eUsing the \u0026lsquo;LocusCompareR\u0026rsquo; and \u0026lsquo;Gassocplot\u0026rsquo; packages in R software, ADAMTS-5 and ADAMTS-13 were co-located with the outcome (KOA). We found that rs162489 and rs41314453 were common targets of ADAMTS-1 and ADAMTS-13, respectively, as well as KOA, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ej and Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ek.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec30\" class=\"Section2\"\u003e\u003ch2\u003e3.4. Expansion verification\u003c/h2\u003e\u003cdiv id=\"Sec31\" class=\"Section3\"\u003e\u003ch2\u003e3.4.1. Instrumental variables\u003c/h2\u003e\u003cp\u003eFinally, we selected 19 and 23 SNPs that significantly correlated with ADAMTS-5 and could be used to analyze ADAMTS-1 and ADAMTS-5, respectively. We also selected 15 SNPs that significantly correlated with ADAMTS-13 and could be used to analyze ADAMTS-13.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec32\" class=\"Section3\"\u003e\u003ch2\u003e3.4.2. Impact of exposure factors on outcome factors\u003c/h2\u003e\u003cp\u003eThe results of the IVW analysis suggest that the metabolism mediated by ADAMTS-1 and ADAMTS-5, as well as the level of ADAMTS-5, reduce the risk of HOA, but no significant causal relationship between them and HOA incidence was observed (OR[95%]\u0026thinsp;=\u0026thinsp;0.9622[0.909\u0026ndash;1.018], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05; OR[95%]\u0026thinsp;=\u0026thinsp;0.9699[0.908\u0026ndash;1.036], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). At the same time, there is strong evidence suggesting that ADAMTS-13 increases the risk of developing HOA(OR[95%]\u0026thinsp;=\u0026thinsp;1.1594[1.079\u0026ndash;1.247], \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The results of other analyses are shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Other analyses included funnel (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea, \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eb and \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ec) and forest (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ed, \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ee and \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ef) plots, leave-one-out analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eg, \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eh and \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ei), scatter plots (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ej, \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ek and \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003el), and forest plots of the MR analysis results (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003em).The funnel plots for the three analyses also show that the IVs are evenly distributed on both sides of the line, suggesting that the results are minimally influenced by heterogeneity. Furthermore, in the three experiments, the \u003cem\u003eP\u003c/em\u003e-values for the Cochran Q test in the IVW analysis were 0.735, 0.988, and 0.818 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), indicating no significant heterogeneity in the study results. The corresponding \u003cem\u003eP\u003c/em\u003e-values for the Egger intercept were 0.73, 0.65, and 0.54 (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05), suggesting that the study results are relatively reliable.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDrug target Mendelian randomization analysis results of hip osteoarthritis.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"10\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOutcome\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eExposure\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMethod\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003epval\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eor\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eor_lci95\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eor_uci95\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eorDrug\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eor_lci95Drug\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eor_uci95Drug\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR-Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.515770248\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.972419879\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.895335025\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.056141438\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.028362358\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.946842879\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.116900347\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.432857992\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.970019967\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.898970496\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.046684781\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.030906614\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.955397478\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.112383559\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.183911906\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.962227985\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.909102544\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.01845793\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.039254746\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.981876591\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.099985922\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.600571466\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.968671941\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.86167165\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.088959268\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.032341248\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.918307993\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.160534875\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.251598671\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.958373746\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.893268559\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.028224074\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.04343426\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.972550659\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.119484157\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR-Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.430997928\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.968172256\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.894670354\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.047712727\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.032874051\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.954460106\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