{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T08:01:49Z","timestamp":1780128109720,"version":"3.54.0"},"reference-count":35,"publisher":"IMR Press","issue":"5","license":[{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100017610","name":"Shenzhen Science and Technology Innovation Program","doi-asserted-by":"crossref","award":["JCYJ20230807141801002"],"award-info":[{"award-number":["JCYJ20230807141801002"]}],"id":[{"id":"10.13039\/501100017610","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100017610","name":"Shenzhen Science and Technology Innovation Program","doi-asserted-by":"crossref","award":["JCYJ20240813160259020"],"award-info":[{"award-number":["JCYJ20240813160259020"]}],"id":[{"id":"10.13039\/501100017610","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Longgang District Science and Technology Innovation Bureau Project","award":["LGKCYLWS2022025"],"award-info":[{"award-number":["LGKCYLWS2022025"]}]},{"name":"Longgang District Key Specialty Program"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Front. Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background: The development of anti-fibrotic therapies for heart failure is hindered by the absence of phenotypically stable and scalable human cardiac fibroblast (CFB) models suitable for pre-clinical drug screening. This study aimed to establish and validate a conditionally immortalized CFB (iCFB) cell line for screening cardiac fibrosis drugs. Methods: An iCFB line was generated using a doxycycline (dox)-inducible system. The cellular characteristics were evaluated using molecular, protein, and functional analyses. The utility of the model was assessed by exposure to multiple pro-fibrotic stimuli (transforming growth factor-\u03b21 (TGF-\u03b21), angiotensin II, and palmitic acid) and two anti-fibrotic compounds (N-[(1R)-1,2,3,4-tetrahydro-1-naphthalenyl]-1H-benzimidazol-2-amine [NS8593] and pirfenidone). Results: iCFB proliferation was tightly regulated by dox. Upon dox withdrawal, the iCFBs reverted to a quiescent state and exhibited a molecular expression profile (collagen type I alpha 1 chain [COL1A1], periostin [POSTN], gap junction protein alpha 1 [GJA1], and T-box transcription factor 20 [TBX20]) comparable to that of primary CFBs. This phenotypic fidelity, along with a robust capacity for TGF-\u03b21-induced myofibroblast differentiation, was maintained during long-term culture up to population doubling 60. The model responded consistently to diverse pro-fibrotic stimuli, confirming the anti-fibrotic efficacy of the transient receptor potential melastatin 7 (TRPM7) inhibitor, NS8593, and demonstrating the therapeutic potential of the repurposed drug pirfenidone. Conclusions: We developed a novel iCFB model that integrates long-term expandability, high biological fidelity, and broad responsiveness to fibrotic signaling. This robust platform is well-suited for mechanistic studies and drug screening, thereby facilitating and accelerating the discovery of anti-fibrotic therapeutics.<\/jats:p>","DOI":"10.31083\/fbl49465","type":"journal-article","created":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T07:25:36Z","timestamp":1780125936000},"source":"Crossref","is-referenced-by-count":0,"title":["Conditional Immortalization of Human Cardiac Fibroblasts for Pro-Fibrotic and Anti-Fibrotic Drug Screening"],"prefix":"10.31083","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-6059-0992","authenticated-orcid":true,"given":"Sushan","family":"Wang","sequence":"first","affiliation":[{"name":"School of Life Sciences, Anhui Medical University, 230601 Hefei, Anhui, China"},{"name":"Central Laboratory, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1194-8078","authenticated-orcid":true,"given":"Ruijuan","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Cardiology, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9127-6323","authenticated-orcid":true,"given":"Rui","family":"Guo","sequence":"additional","affiliation":[{"name":"Central