The ClinicianThe ClinicianКлиницист1818-83382412-8775Publishing House ABV Press66510.17650/1818-8338-2025-19-4-K751ORIGINAL INVESTIGATIONSОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕResearch ArticleFunctional analysis of gene-gene interaction networks of miR-143-3p and miR-181b-5p as regulators of coronary atherosclerotic plaque vulnerabilityФункциональный анализ сетей ген-генных взаимодействий miR-143-3p и miR-181b-5p как регуляторов стабильности коронарных атеросклеротических бляшекhttps://orcid.org/0000-0002-8209-2791ShchekochikhinD. Yu.ЩекочихинДмитрий Юрьевич
Russian Federation
rozhkov_a_n@staff.sechenov.ruhttps://orcid.org/0000-0002-2735-076XRozhkovAndrey N.РожковАндрей Николаевич
Russian Federation
rozhkov_a_n@staff.sechenov.ruhttps://orcid.org/0009-0007-6667-1287ErshovaN. A.ЕршоваНаталья Алексеевна
Russian Federation
rozhkov_a_n@staff.sechenov.ruhttps://orcid.org/0000-0001-5124-6383KopylovF. Yu.КопыловФилипп Юрьевич
Russian Federation
rozhkov_a_n@staff.sechenov.ruI.M. Sechenov First Moscow State Medical University, Ministry of Health of Russia (Sechenov University)ФГАОУ ВО Первый Московский государственный медицинский университет им. И. М. Сеченова Минздрава России (Сеченовский Университет)Institute of Personalized Cardiology of the “Digital Biodesign and Personalized Healthcare” Center, Science and Technology Park of I. M. Sechenov First Moscow State Medical University, Ministry of Health of Russia (Sechenov University)Институт персонализированной кардиологии Центра «Цифровой биодизайн и персонализированное здравоохранение» Научно-технологического парка биомедицины ФГАОУ ВО Первый Московский государственный медицинский университет им. И. М. Сеченова Минздрава России (Сеченовский Университет)17032026194536205102025Copyright ©; 2026, Shchekochikhin D.Y., Rozhkov A.N., Ershova N.A., Kopylov F.Y.Copyright ©; 2026, Щекочихин Д.Ю., Рожков А.Н., Ершова Н.А., Копылов Ф.Ю.2026Shchekochikhin D.Y., Rozhkov A.N., Ershova N.A., Kopylov F.Y.Щекочихин Д.Ю., Рожков А.Н., Ершова Н.А., Копылов Ф.Ю.https://creativecommons.org/licenses/by/4.0https://klinitsist.abvpress.ru/Klin/article/view/665

Aim. To perform functional analysis of the gene-gene interaction networks of miR-143-3p and miR-181b-5p to identify key target genes involved in regulating the coronary atherosclerotic plaque vulnerability and to identify potential therapeutic targets.

Material and methods. The study is based on the analysis of a database of 62 patients with suspected coronary artery atherosclerosis examined by multi-slice computed tomography coronary angiography. The sample included 22 patients with vulnerable plaques, 23 with stable plaques, and 17 without atherosclerosis. Analysis of circulating microRNA levels in blood plasma was performed via real-time reverse transcription polymerase chain reaction using TaqMan kits. Data on experimentally validated target genes were obtained from the TarBase v9.0 database, and data on gene interactions were obtained from the STRING v12.0 database. Interaction networks were built using CytoScape software.

Results. Analysis of the functional connections of miR-143-3p revealed an effect on the key genes HNF4A, SMAD3 and AKT1 involved in the regulation of lipid metabolism, proliferation and differentiation of vascular smooth muscle cells, and angiogenesis. For miR-181b-5p, the main target genes VCAM1, ARRB2, BCL2, and IGF1R were identified, which regulate the processes of monocyte adhesion, inflammation, apoptosis, and vascular stability. Network analysis demonstrated the convergence of PI3K / AKT, TGF-β / SMAD signaling pathways and cell death control mechanisms in regulation of atherosclerotic plaque stability.

Conclusion. Functional analysis revealed the key molecular targets of miR-143-3p and miR-181b-5p involved in pathogenesis of atherosclerosis. The identified microRNAs and their target genes represent promising therapeutic targets for stabilizing atherosclerotic plaques and preventing cardiovascular complications. Evaluation of the expression levels of these microRNAs in combination with imaging techniques can contribute to the development of personalized approaches to diagnosis and treatment of atherosclerosis.

