Genetic and biomechanics insights into cathepsins and non-cancerous digestive diseases: A bidirectional two-sample Mendelian randomization study
Abstract
Background: Digestive diseases have high incidence and mortality rates, posing a significant threat to global health. However, research on these disorders is uneven, while digestive cancers are well-studied and non-cancerous digestive diseases, despite their considerable health impact, have received less attention. Although cathepsins (CTSs), proteases that regulate extracellular matrix (ECM) turnover and cellular stiffness, have been implicated in digestive disorders, their role in disease-specific mechanical perturbations remains unclear. This study bridges this gap by integrating genetic causation with organ-level biomechanics. Methods: To overcome the constraints of conventional epidemiological methods, we employed a dual-sample bidirectional Mendelian randomization (MR) analysis leveraging genome-wide association study (GWAS) data to explore the causal relationships among 9 cathepsins and 23 non-cancerous digestive diseases. We conducted inverse variance weighted (IVW), weighted median (WM), MR-Egger, MR-PRESSO, Cochran’s Q, and sensitivity analyses for thorough evaluation. We also performed correlation analyses to link the biomechanical data with the genetic and disease outcomes, aiming to identify the relationships between mechanical factors, CTSs, and non-cancerous digestive diseases. Results: Forward MR analysis indicated that CTSB promotes both cholecystitis and cholelithiasis and CTSZ promotes chronic gastritis and diverticulosis. Higher CTSL2 levels promote non-alcoholic fatty liver disease (NAFLD) and liver cirrhosis, whereas upregulated CTSG reduces NAFLD risk. Reverse MR analyses indicated that gastroesophageal reflux, gastric ulcer, NAFLD, and cholecystitis elevated CTSE, G, Z, and B levels, respectively; non-alcoholic steatohepatitis elevates CTSB and H levels. Liver cirrhosis increases CTSB, S, and Z; Barrett’s esophagus, celiac disease, and diverticulosis downregulate CTSO, F, and H respectively; chronic pancreatitis lowers CTSE, F, and L2. Multivariable MR analyses revealed the independent effects of individual CTSs on specific diseases: CTSZ as a promoter for diverticulosis, CTSG as a protective factor for NAFLD, and CTSB as a promoter for cholecystitis and cholelithiasis. Conclusion: This study confirmed the causal relationships between cathepsins, mechanical factors in the digestive system, and non-cancerous digestive diseases. By integrating genetic and biomechanical analyses, we have provided a more in-depth understanding of how mechanical forces interact with biological molecules during the development of non-cancerous digestive diseases. Moreover, they may lead to the establishment of novel clinical practice approaches that take into account both the mechanical and biological aspects of digestive diseases, ultimately improving the diagnosis, treatment, and management of these conditions.
References
1. Ricciardiello L. Digestive diseases: Big burden, low funding? Results of the new United European Gastroenterology White Book on digestive diseases. United European Gastroenterology Journal. 2022; 10(7): 627-628. doi: 10.1002/ueg2.12297
2. Rose TC, Pennington A, Kypridemos C, et al. Analysis of the burden and economic impact of digestive diseases and investigation of research gaps and priorities in the field of digestive health in the European Region—White Book 2: Executive summary. United European Gastroenterology Journal. 2022; 10(7): 657-662. doi: 10.1002/ueg2.12298