Laboratory, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-3646-4381","authenticated-orcid":true,"given":"Cheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Cardiology, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6905-599X","authenticated-orcid":true,"given":"Wenan","family":"Kang","sequence":"additional","affiliation":[{"name":"Central Laboratory, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0909-7710","authenticated-orcid":true,"given":"Jia","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Anhui Medical University, 230601 Hefei, Anhui, China"},{"name":"Central Laboratory, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-2661-2405","authenticated-orcid":true,"given":"Canling","family":"Long","sequence":"additional","affiliation":[{"name":"Central Laboratory, School of Medicine, The Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen & Longgang District People\u2019s Hospital of Shenzhen, 518172 Shenzhen, Guangdong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,4,30]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Tallquist MD. Cardiac Fibroblast Diversity. Annual Review of Physiology. 2020; 82: 63\u201378.","DOI":"10.1146\/annurev-physiol-021119-034527"},{"key":"ref2","doi-asserted-by":"publisher","unstructured":"Souders CA, Bowers SLK, Baudino TA. Cardiac fibroblast: the renaissance cell. Circulation Research. 2009; 105: 1164\u20131176.","DOI":"10.1161\/CIRCRESAHA.109.209809"},{"key":"ref3","doi-asserted-by":"publisher","unstructured":"Dostal D, Glaser S, Baudino TA. Cardiac fibroblast physiology and pathology. Comprehensive Physiology. 2015; 5: 887\u2013909.","DOI":"10.1002\/cphy.c140053"},{"key":"ref4","doi-asserted-by":"publisher","unstructured":"Gourdie RG, Dimmeler S, Kohl P. Novel therapeutic strategies targeting fibroblasts and fibrosis in heart disease. Nature Reviews. Drug Discovery. 2016; 15: 620\u2013638.","DOI":"10.1038\/nrd.2016.89"},{"key":"ref5","doi-asserted-by":"publisher","unstructured":"Weber KT, Sun Y, Bhattacharya SK, Ahokas RA, Gerling IC. Myofibroblast-mediated mechanisms of pathological remodelling of the heart. Nature Reviews. Cardiology. 2013; 10: 15\u201326.","DOI":"10.1038\/nrcardio.2012.158"},{"key":"ref6","doi-asserted-by":"publisher","unstructured":"Porter KE, Turner NA. Cardiac fibroblasts: at the heart of myocardial remodeling. Pharmacology & Therapeutics. 2009; 123: 255\u2013278.","DOI":"10.1016\/j.pharmthera.2009.05.002"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"Frangogiannis NG. Cardiac fibrosis: Cell biological mechanisms, molecular pathways and therapeutic opportunities. Molecular Aspects of Medicine. 2019; 65: 70\u201399.","DOI":"10.1016\/j.mam.2018.07.001"},{"key":"ref8","doi-asserted-by":"publisher","unstructured":"Lunde IG, Rypdal KB, Van Linthout S, Diez J, Gonz\u00e1lez A. Myocardial fibrosis from the perspective of the extracellular matrix: Mechanisms to clinical impact. Matrix Biology. 2024; 134: 1\u201322.","DOI":"10.1016\/j.matbio.2024.08.008"},{"key":"ref9","doi-asserted-by":"publisher","unstructured":"Leask A. Getting to the heart of the matter: new insights into cardiac fibrosis. Circulation Research. 2015; 116: 1269\u20131276.","DOI":"10.1161\/CIRCRESAHA.116.305381"},{"key":"ref10","doi-asserted-by":"publisher","unstructured":"Burke RM, Burgos Villar KN, Small EM. Fibroblast contributions to ischemic cardiac remodeling. Cellular Signalling. 2021; 77: 109824.","DOI":"10.1016\/j.cellsig.2020.109824"},{"key":"ref11","doi-asserted-by":"publisher","unstructured":"Ishiura J, Nakamori S, Ishida M, Dohi K. \u2018Targeting the cardiac myocyte and fibrosis\u2019 in heart failure. European Heart Journal. 