.117730117\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.365502565\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.969901604\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.907780756\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.036273479\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.031032422\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.964996229\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.101587574\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.057435711\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.954105588\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.908967462\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.001485214\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.048102026\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.998516989\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.100149391\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.443974204\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.96010858\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.866697773\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.063587001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.041548863\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.940214575\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.153804742\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.414198457\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0.970374479\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.904027444\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.041590755\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e1.030529988\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.960069965\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.106161109\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMR-Egger\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.106830707\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.120481629\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.985170887\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.274376962\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.892473356\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.784697174\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e1.015052326\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted median\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.001883136\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.16270397\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.05726347\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.278660013\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.860064148\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.78206872\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.945838032\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eInverse variance weighted\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.05E-05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.159416397\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.078564995\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.246328582\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.862502896\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.802356629\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.927157848\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSimple mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.005659082\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.258402826\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.096147693\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.444675458\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.794658101\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.692196987\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.912285822\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eADAMTS-13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeighted mode\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.009862616\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.200485389\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e1.064705121\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e1.353581514\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.832996394\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e0.738780775\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e\u003cp\u003e0.939227191\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec33\" class=\"Section3\"\u003e\u003ch2\u003e3.4.3. Co-location analysis\u003c/h2\u003e\u003cp\u003eCo-localization analysis revealed that rs75152140, rs28744471, and rs78602133 were potential therapeutic targets for ADAMTS-1, ADAMTS-5, and ADAMTS-13, respectively, in the treatment of HOA, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003en, Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eo \u003cb\u003eand\u003c/b\u003e Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ep.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eOur experimental results do not explicitly identify ADAMTS-1 and ADAMTS-13 as key targets for the treatment of KOA. Similarly, there is insufficient evidence to support the notion that ADAMTS-1-mediated ADAMTS-5 metabolism and ADAMTS-5 levels play a protective role in the onset of HOA. Additionally, while ADAMTS-13 has been found to be a risk factor for both KOA and HOA, it is considered meaningful only as a therapeutic target for HOA. Although our experimental results did not indicate whether ADAMTS-1 and ADAMTS-5 are associated with the pathogenesis of OA, previous studies have considered both as key proteins in OA development. This discrepancy may be due to the relatively small sample size of OA patients in the GWAS database, which could have introduced bias during the MR analysis. Therefore, future studies may require additional experiments to validate the impact of these two proteins on OA pathogenesis. Interestingly, there are also two important questions for further exploration: Firstly, what is the relationship between ADAMTS-1 and ADAMTS-5? Secondly, why does targeting ADAMTS-13 effectively reduce the risk of HOA? These questions not only stimulate curiosity but also provide crucial directions for future research.\u003c/p\u003e\u003cp\u003eIn existing studies, ADAMTS-1 and ADAMTS-5 are considered to perform similar functions, namely the cleavage of various proteoglycans, including aggrecan, versican, and brevican[39]. ADAMTS-1 is regarded as a key enzyme responsible for cartilage matrix degradation[2]. Research has shown that ADAMTS-1 can degrade aggrecan, affecting the function and survival of chondrocytes, thus exacerbating cartilage degeneration. In cartilage tissues from OA patients, the expression of ADAMTS-1 is significantly increased, suggesting that it may play a promotive role in OA progression[2,25,28,39]. Additionally, a study by Virtanen et al. identified a genetic linkage between ADAMTS-1 and ADAMTS-5[26], which may explain a potential connection between these two enzymes. ADAMTS-5 is known to degrade key cartilage components, particularly aggrecan[28,29]. Notably, in surgically induced OA models, the activation of ADAMTS-5 is considered one of the primary causes of cartilage destruction[39]. Therefore, precisely targeting and regulating the ADAMTS family may offer a novel strategy for OA treatment. For instance, specific inhibitors targeting ADAMTS-5 have shown potential in delaying the progression of OA in animal models[33].