3. Farthing M, Roberts SE, Samuel DG, et al. Survey of digestive health across Europe: Final report. Part 1: The burden of gastrointestinal diseases and the organisation and delivery of gastroenterology services across Europe. United European Gastroenterology Journal. 2014; 2(6): 539-543. doi: 10.1177/2050640614554154
4. Wang Y, Huang Y, Chase RC, et al. Global Burden of Digestive Diseases: A Systematic Analysis of the Global Burden of Diseases Study, 1990 to 2019. Gastroenterology. 2023; 165(3): 773-783.e15. doi: 10.1053/j.gastro.2023.05.050
5. Hanauer SB. The burdens of digestive diseases. Nature Reviews Gastroenterology & Hepatology. 2009; 6(7): 377-377. doi: 10.1038/nrgastro.2009.104
6. Yadati T, Houben T, Bitorina A, et al. The Ins and Outs of Cathepsins: Physiological Function and Role in Disease Management. Cells. 2020; 9(7): 1679. doi: 10.3390/cells9071679
7. da Costa Fernandes C, Rodríguez VMO, Soares-Costa A, et al. Cystatin-like protein of sweet orange (CsinCPI-2) modulates pre-osteoblast differentiation via β-Catenin involvement. Journal of Materials Science: Materials in Medicine. 2021; 32(4). doi: 10.1007/s10856-021-06504-y
8. Fonović M, Turk B. Cysteine cathepsins and extracellular matrix degradation. Biochimica et Biophysica Acta (BBA) - General Subjects. 2014; 1840(8): 2560-2570. doi: 10.1016/j.bbagen.2014.03.017
9. Vasiljeva O, Reinheckel T, Peters C, et al. Emerging Roles of Cysteine Cathepsins in Disease and their Potential as Drug Targets. Current Pharmaceutical Design. 2007; 13(4): 387-403. doi: 10.2174/138161207780162962
10. de Almeida Chuffa LG, Freire PP, dos Santos Souza J, et al. Aging whole blood transcriptome reveals candidate genes for SARS-CoV-2-related vascular and immune alterations. Journal of Molecular Medicine. 2021; 100(2): 285-301. doi: 10.1007/s00109-021-02161-4
11. Turk B, Turk D, Turk V. Protease signalling: the cutting edge. The EMBO Journal. 2012; 31(7): 1630-1643. doi: 10.1038/emboj.2012.42
12. Patel S, Homaei A, El-Seedi HR, et al. Cathepsins: Proteases that are vital for survival but can also be fatal. Biomedicine & Pharmacotherapy. 2018; 105: 526-532. doi: 10.1016/j.biopha.2018.05.148
13. Barrera C, Ye G, Espejo R, et al. Expression of cathepsins B, L, S, and D by gastric epithelial cells implicates them as antigen presenting cells in local immune responses. Hum Immunol. 2001; 62(10): 1081-1091.
14. Bühling F, Peitz U, Krüger S, et al. Cathepsins K, L, B, X and W are differentially expressed in normal and chronically inflamed gastric mucosa. Biological Chemistry. 2004; 385(5). doi: 10.1515/bc.2004.051
15. Krueger S, Kuester D, Bernhardt A, et al. Regulation of cathepsin X overexpression in H. pylori‐infected gastric epithelial cells and macrophages. The Journal of Pathology. 2008; 217(4): 581-588. doi: 10.1002/path.2485
16. Zdravkova K, Mijanovic O, Brankovic A, et al. Unveiling the Roles of Cysteine Proteinases F and W: From Structure to Pathological Implications and Therapeutic Targets. Cells. 2024; 13(11): 917. doi: 10.3390/cells13110917
17. Iwama H, Mehanna S, Imasaka M, et al. Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis. Scientific Reports. 2021; 11(1). doi: 10.1038/s41598-021-85898-9
18. Aghdassi AA, John DS, Sendler M, et al. Cathepsin D regulates cathepsin B activation and disease severity predominantly in inflammatory cells during experimental pancreatitis. Journal of Biological Chemistry. 2018; 293(3): 1018-1029. doi: 10.1074/jbc.m117.814772
19. Fusco R, Cordaro M, Siracusa R, et al. Biochemical Evaluation of the Antioxidant Effects of Hydroxytyrosol on Pancreatitis-Associated Gut Injury. Antioxidants. 2020; 9(9): 781. doi: 10.3390/antiox9090781