Цель исследования – провести функциональный анализ сетей ген-генных взаимодействий микроРНК miR-143-3p и miR-181b-5p для выявления ключевых генов-мишеней, участвующих в регуляции стабильности коронарных атеросклеротических бляшек, и определения потенциальных терапевтических мишеней.

Материал и методы. В основе исследования лежит анализ базы данных 62 пациентов с подозрением на атеросклероз коронарных артерий, обследованных методом мультиспиральной компьютерной томографии коронарных артерий. Выборка включала 22 пациента с уязвимыми бляшками, 23 – со стабильными бляшками и 17 – без атеросклероза. Анализ уровней циркулирующих микроРНК в плазме крови проводили методом полимеразной цепной реакции в реальном времени с обратной транскрипцией с использованием наборов TaqMan. Данные об экспериментально валидированных генах-мишенях получали из базы данных TarBase v.9.0, а данные о взаимодействиях генов – из базы STRING v.12.0. Построение сетей взаимодействия осуществляли с помощью программного обеспечения CytoScape.

Результаты. Анализ функциональных связей микроРНК miR-143-3p выявил ее ключевое влияние на гены HNF4A, SMAD3 и AKT1, участвующие в регуляции метаболизма липидов, пролиферации и дифференцировки гладкомышечных клеток сосудов и ангиогенеза. Для miR-181b-5p идентифицированы основные гены-мишени VCAM1, ARRB2, BCL2 и IGF1R, регулирующие процессы адгезии моноцитов, воспаления, апоптоза и стабильности сосудистой стенки. Сетевой анализ продемонстрировал конвергенцию сигнальных путей PI3K / AKT, TGF-β / SMAD и механизмов контроля клеточной гибели в регуляции стабильности атеросклеротических бляшек.

Заключение. Функциональный анализ выявил ключевые молекулярные мишени микроРНК miR-143-3p и miR-181b-5p, участвующие в патогенезе атеросклероза. Идентифицированные микроРНК и их гены-мишени представляют собой перспективные терапевтические мишени для стабилизации атеросклеротических бляшек и предотвращения сердечно-сосудистых осложнений. Профилирование экспрессии данных микроРНК в сочетании с визуализационными методами может способствовать развитию персонализированных подходов к диагностике и лечению атеросклероза.

