miR-19a-3p enhances TGF-β1-induced cardiac fibroblast activation via targeting BAMBI

Abstract

Myocardial fibrosis is a major pathogenic factor contributing to cardiac remodeling and heart failure. Recent research has indicated that microRNAs play a crucial role in the progression of cardiac fibrosis. Bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) have been shown to alleviate myocardial fibrosis by inhibiting the transforming growth factor β1 (TGF-β1) signaling pathway. Therefore, the current study aimed to elucidate the post-transcriptional regulation of BAMBI by miR-19a-3p and its role in TGF-β1-induced cardiac fibroblast activation. We found that transverse aortic constriction induced both myocardial interstitial and perivascular collagen deposition. Quantitative reverse transcription-PCR (qRT-PCR) analysis showed that the expression level of miR-19a-3p was increased in the myocardial tissues of cardiac fibrosis, and TGF-β1 induced an upregulation in miR-19a-3p expression in cardiac fibroblasts. The dual-luciferase reporter assay and qRT-PCR verified that miR-19a-3p directly bound to the 3′ untranslated regions of BAMBI mRNA, thereby reducing BAMBI expression and diminishing its ability to inhibit the TGF-β1 signaling pathway. Furthermore, overexpression of miR-19a-3p mimic increased the activation of TGF-β1/SMAD2/3 pathway signaling, promoting cardiac fibroblast activation. However, this activation was blocked by BAMBI overexpression. These findings imply that miR-19a-3p enhances the activation of TGF-β1/SMAD2/3 by inhibiting BAMBI, further boosting the activation of cardiac fibroblasts and contributing to myocardial fibrosis.

Keywords

miR-19a-3p / BAMBI / TGF-β1 / SMAD2/3 / myocardial fibrosis

Fundings

This work was supported by the National Natural Science Foundation of China (Grant Nos. 82070234 and 82100254).

Acknowledgments

We appreciate Collaborative Innovation Center for Cardiovascular Disease Translational Medicine for technical support. We are gratefull to Yuehua Li laboratory members for technical help, reagents, and comments on the manuscript. The Graphical abstract was created with BioRender.com.

References

[1] Ramanujam D, Schön AP, Beck C, et al. MicroRNA-21-dependent macrophage-to-fibroblast signaling determines the cardiac response to pressure overload[J]. Circulation, 2021, 143(15): 1513-1525. doi: 10.1161/CIRCULATIONAHA.120.050682

[2] Barallobre-Barreiro J, Radovits T, Fava M, et al. Extracellular matrix in heart failure: role of ADAMTS5 in proteoglycan remodeling[J]. Circulation, 2021, 144(25): 2021-2034. doi: 10.1161/CIRCULATIONAHA.121.055732

[3] Zhang S, Li Y, Huang X, et al. Seamless genetic recording of transiently activated mesenchymal gene expression in endothelial cells during cardiac fibrosis[J]. Circulation, 2021, 144(25): 2004-2020. doi: 10.1161/CIRCULATIONAHA.121.055417

[4] Gourdie RG, Dimmeler S, Kohl P. Novel therapeutic strategies targeting fibroblasts and fibrosis in heart disease[J]. Nat Rev Drug Discov, 2016, 15(9): 620-638. doi: 10.1038/nrd.2016.89

[5] Frangogiannis NG. Transforming growth factor-β in myocardial disease[J]. Nat Rev Cardiol, 2022, 19(7): 435-455. doi: 10.1038/s41569-021-00646-w

[6] Blyszczuk P, Müller-Edenborn B, Valenta T, et al. Transforming growth factor-β-dependent Wnt secretion controls myofibroblast formation and myocardial fibrosis progression in experimental autoimmune myocarditis[J]. Eur Heart J, 2017, 38(18): 1413-1425. doi: 10.1093/eurheartj/ehw116

[7] Yan X, Lin Z, Chen F, et al. Human BAMBI cooperates with Smad7 to inhibit transforming growth factor-beta signaling[J]. J Biol Chem, 2009, 284(44): 30097-30104. doi: 10.1074/jbc.M109.049304

[8] Qi J, Yu Y, Öztürk ÖA, et al. New Wnt/β-catenin target genes promote experimental metastasis and migration of colorectal cancer cells through different signals[J]. Gut, 2016, 65(10): 1690-1701. doi: 10.1136/gutjnl-2014-307900

[9] Costa Gomes CP, Schroen B, Kuster GM, et al. Regulatory RNAs in heart failure[J]. Circulation, 2020, 141(4): 313-328. doi: 10.1161/CIRCULATIONAHA.119.042474

[10] Orso F, Quirico L, Dettori D, et al. Role of miRNAs in tumor and endothelial cell interactions during tumor progression[J]. Semin Cancer Biol, 2020, 60: 214-224. doi: 10.1016/j.semcancer.2019.07.024

[11] Carè A, Catalucci D, Felicetti F, et al. MicroRNA-133 controls cardiac hypertrophy[J]. Nat Med, 2007, 13(5): 613-618. doi: 10.1038/nm1582

[12] Zhao M, Chen N, Li X, et al. MiR-19a modulates hypoxia-mediated cell proliferation and migration via repressing PTEN in human pulmonary arterial smooth muscle[J]. Life Sci, 2019, 239: 116928. doi: 10.1016/j.lfs.2019.116928

[13] Xie C, Zhu J, Yang X, et al. TAp63α is involved in tobacco smoke-induced lung cancer EMT and the anti-cancer activity of curcumin via miR-19 transcriptional suppression[J]. Front Cell Dev Biol, 2021, 9: 645402. doi: 10.3389/fcell.2021.645402