20. Baghy K, Ladányi A, Reszegi A, et al. Insights into the Tumor Microenvironment—Components, Functions and Therapeutics. International Journal of Molecular Sciences. 2023; 24(24): 17536. doi: 10.3390/ijms242417536
21. Mayet WJ, Hermann E, Finsterwalder J, et al. Antibodies to Cathepsin G in Crohn’s disease. European Journal of Clinical Investigation. 1992; 22(6): 427-433. doi: 10.1111/j.1365-2362.1992.tb01485.x
22. Kuwana T, Sato Y, Saka M, et al. Anti-cathepsin G antibodies in the sera of patients with ulcerative colitis. Journal of Gastroenterology. 2000; 35(9): 682-689. doi: 10.1007/s005350070047
23. Yang Z, Liu Y, Qin L, et al. Cathepsin H–Mediated Degradation of HDAC4 for Matrix Metalloproteinase Expression in Hepatic Stellate Cells. The American Journal of Pathology. 2017; 187(4): 781-797. doi: 10.1016/j.ajpath.2016.12.001
24. Fløyel T, Brorsson C, Nielsen LB, et al. CTSH regulates β-cell function and disease progression in newly diagnosed type 1 diabetes patients. Proceedings of the National Academy of Sciences. 2014; 111(28): 10305-10310. doi: 10.1073/pnas.1402571111
25. Steimle A, Gronbach K, Beifuss B, et al. Symbiotic gut commensal bacteria act as host cathepsin S activity regulators. Journal of Autoimmunity. 2016; 75: 82-95. doi: 10.1016/j.jaut.2016.07.009
26. Nørgaard M, Ehrenstein V, Vandenbroucke JP. Confounding in observational studies based on large health care databases: problems and potential solutions – a primer for the clinician. Clinical Epidemiology. 2017; 9: 185-193. doi: 10.2147/clep.s129879
27. Walker V, Sanderson E, Levin MG, et al. Reading and conducting instrumental variable studies: guide, glossary, and checklist. BMJ. 2024; e078093. doi: 10.1136/bmj-2023-078093
28. Burgess S, Davey Smith G, Davies NM, et al. Guidelines for performing Mendelian randomization investigations. Wellcome Open Research. 2019; 4: 186. doi: 10.12688/wellcomeopenres.15555.1
29. Sanderson E, Glymour MM, Holmes MV, et al. Mendelian randomization. Nature Reviews Methods Primers. 2022; 2(1). doi: 10.1038/s43586-021-00092-5
30. Hartwig FP, Davies NM, Hemani G, et al. Two-sample Mendelian randomization: avoiding the downsides of a powerful, widely applicable but potentially fallible technique. International Journal of Epidemiology. 2016; 45(6): 1717-1726. doi: 10.1093/ije/dyx028
31. Sun BB, Maranville JC, Peters JE, et al. Genomic atlas of the human plasma proteome. Nature. 2018; 558(7708): 73-79. doi: 10.1038/s41586-018-0175-2
32. Yavorska OO, Burgess S. MendelianRandomization: an R package for performing Mendelian randomization analyses using summarized data. International Journal of Epidemiology. 2017; 46(6): 1734-1739. doi: 10.1093/ije/dyx034
33. Verbanck M, Chen CY, Neale B, et al. Detection of widespread horizontal pleiotropy in causal relationships inferred from Mendelian randomization between complex traits and diseases. Nature Genetics. 2018; 50(5): 693-698. doi: 10.1038/s41588-018-0099-7
34. Zhu R, Zhang N, Zhu H, et al. Major depressive disorder and the risk of irritable bowel syndrome: A Mendelian randomization study. Molecular Genetics & Genomic Medicine. 2024; 12(3). doi: 10.1002/mgg3.2413
35. Tong T, Zhu C, Farrell JJ, et al. Blood-derived mitochondrial DNA copy number is associated with Alzheimer disease, Alzheimer-related biomarkers and serum metabolites. Alzheimer’s Research & Therapy. 2024; 16(1). doi: 10.1186/s13195-024-01601-w
36. Ma H, Song D, Zhang H, et al. Phenotypic insights into genetic risk factors for immune-related adverse events in cancer immunotherapy. Cancer Immunology, Immunotherapy. 2024; 74(1). doi: 10.1007/s00262-024-03854-8
37. Au Yeung SL, Gill D. Standardizing the reporting of Mendelian randomization studies. BMC Medicine. 2023; 21(1). doi: 10.1186/s12916-023-02894-8