microRNAsmiR-143-3pmiR-181b-5patherosclerosisplaque stabilityVCAM1HNF4ASMAD3AKT1ARRB2IGF1RBCL2микроРНКmiR-143-3pmiR-181b-5pатеросклерозстабильность бляшекVCAM1HNF4ASMAD3AKT1ARRB2IGF1RBCL2The study was performed with the support from the Ministry of Science and Higher Education of the Russian Federation in the framework of state support of development of worlds-class research centers “Digital biodesign and personalized healthcare” No. 075-15-2022-304.Исследование проведено при поддержке Министерства науки и высшего образования Российской Федерации в рамках государственной поддержки создания и развития исследовательских центров мирового уровня «Цифровой биодизайн и персонализированное здравоохранение» № 075-15-2022-304.Teixeira A.R., Ferreira V.V., Pereira-da-Silva T. et al. The role of miRNAs in the diagnosis of stable atherosclerosis of different arterial territories: a critical review. Front Cardiovasc Med 2022;9:1040971. DOI: 10.3389/fcvm.2022.1040971Huang P. Potential new therapeutic targets: association of microRNA with atherosclerotic plaque stability. Int J Immunopathol Pharmacol 2023;37:3946320231185657. DOI: 10.1177/03946320231185657Churov A., Summerhill V., Grechko A. et al. MicroRNAs as potential biomarkers in atherosclerosis. Int J Mol Sci 2019;20(22):5547. DOI: 10.3390/ijms20225547Rozhkov A.N., Shchekochikhin D.Y., Ashikhmin Y.I. et al. The profile of circulating blood microRNAs in outpatients with vulnerable and stable atherosclerotic plaques: associations with cardiovascular risks. Noncoding RNA 2022;8(4):47. DOI: 10.3390/ncrna8040047González-López P., Ares-Carral C., López-Pastor A.R. et al. Implication of miR-155-5p and miR-143-3p in the vascular insulin resistance and instability of human and experimental atherosclerotic plaque. Int J Mol Sci 2022;23(18):10253. DOI: 10.3390/ijms231810253Zhou H., Yang D., Cheng H.S. et al. miR-181b regulates vascular endothelial aging by modulating an MAP3K3 signaling pathway. FASEB J 2022;36(6):e22353. DOI: 10.1096/fj.202200046RChang L., Zhou G., Soufan O., Xia J. MiRNet 2.0: network-based visual analytics for miRNA functional analysis and systems biology. Nucleic Acids Res 2020;48(W1):W244–51. DOI: 10.1093/nar/gkaa467Lu H., Lei X., Winkler R. et al. Crosstalk of hepatocyte nuclear factor 4a and glucocorticoid receptor in the regulation of lipid metabolism in mice fed a high-fat-high-sugar diet. Lipids Health Dis 2022;21(1):46. DOI: 10.1186/s12944-022-01654-6Burdin D.V., Kolobov A.A., Brocker C. et al. Diabetes-linked transcription factor HNF4α regulates metabolism of endogenous methylarginines and β-aminoisobutyric acid by controlling expression of alanine-glyoxylate aminotransferase 2. Sci Rep 2016;6:35503. DOI: 10.1038/srep35503Cheng P., Wirka R.C., Kim J.B. et al. Smad3 regulates smooth muscle cell fate and mediates adverse remodeling and calcification of the atherosclerotic plaque. Nat Cardiovasc Res 2022;1(4):322–33. DOI: 10.1038/s44161-022-00042-8Petrovič D., Letonja J., Petrovič D. SMAD3 rs17228212 polymorphism is associated with advanced carotid atherosclerosis in a slovenian population. Biomedicines 2024;12(5):1103. DOI: 10.3390/biomedicines12051103Yang S., Li J., Chen Y. et al. MicroRNA-216a promotes M1 macrophages polarization and atherosclerosis progression by activating telomerase via the Smad3/NF-κB pathway. Biochim Biophys Acta Mol Basis Dis 2019;1865(7):1772–81. DOI: 10.1016/j.bbadis.2018.06.016Wang Y., Li Y., Liu D. Erythropoietin promoted intraplaque angiogenesis by PI3K/AKT/mTOR signaling pathway in atherosclerosis. Tissue Cell 2023;82:102084. DOI: 10.1016/j.tice.2023.102084Zhao Y., Qian Y., Sun Z. et al. Role of PI3K in the progression and regression of atherosclerosis. Front Pharmacol 2021;12:632378. DOI: 10.3389/fphar.2021.632378Pickett J.R., Wu Y., Zacchi L.F. et al. Targeting endothelial vascular cell adhesion molecule-1 in atherosclerosis: drug discovery