\u003c/p\u003e\u003cp\u003eIn previous studies, ADAMTS-13 appears to be a protective factor in the pathogenesis of osteoarthritis (OA). This conclusion may be explained by the following aspects. First, platelet degranulation is a process by which platelets release intracellular particles and various bioactive substances. These substances include platelet growth factor, TGF-β, endogenous tyrosine kinase, etc., which play an important role in injury repair and inflammation[17]. vWF is stored in eccentric nanodomains within platelet α particles and in the Weibel\u0026ndash;Palad bodies of endothelial cells[17]. Variations in vWF can promote platelet degranulation and activation, resulting in the failure of fibrinolysis and the possible occurrence of intravascular coagulation and thrombosis. This can, in turn, lead to vascular occlusion and non-vascular necrosis of osteocytes, thus resulting in bone reduction[11]. Second, ADAMTS-13 acts precisely on vWF. When ADAMTS-13 levels increase, large vascular blood factor (ultra large vWF; ULvWF) polymer accumulation in plasma promotes platelet aggregation[18]. Third, platelet activation and thrombosis may lead to thromboinflammation. For example, platelets can form platelet-leukocyte aggregates (PLAs) through interactions with white blood cells, which promote the recruitment and activation of inflammatory cells and intensify the local inflammatory response. In addition, the formation of PLAs is closely associated with microcirculation disorders, which increase the risk of secondary thrombotic events and tissue damage[40]. In addition, activated platelets affect fibrinolysis, leading to intravascular coagulation and thrombosis of the femoral head, eventually leading to vascular occlusion, osteocyte anascular necrosis, osteopenia, and non-traumatic osteonecrosis[11]. Other studies have found that when platelets are subjected to the action of C-reactive protein or thrombin, the activation and release of pro-inflammatory factors such as TNF-α will be triggered, leading to a significant increase in [Ca\u003csup\u003e2\u003c/sup\u003e⁺]\u003csub\u003ei\u003c/sub\u003e, which not only promotes platelet adhesion to endothelial cells but also enhances the aggregation ability of platelets, thus aggravating the local inflammatory response[41]. Since OA progression is closely related to the inflammatory state in the synovial membrane, this process may further aggravate joint injury and dysfunction[4,6,7,8]. Therefore, inhibiting platelet activation or blocking the release of inflammatory factors may reduce the symptoms and progression of OA. Fourth, upregulated inflammatory factors such as IL-6 and TNF-α lead to ADAMTS-13 expression downregulation, further aggravating the synovial inflammatory response, thus forming a vicious cycle and promoting further joint damage[22]. Fifth, OA is often accompanied by abnormal synovial angiogenesis, which is closely related to the expression of ADAMTS-13[23]. When ADAMTS-13 activity is reduced, ULvWF accumulation promotes platelet aggregation and microvascular formation, increasing angiogenesis within the synovial tissue. Abnormal angiogenesis not only aggravates the inflammatory response of the synovial membrane but may also lead to insufficient nutrient supply and hypoxia within the joint, further affecting the health of articular cartilage. In addition, the results of an animal study indicated that ADAMTS-13 can reduce vWF-mediated ischemic acute inflammation by reducing IL-6 and TNFα-15, providing a potential therapeutic approach for treating OA by targeting inflammation and pre-thrombotic states[42]. Sixth, ADAMTS-13 may play a role in the progressive loss and degradation of articular cartilage by regulating enzymes associated with the cartilage matrix. Although ADAMTS-13 is not the primary enzyme responsible for directly degrading the cartilage matrix, changes in its activity could indirectly affect the activity of other key proteases, such as ADAMTS-5. However, this appears to contradict the findings of our study. Therefore, we hypothesize that ADAMTS-13 may be involved in the pathogenesis of osteoarthritis through other, as yet undiscovered signaling pathways. Furthermore, in the existing literature, the impact of ADAMTS-13 on the development of OA is rarely mentioned, which may provide a direction for future research.\u003c/p\u003e\u003cp\u003eIn addition, our study indicates a strong correlation between the IGFBP-IGF-related pathway and OA pathogenesis. However, due to the limited available data, we could not explore the relevant therapeutic targets, which is regrettable. In future studies, deepening our understanding of the influence of IGFBP-IGF-related pathways on OA pathogenesis and exploring novel therapeutic targets may be possible.\u003c/p\u003e\u003cp\u003eIn the current study, the following arguments may provide new ideas for future research. First, disrupting IGF1/IGF1R signaling significantly affects joint inflammation in rheumatoid arthritis progression. This may lead to a greater perception of pain in patients with rheumatoid arthritis by affecting the homeostasis of chondrocytes, leukocytes, and synovial fibroblasts, as well as the high IGF1R expression in leukocytes[43,44]. Second, other studies have indicated that IGF-1 has an important influence on the biological behavior of chondrocytes. It can regulate cartilage matrix metabolism, promote cartilage repair, and reduce proximal cartilage degeneration at the defect site[45]. Locally directed recombinant adeno-associated virus-mediated IGF-1 gene therapy has shown the potential to enhance osteochondral repair and counteract the effects of early OA[46]. Third, one of the main functions of IGF-1 is to promote growth; simultaneously, it enhances insulin action, inhibits lipolysis, and promotes adipogenesis[47]. IGF-1 therapy may reduce inflammatory responses by reducing the release of high-mobility group protein B1 and the expression of oxidative low-density lipoprotein-induced intercellular adhesion molecules[48]. Fourth, the imaging features of OA are negatively correlated with serum IGF-1 levels, which may vary with age. Serum IGF-1 levels in elderly individuals tend to decline with increasing age[49].\u003c/p\u003e\u003cp\u003eAlthough the current research on OA has revealed several pathological mechanisms, further research is needed to fully understand its complexity. Future research should focus on the following aspects. 