38. Rajamäki K, Lappalainen J, Öörni K, et al. Cholesterol Crystals Activate the NLRP3 Inflammasome in Human Macrophages: A Novel Link between Cholesterol Metabolism and Inflammation. PLoS ONE. 2010; 5(7): e11765. doi: 10.1371/journal.pone.0011765
39. Hornung V, Bauernfeind F, Halle A, et al. Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization. Nature Immunology. 2008; 9(8): 847-856. doi: 10.1038/ni.1631
40. Haasken S, Sutterwala FS. Damage control: Management of cellular stress by the NLRP3 inflammasome. European Journal of Immunology. 2013; 43(8): 2003-2005. doi: 10.1002/eji.201343848
41. Zheng Z, Xiong H, Zhao Z, et al. Tibetan medicine Si-Wei-Qiang-Wei Powder ameliorates cholecystitis via inhibiting the production of pro-inflammatory cytokines and regulating the MAPK signaling pathway. Journal of Ethnopharmacology. 2023; 303: 116026. doi: 10.1016/j.jep.2022.116026
42. Teller A, Jechorek D, Hartig R, et al. Dysregulation of apoptotic signaling pathways by interaction of RPLP0 and cathepsin X/Z in gastric cancer. Pathology - Research and Practice. 2015; 211(1): 62-70. doi: 10.1016/j.prp.2014.09.005
43. Kos J, Sekirnik A, Premzl A, et al. Carboxypeptidases cathepsins X and B display distinct protein profile in human cells and tissues. Experimental Cell Research. 2005; 306(1): 103-113. doi: 10.1016/j.yexcr.2004.12.006
44. Lecaille F, Chazeirat T, Saidi A, et al. Cathepsin V: Molecular characteristics and significance in health and disease. Molecular Aspects of Medicine. 2022; 88: 101086. doi: 10.1016/j.mam.2022.101086
45. Fukuo Y, Yamashina S, Sonoue H, et al. Abnormality of autophagic function and cathepsin expression in the liver from patients with non‐alcoholic fatty liver disease. Hepatology Research. 2014; 44(9): 1026-1036. doi: 10.1111/hepr.12282
46. He Y, Xu M, Zhou C, et al. The Prognostic Significance of CTSV Expression in Patients with Hepatocellular Carcinoma. International Journal of General Medicine. 2024; 17: 4867-4881. doi: 10.2147/ijgm.s467179
47. Manchanda M, Das P, Gahlot GPS, et al. Cathepsin L and B as Potential Markers for Liver Fibrosis: Insights From Patients and Experimental Models. Clinical and Translational Gastroenterology. 2017; 8(6): e99. doi: 10.1038/ctg.2017.25
48. Toonen EJM, Mirea AM, Tack CJ, et al. Activation of Proteinase 3 Contributes to Nonalcoholic Fatty Liver Disease and Insulin Resistance. Molecular Medicine. 2016; 22(1): 202-214. doi: 10.2119/molmed.2016.00033
49. Zamolodchikova TS, Tolpygo SM, Svirshchevskaya EV. Cathepsin G—Not Only Inflammation: The Immune Protease Can Regulate Normal Physiological Processes. Frontiers in Immunology. 2020; 11. doi: 10.3389/fimmu.2020.00411
50. Davies NM, Holmes MV, Davey Smith G. Reading Mendelian randomisation studies: a guide, glossary, and checklist for clinicians. BMJ. 2018; k601. doi: 10.1136/bmj.k601
51. Ai S, Zhang J, Zhao G, et al. Causal associations of short and long sleep durations with 12 cardiovascular diseases: linear and nonlinear Mendelian randomization analyses in UK Biobank. European Heart Journal. 2021; 42(34): 3349-3357. doi: 10.1093/eurheartj/ehab170
52. Yang Q, Sanderson E, Tilling K, et al. Exploring and mitigating potential bias when genetic instrumental variables are associated with multiple non-exposure traits in Mendelian randomization. European Journal of Epidemiology. 2022; 37(7): 683-700. doi: 10.1007/s10654-022-00874-5
53. Li J, Tang M, Gao X, et al. Mendelian randomization analyses explore the relationship between cathepsins and lung cancer. Communications Biology. 2023; 6(1). doi: 10.1038/s42003-023-05408-7
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