and development of vascular cell adhesion molecule-1-directed novel therapeutics. Cardiovasc Res 2023;119(13):2278–93. DOI: 10.1093/cvr/cvad130Wang Y., Fan Z., Xu C. et al. Anti-ATR001 monoclonal antibody ameliorates atherosclerosis through beta-arrestin2 pathway. Biochem Biophys Res Commun 2021;544:1–7. DOI: 10.1016/j.bbrc.2021.01.054Drozd M., Bruns A., Yuldasheva N. et al. Endothelial insulin-like growth factor-1 signaling regulates vascular barrier function and atherogenesis. Cardiovasc Res 2025;121(7):1108–20. DOI: 10.1093/cvr/cvaf055Kutuk O., Basaga H. Bcl-2 protein family: implications in vascular apoptosis and atherosclerosis. Apoptosis 2006;11(10):1661–75. DOI: 10.1007/s10495-006-9402-7Lv B., He S., Li P. et al. MicroRNA-181 in cardiovascular disease: emerging biomarkers and therapeutic targets. FASEB J 2024;38(9):e23635. DOI: 10.1096/fj.202400306RWu J., Zhu Y., Liu D. et al. Biological functions and potential mechanisms of miR-143-3p in cancers (review). Oncol Rep 2024;52(3):113. DOI: 10.3892/or.2024.8772Stoffel M., Duncan S.A. The maturity-onset diabetes of the young (MODY1) transcription factor HNF4alpha regulates expression of genes required for glucose transport and metabolism. Proc Natl Acad Sci USA 1997;94(24):13209–14. DOI: 10.1073/pnas.94.24.13209Ferguson J.F., Matthews G.J., Townsend R.R. et al. Candidate gene association study of coronary artery calcification in chronic kidney disease: findings from the CRIC study (Chronic Renal Insufficiency Cohort). J Am Coll Cardiol 2013;62(9):789–98. DOI: 10.1016/j.jacc.2013.01.103McDonald T.J., Ellard S. Maturity onset diabetes of the young: identification and diagnosis. Ann Clin Biochem 2013;50(Pt 5): 403–15. DOI: 10.1177/0004563213483458Thymiakou E., Othman A., Hornemann T. et al. Defects in high density lipoprotein metabolism and hepatic steatosis in mice with liver-specific ablation of hepatocyte nuclear factor 4A. Metabolism 2020;110:154307. DOI: 10.1016/j.metabol.2020.154307Temtem M., Serrao M., Mendonca M.I. et al. Is HNF4A gene, a risk factor or protection against coronary artery disease? Eur Heart J 2021;42. DOI: 10.1093/eurheartj/ehab724.3196Yang X., Chen L., Xu X. et al. TGF-beta/Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage. J Cell Biol 2001;153(1):35–46. DOI: 10.1083/jcb.153.1.35Miller C.L., Pjanic M., Wang T. et al. Integrative functional genomics identifies regulatory mechanisms at coronary artery disease loci. Nat Commun 2016;7:12092. DOI: 10.1038/ncomms12092Nikpay M., Goel A., Won H.-H. et al. A comprehensive 1000 genomes-based genome-wide association meta-analysis of coronary artery disease. Nat Genet 2015;47(10):1121–30. DOI: 10.1038/ng.3396Fang Y., Duan C., Chen S. et al. Increased legumain/Smad3 expression in atherosclerotic plaque of rat thoracic aorta. Biomed Pharmacother 2019;119:109353. DOI: 10.1016/j.biopha.2019.109353Chen J., Somanath P.R., Razorenova O. et al. Akt1 regulates pathological angiogenesis, vascular maturation and permeability in vivo. Nat Med 2005;11(11):1188–96. DOI: 10.1038/nm1307Ackah E., Yu J., Zoellner S. et al. Akt1/protein kinase B alpha is critical for ischemic and VEGF-mediated angiogenesis. J Clin Invest 2005;115(8):2119–27. DOI: 10.1172/JCI24726Linton M.F., Moslehi J.J., Babaev V.R. Akt signaling in macrophage polarization, survival, and atherosclerosis. Int J Mol Sci 2019;20(10):2703. DOI: 10.3390/ijms20112703Liu Y.-S., Lin H.-Y., Lai S.