[14] Song J, Zhu Y, Li J, et al. Pellino1-mediated TGF-β1 synthesis contributes to mechanical stress induced cardiac fibroblast activation[J]. J Mol Cell Cardiol, 2015, 79: 145-156. doi: 10.1016/j.yjmcc.2014.11.006

[15] Wu W, Hu Y, Li J, et al. Silencing of Pellino1 improves post-infarct cardiac dysfunction and attenuates left ventricular remodelling in mice[J]. Cardiovasc Res, 2014, 102(1): 46-55. doi: 10.1093/cvr/cvu007

[16] Liao X, Wang X, Jin H, et al. Mechanical stretch induces mitochondria-dependent apoptosis in neonatal rat cardiomyocytes and G₂/M accumulation in cardiac fibroblasts[J]. Cell Res, 2004, 14(1): 16-26. doi: 10.1038/sj.cr.7290198

[17] Zeng Z, Wang Q, Yang X, et al. Qishen granule attenuates cardiac fibrosis by regulating TGF-β /Smad3 and GSK-3β pathway[J]. Phytomedicine, 2019, 62: 152949. doi: 10.1016/j.phymed.2019.152949

[18] Verma SK, Garikipati VNS, Krishnamurthy P, et al. Interleukin-10 inhibits bone marrow fibroblast progenitor cell-mediated cardiac fibrosis in pressure-overloaded myocardium[J]. Circulation, 2017, 136(10): 940-953. doi: 10.1161/CIRCULATIONAHA.117.027889

[19] Weber KT, Sun Y, Bhattacharya SK, et al. Myofibroblast-mediated mechanisms of pathological remodelling of the heart[J]. Nat Rev Cardiol, 2013, 10(1): 15-26. doi: 10.1038/nrcardio.2012.158

[20] Batlle E, Massagué J. Transforming growth factor-β signaling in immunity and cancer[J]. Immunity, 2019, 50(4): 924-940. doi: 10.1016/j.immuni.2019.03.024

[21] Nagalingam RS, Chattopadhyaya S, Al-Hattab DS, et al. Scleraxis and fibrosis in the pressure-overloaded heart[J]. Eur Heart J, 2022, 43(45): 4739-4750. doi: 10.1093/eurheartj/ehac362

[22] Umbarkar P, Singh AP, Gupte M, et al. Cardiomyocyte SMAD4-dependent TGF-β signaling is essential to maintain adult heart homeostasis[J]. JACC Basic Transl Sci, 2019, 4(1): 41-53. doi: 10.1016/j.jacbts.2018.10.003

[23] Yao Y, Hu C, Song Q, et al. ADAMTS16 activates latent TGF-β accentuating fibrosis and dysfunction of the pressure-overloaded heart[J]. Cardiovasc Res, 2020, 116(5): 956-969. doi: 10.1093/cvr/cvz187

[24] Rodriguez P, Sassi Y, Troncone L, et al. Deletion of delta-like 1 homologue accelerates fibroblast-myofibroblast differentiation and induces myocardial fibrosis[J]. Eur Heart J, 2019, 40(12): 967-978. doi: 10.1093/eurheartj/ehy188

[25] Pchejetski D, Foussal C, Alfarano C, et al. Apelin prevents cardiac fibroblast activation and collagen production through inhibition of sphingosine kinase 1[J]. Eur Heart J, 2012, 33(18): 2360-2369. doi: 10.1093/eurheartj/ehr389

[26] Verjans R, Peters T, Beaumont FJ, et al. MicroRNA-221/222 family counteracts myocardial fibrosis in pressure overload-induced heart failure[J]. Hypertension, 2018, 71(2): 280-288. doi: 10.1161/HYPERTENSIONAHA.117.10094

[27] Jusic A, Thomas PB, Wettinger SB, et al. Noncoding RNAs in age-related cardiovascular diseases[J]. Ageing Res Rev, 2022, 77: 101610. doi: 10.1016/j.arr.2022.101610

[28] Zou M, Wang F, Gao R, et al. Autophagy inhibition of hsa-miR-19a-3p/19b-3p by targeting TGF-β R II during TGF-β1-induced fibrogenesis in human cardiac fibroblasts[J]. Sci Rep, 2016, 6: 24747. doi: 10.1038/srep24747

[29] Zhang Y, Zhang G, Che L, et al. miR-19 promotes development of renal fibrosis by targeting PTEN-mediated epithelial-mesenchymal transition[J]. Int J Clin Exp Pathol, 2020, 13(4): 642-654. https://pubmed.ncbi.nlm.nih.gov/32355512/

[30] Small EM, Olson EN. Pervasive roles of microRNAs in cardiovascular biology[J]. Nature, 2011, 469(7330): 336-342. doi: 10.1038/nature09783

[31] Zhang J, Chen G, Chen L, et al. TGF-β/BAMBI pathway dysfunction contributes to peripheral Th17/Treg imbalance in chronic obstructive pulmonary disease[J]. Sci Rep, 2016, 6: 31911. doi: 10.1038/srep31911

[32] Onichtchouk D, Chen Y, Dosch R, et al. Silencing of TGF-β signalling by the pseudoreceptor BAMBI[J]. Nature, 1999, 401(6752): 480-485. doi: 10.1038/46794

[33] Lee S, Lee MJ, Zhang J, et al. C-terminal-truncated HBV X promotes hepato-oncogenesis through inhibition of tumor-suppressive β-catenin/BAMBI signaling[J]. Exp Mol Med, 2016, 48(12): e275. doi: 10.1038/emm.2016.107

[34] Villar AV, García R, Llano M, et al. BAMBI (BMP and activin membrane-bound inhibitor) protects the murine heart from pressure-overload biomechanical stress by restraining TGF-β signaling[J]. Biochim Biophys Acta, 2013, 1832(2): 323-235. doi: 10.1016/j.bbadis.2012.11.007