1. Mechanistic studies: Continued exploration of the specific mechanisms of the ADAMTS family, platelet degranulation, and coagulation systems in OA, especially how they interact with synovial inflammation, metabolic disorders, and genetic factors. 2. Therapeutic research: Developing drugs and therapeutic regimens targeting the ADAMTS family, platelet degranulation, and clotting systems, particularly those that can effectively control cartilage matrix degradation and inflammatory responses. 3. Individualized treatment: Based on genetic research results and an in-depth understanding of pathological mechanisms, individualized OA treatment strategies should be developed to improve treatment effects. 4. Preventive measures: Exploring effective preventive measures, such as weight management and early intervention, to reduce the occurrence and progression of OA.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eIn our study, proteomics analysis indicated that the key pathways in OA pathogenesis include regulating IGF-binding protein IGFBP transport, IGF uptake, and platelet degranulation signaling pathways. Targeted inhibition of ADAMTS-13 may provide a protective effect against the onset of OA. However, this conclusion is statistically significant only regarding the effect of ADAMTS13 on the onset of HOA. A potential target for achieving this effect is rs78602133. In general, OA treatment must consider various factors, including inflammation, metabolism, genetic factors, and extracellular matrix degradation. An in-depth study of these factors and their interactions could provide new strategies for effectively managing and treating OA, thereby improving the quality of life of patients and reducing the socioeconomic burden.\u003c/p\u003e\u003cp\u003e\u003cb\u003eLimitations\u003c/b\u003e\u003c/p\u003e\u003cp\u003eHowever, our study has some limitations. First, our proteomics experimental materials were obtained from a Chinese population, whereas the MR analysis was based on data from Europe and America, which may have led to racial bias. Second, a few IVs were used for the two-sample MR analysis and MR target analysis, which may also result in experimental biases. Third, the two-sample MR analysis exhibited heterogeneity, possibly related to different laboratories or measurement methods. Fourth, the outcome factors may contain errors. Because the GWAS data used to analyze OA was obtained from hospital diagnoses, their representation in the population may not be accurate.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets during and/or analysed during the current study available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll procedures performed in studies involving human participants were in accordance with the ethical standards of the Shan County Central Hospital Research Committee, as well as the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. The study was approved by the Shan County Central Hospital Research Committee (Approval No. 20230109).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOur research has not received any financial support.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003eAuthor contributions\u003c/p\u003e\n\u003cp\u003eZihao Zhou was responsible for the experimental operations, literature reading, data processing, and article writing. Guanhong Chen guided the experimental procedures and wrote the paper.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eW. Jiang, Y. Jin, S. Zhang, Y. Ding, K. Huo, J. Yang, L. Zhao, B. Nian, T. P. Zhong, W. Lu, H. Zhang, X. Cao, K. M. Shah, N. Wang, M. Liu and J. Luo, \u003cem\u003eBone Res.\u003c/em\u003e, \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e10\u003c/em\u003e, 27.\u003c/li\u003e\n\u003cli\u003eC. Y. Yang, A. Chanalaris and L. Troeberg, \u003cem\u003eOsteoarthr. Cartil.\u003c/em\u003e, \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e25,\u003c/em\u003e 1000\u0026ndash;1009.\u003c/li\u003e\n\u003cli\u003eM. Kapoor, J. Martel-Pelletier, D. Lajeunesse, J.-P. Pelletier and H. Fahmi, \u003cem\u003eNat. Rev. Rheumatol.\u003c/em\u003e, \u003cstrong\u003e2011\u003c/strong\u003e, \u003cem\u003e7\u003c/em\u003e, 33\u0026ndash;42.\u003c/li\u003e\n\u003cli\u003eL. Tong, H. Yu, X. Huang, J. Shen, G. Xiao, L. Chen, H. Wang, L. Xing and D. 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Dis.\u003c/em\u003e, \u003cstrong\u003e1993\u003c/strong\u003e, \u003cem\u003e52\u003c/em\u003e, 229\u0026ndash;231.\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":"proteomics, mendelian randomization, osteoarthritis, drug targets, pathogenesis","lastPublishedDoi":"10.21203/rs.3.rs-8016320/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8016320/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eOsteoarthritis (OA) is a common chronic degenerative joint disease. Currently, treatments for the mechanisms underlying OA are lacking. Therefore, his study explored potential therapeutic targets for OA using proteomic and Mendelian randomization (MR) analyses.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eThe knee cartilage of patients with knee osteoarthritis (KOA) and tibial plateau fractures served as the experimental and control groups, respectively. The correlation between the platelet degranulation signaling pathway and KOA pathogenesis was determined using proteomics analysis. We performed a two-sample MR analysis to compare the effects of clopidogrel and aspirin on KOA onset. The literature search results suggested that the ADAMTS family of metalloproteinases is associated with platelet degranulation signaling pathways; therefore, we conducted MR analyses of drug targets to further explore potential therapeutic targets for KOA and extend the screening results to hip osteoarthritis (HOA).\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eProteomics analysis indicated that the platelet degranulation signaling pathway is involved in the pathogenesis of KOA. Following the MR analysis, ADAMTS-1, ADAMTS-5, and ADAMTS-13 were identified as potential targets for OA therapy. Only ADAMTS-13 expression was significantly associated with HOA onset.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThis study provides novel insights into developing targeted treatments for OA, which could significantly improve patient outcomes and quality of life.\u003c/p\u003e","manuscriptTitle":"Exploring potential therapeutic targets for osteoarthritis using proteomics combined with Mendelian randomization","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-11 00:48:27","doi":"10.21203/rs.3.rs-8016320/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":"4676bb44-63c5-4f12-9a59-7ab167288210","owner":[],"postedDate":"December 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-21T22:23:51+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-11 00:48:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8016320","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8016320","identity":"rs-8016320","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}