-W. et al. MiR-181b modulates EGFR-dependent VCAM-1 expression and monocyte adhesion in glioblastoma. Oncogene 2017;36(35):5006–22. DOI: 10.1038/onc.2017.129Ghasempour G., Mohammadi A., Zamani-Garmsiri F. et al. MiRNAs through β-ARR2/p-ERK1/2 pathway regulate the VSMC proliferation and migration. Life Sci 2021;279:119703. DOI: 10.1016/j.lfs.2021.119703Ley K., Huo Y. VCAM-1 is critical in atherosclerosis. J Clin Invest 2001;107(10):1209–10. DOI: 10.1172/JCI13005Troncoso M.F., Ortiz-Quintero J., Garrido-Moreno V. et al. VCAM-1 as a predictor biomarker in cardiovascular disease. Biochim Biophys Acta Mol Basis Dis 2021;1867(9):166170. DOI: 10.1016/j.bbadis.2021.166170Cybulsky M.I., Iiyama K., Li H. et al. A major role for VCAM-1, but not ICAM-1, in early atherosclerosis. J Clin Invest 2001;107(10):1255–62. DOI: 10.1172/JCI11871Weinkauf C.C., Concha-Moore K., Lindner J.R. et al. Endothelial vascular cell adhesion molecule 1 is a marker for high-risk carotid plaques and target for ultrasound molecular imaging. J Vasc Surg 2018;68(6):105S–13. DOI: 10.1016/j.jvs.2017.10.088Wang H., Deng Q.W., Peng A.N. et al. β-Arrestin2 functions as a key regulator in the sympathetic-triggered immunodepression after stroke. J Neuroinflammation 2018;15(1):102. DOI: 10.1186/s12974-018-1142-4Kuhr F.K., Zhang Y., Brovkovych V. et al. Beta-arrestin 2 is required for B1 receptor-dependent post-translational activation of inducible nitric oxide synthase. FASEB J 2010;24(7):2475–83. DOI: 10.1096/fj.09-148783Kim J., Zhang L., Peppel K. et al. Beta-arrestins regulate atherosclerosis and neointimal hyperplasia by controlling smooth muscle cell proliferation and migration. Circ Res 2008;103(1):70–9. DOI: 10.1161/circresaha.108.172338Cao Z.H., Gao L., Jiang L. et al. Effect of β-arrestin on damage of human umbilical vein endothelial cell induced by angiotensin II. Eur Rev Med Pharmacol Sci 2017;21(24):5821–6. DOI: 10.26355/eurrev_201712_14030Verma S., Kumar A., Narang R. et al. Signature transcriptome analysis of stage specific atherosclerotic plaques of patients. BMC Med Genomics 2022;15(1):99. DOI: 10.1186/s12920-022-01250-8Ruvolo P.P., Deng X., May W.S. Phosphorylation of Bcl2 and regulation of apoptosis. Leukemia 2001;15(4):515–22. DOI: 10.1038/sj.leu.2402090Klimentova E.A., Suchkov I.A., Shchulkin A.V. et al. Expression of apoptotic markers Bcl-2 and Bax in the vascular wall. Sovremennye tekhnologii v meditsine = Sovrem Tekhnologii Med 2021;13(2):46–50. (In Russ.). DOI: 10.17691/stm2021.13.2.05Климентова Э.А., Сучков И.А., Щулькин А.П. и др. Экспрессия маркеров апоптоза Всl-2 и Вах в сосудистой стенке. Современные технологии в медицине 2021;13(2):46–50. DOI: 10.17691/stm2021.13.2.05Qian Y., Teng Y., Li Y. et al. MiR-143-3p suppresses the progression of nasal squamous cell carcinoma by targeting Bcl-2 and IGF1R. Biochem Biophys Res Commun 2019;518(3):492–9. DOI: 10.1016/j.bbrc.2019.08.075Chang L., Chai X., Chen P. et al. MiR-181b-5p suppresses starvation-induced cardiomyocyte autophagy by targeting Hspa5. Int J Mol Med 2019;43(1):143–54. DOI: 10.3892/ijmm.2018.3988Fagerberg L., Hallström B.M., Oksvold P. et al. Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol Cell Proteomics 2014;13(2):397–406. DOI: 10.1074/mcp.M113.035600Higashi Y., Sukhanov S., Shai S.Y. et al. Insulin-like growth factor-1 receptor deficiency in macrophages accelerates atherosclerosis and induces an unstable plaque phenotype in apolipoprotein E-deficient mice. Circulation 2016;133(23):2263–78. DOI: 10.1161/circulationaha.116.021805Okura Y., Brink M., Zahid A.A. et al. Decreased expression of insulin-like growth factor-1 and apoptosis of vascular smooth muscle cells in human atherosclerotic plaque. J Mol Cell Cardiol 2001;33(10):1777–89. DOI: 10.1006/jmcc.2001.1441Yang Z., Wang J., Pan Z. et al. MiR-143-3p regulates cell proliferation and apoptosis by targeting IGF1R and IGFBP5 and regulating the Ras/p38 MAPK signaling pathway in rheumatoid arthritis. Exp Ther Med 2018;15(4):3781–90. DOI: 10.3892/etm.2018.5907Mahmoudian M., Razmara E., Mahmud Hussen B. et al. Identification of a six-microRNA signature as a potential diagnostic biomarker in breast cancer tissues. J Clin Lab Anal 2021;35(11):e24010. DOI: 10.1002/jcla.24010Than M., Han M. Functional analysis of the miRNA–mRNA interaction network in C. elegans. Worm 2013;2(4):e26894. DOI: 10.4161/worm.26894