2022; 43: 432.","DOI":"10.1093\/eurheartj\/ehab780"},{"key":"ref12","doi-asserted-by":"publisher","unstructured":"L\u00f3pez B, Ravassa S, Moreno MU, Jos\u00e9 GS, Beaumont J, Gonz\u00e1lez A, et al. Diffuse myocardial fibrosis: mechanisms, diagnosis and therapeutic approaches. Nature Reviews. Cardiology. 2021; 18: 479\u2013498.","DOI":"10.1038\/s41569-020-00504-1"},{"key":"ref13","doi-asserted-by":"publisher","unstructured":"Chimenti I, Pagano F, Cozzolino C, Icolaro F, Floris E, Picchio V. The Role of Cardiac Fibroblast Heterogeneity in Myocardial Fibrosis and Its Novel Therapeutic Potential. International Journal of Molecular Sciences. 2025; 26: 5882.","DOI":"10.3390\/ijms26125882"},{"key":"ref14","doi-asserted-by":"publisher","unstructured":"Cinato M, Kang R, Kramar S, Savchenko L, Pizzinat N, Swiader A, et al. Transcriptome fingerprinting of aberrant fibroblast activation unlocks effective therapeutics to tackle cardiac fibrosis. Science Advances. 2025; 11: eadx0968.","DOI":"10.1126\/sciadv.adx0968"},{"key":"ref15","doi-asserted-by":"publisher","unstructured":"Meng XY, Li Y , Meng LB, Yang CG, Xia CX, Wang X, et al. Growth Differentiation Factor 15 Inhibits Cardiac Fibrosis, Oxidative Stress, Inflammation, and Apoptosis in a Rat Model of Heart Failure with Preserved Ejection Fraction. Front. Biosci. (Landmark Ed). 2025; 30: eadx26857.","DOI":"10.31083\/FBL26857"},{"key":"ref16","doi-asserted-by":"publisher","unstructured":"Fang L, Murphy AJ, Dart AM. A Clinical Perspective of Anti-Fibrotic Therapies for Cardiovascular Disease. Frontiers in Pharmacology. 2017; 8: 186.","DOI":"10.3389\/fphar.2017.00186"},{"key":"ref17","doi-asserted-by":"publisher","unstructured":"Spoladore R, Falasconi G, Fiore G, Di Maio S, Preda A, Slavich M, et al. Cardiac fibrosis: emerging agents in preclinical and clinical development. Expert Opinion on Investigational Drugs. 2021; 30: 153\u2013166.","DOI":"10.1080\/13543784.2021.1868432"},{"key":"ref18","doi-asserted-by":"publisher","unstructured":"Zhao M, Wang L, Wang M, Zhou S, Lu Y, Cui H, et al. Targeting fibrosis, mechanisms and cilinical trials. Signal Transduction and Targeted Therapy. 2022; 7: 206.","DOI":"10.1038\/s41392-022-01070-3"},{"key":"ref19","doi-asserted-by":"publisher","unstructured":"Kel A, Thum T, Kunduzova O. Targeting fibroblast phenotype switching in cardiac remodelling as a promising antifibrotic strategy. European Heart Journal. 2025; 46: 354\u2013358.","DOI":"10.1093\/eurheartj\/ehae722"},{"key":"ref20","doi-asserted-by":"publisher","unstructured":"Xu Q, Norman JT, Shrivastav S, Lucio-Cazana J, Kopp JB. In vitro models of TGF-beta-induced fibrosis suitable for high-throughput screening of antifibrotic agents. American Journal of Physiology. Renal Physiology. 2007; 293: F631\u2013F640.","DOI":"10.1152\/ajprenal.00379.2006"},{"key":"ref21","doi-asserted-by":"publisher","unstructured":"Liao G, Long C, Guo R, Wang S, Han R, Yang J, et al. Conditional immortalization of mesenchymal stem cells and their extracellular vesicles therapy for interstitial cystitis\/bladder pain syndrome. Stem Cell Research & Therapy. 2025; 16: 459.","DOI":"10.1186\/s13287-025-04615-9"},{"key":"ref22","doi-asserted-by":"publisher","unstructured":"Liu J, Volkers L, Jangsangthong W, Bart CI, Engels MC, Zhou G, et al. Generation and primary characterization of iAM-1, a versatile new line of conditionally immortalized atrial myocytes with preserved cardiomyogenic differentiation capacity. Cardiovascular Research. 2018; 114: 1848\u20131859.","DOI":"10.1093\/cvr\/cvy134"},{"key":"ref23","doi-asserted-by":"publisher","unstructured":"Jim\u00e9nez-Gonz\u00e1lez S, Mar\u00edn-Royo G, Jurado-L\u00f3pez R, Bartolom\u00e9 MV, Romero-Miranda A, Luaces M, et al. The Crosstalk between Cardiac Lipotoxicity and Mitochondrial Oxidative Stress in the Cardiac Alterations in Diet-Induced Obesity in Rats. Cells. 2020; 9: 451.","DOI":"10.3390\/cells9020451"},{"key":"ref24","doi-asserted-by":"publisher","unstructured":"Bujak M, Frangogiannis NG. The role of TGF-beta signaling in myocardial infarction and cardiac remodeling. Cardiovascular Research. 2007; 74: 184\u2013195.","DOI":"10.1016\/j.cardiores.2006.10.002"},{"key":"ref25","doi-asserted-by":"publisher","unstructured":"Jana S, Aujla P, Hu M, Kilic T, Zhabyeyev P, McCulloch CA, et al. Gelsolin is an important mediator of Angiotensin II-induced activation of cardiac fibroblasts and fibrosis. FASEB Journal. 2021; 35: e21932.","DOI":"10.1096\/fj.202100038RR"},{"key":"ref26","doi-asserted-by":"publisher","unstructured":"Maruyama K, Imanaka-Yoshida K. The Pathogenesis of Cardiac Fibrosis: A Review of Recent Progress. International Journal of Molecular Sciences. 2022; 23: 2617.","DOI":"10.3390\/ijms23052617"},{"key":"ref27","doi-asserted-by":"publisher","unstructured":"Bacmeister L, Schwarzl M, Warnke S, Stoffers B, Blankenberg S, Westermann D, et al. Inflammation and fibrosis in murine models of heart failure. Basic Research in Cardiology. 2019; 114: 19.","DOI":"10.1007\/s00395-019-0722-5"},{"key":"ref28","doi-asserted-by":"publisher","unstructured":"Chubanov V, Gudermann T. Mapping TRPM7 Function by NS8593. International Journal of Molecular Sciences. 2020; 21: 7017.","DOI":"10.3390\/ijms21197017"},{"key":"ref29","doi-asserted-by":"publisher","unstructured":"Rios FJ, Zou ZG, Harvey AP, Harvey KY, Nosalski R, Anyfanti P, et al. Chanzyme TRPM7 protects against cardiovascular inflammation and fibrosis. Cardiovascular Research. 2020; 116: 721\u2013735.","DOI":"10.1093\/cvr\/cvz164"},{"key":"ref30","doi-asserted-by":"publisher","unstructured":"Suzuki S, Penner R, Fleig A. TRPM7 contributes to progressive nephropathy. Scientific Reports. 2020; 10: 2333.","DOI":"10.1038\/s41598-020-59355-y"},{"key":"ref31","doi-asserted-by":"publisher","unstructured":"Hu F, Li M, Han F, Zhang Q, Zeng Y, Zhang W, et al. Role of TRPM7 in cardiac fibrosis: A potential therapeutic target (Review). Experimental and Therapeutic Medicine. 2021; 21: 173.","DOI":"10.3892\/etm.2020.9604"},{"key":"ref32","doi-asserted-by":"publisher","unstructured":"Li N, Hang W, Shu H, Zhou N. Pirfenidone alleviates cardiac fibrosis induced by pressure overload via inhibiting TGF-\u03b21\/Smad3 signalling pathway. Journal of Cellular and Molecular Medicine. 2022; 26: 4548\u20134555.","DOI":"10.1111\/jcmm.17478"},{"key":"ref33","doi-asserted-by":"publisher","unstructured":"Aimo A, Spitaleri G, Panichella G, Lup\u00f3n J, Emdin M, Bayes-Genis A. Pirfenidone as a novel cardiac protective treatment. Heart Failure Reviews. 2022; 27: 525\u2013532.","DOI":"10.1007\/s10741-021-10175-w"},{"key":"ref34","doi-asserted-by":"publisher","unstructured":"Henderson NC, Rieder F, Wynn TA. Fibrosis: from mechanisms to medicines. Nature. 2020; 587: 555\u2013566.","DOI":"10.1038\/s41586-020-2938-9"},{"key":"ref35","doi-asserted-by":"publisher","unstructured":"King TE Jr, Bradford WZ, Castro-Bernardini S, Fagan EA, Glaspole I, Glassberg MK, et al. A phase 3 trial of pirfenidone in patients with idiopathic pulmonary fibrosis. The New England Journal of Medicine. 2014; 370: 2083\u20132092.","DOI":"10.1056\/NEJMoa1402582"}],"container-title":["Frontiers in Bioscience-Landmark"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.imrpress.com\/journal\/FBL\/31\/5\/10.31083\/FBL49465","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T07:26:52Z","timestamp":1780126012000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.imrpress.com\/journal\/FBL\/31\/5\/10.31083\/FBL49465"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,30]]},"references-count":35,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2026]]}},"alternative-id":["S2768-6701(26)02082-4"],"URL":"https:\/\/doi.org\/10.31083\/fbl49465","relation":{},"ISSN":["2768-6701","2768-6698"],"issn-type":[{"value":"2768-6701","type":"print"},{"value":"2768-6698","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,30]]},"article-number":"49465"}}