Cardiomyocyte precursors generated by direct reprogramming and molecular beacon selection attenuate ventricular remodeling after experimental myocardial infarction

Stem Cell Research & Therapy

Research

Andreas Kurtz¹  Grit Nebrich²  Oliver Klein²  Kristin Klose³  Matthias Becker³  Dipthi Bachamanda Somesh³  Ulrike Krüger⁴  Karsten Jürchott⁴  Timo Z. Nazari-Shafti⁵  Volkmar Falk⁶  Christof Stamm⁷  Désirée Kunkel⁸  Manfred Gossen⁹  Janita A. Maring^1,10  Stephanie I. Protze^1,11 

[1] BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;BIH Imaging Mass Spectrometry Core Unit, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;Berlin-Brandenburg School for Regenerative Therapies, Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;Charité – Universitätsmedizin Berlin, Institute for Medical Immunology, 13353, Berlin, Germany;BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;Department of Cardiothoracic and Vascular Surgery, Deutsches Herzzentrum der Charité – Medical Heart Center of Charité and German Heart Institute Berlin, Augustenburger Platz 1, 13353, Berlin, Germany;German Centre for Cardiovascular Research, Partner Site Berlin, 10785, Berlin, Germany;BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;Department of Cardiothoracic and Vascular Surgery, Deutsches Herzzentrum der Charité – Medical Heart Center of Charité and German Heart Institute Berlin, Augustenburger Platz 1, 13353, Berlin, Germany;German Centre for Cardiovascular Research, Partner Site Berlin, 10785, Berlin, Germany;Department of Health Sciences and Technology, ETH Zurich, 8092, Zurich, Switzerland;BIH Center for Regenerative Therapies, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;Institute of Active Polymers, Helmholtz-Zentrum Hereon, 14513, Teltow, Germany;Berlin-Brandenburg Center for Regenerative Therapies, 13353, Berlin, Germany;Department of Cardiothoracic and Vascular Surgery, Deutsches Herzzentrum der Charité – Medical Heart Center of Charité and German Heart Institute Berlin, Augustenburger Platz 1, 13353, Berlin, Germany;German Centre for Cardiovascular Research, Partner Site Berlin, 10785, Berlin, Germany;Cytometry Core Facility, Berlin Institute of Health at Charité – Universitätsmedizin Berlin, 13353, Berlin, Germany;Institute of Active Polymers, Helmholtz-Zentrum Hereon, 14513, Teltow, Germany;Berlin-Brandenburg Center for Regenerative Therapies, 13353, Berlin, Germany;Institute of Active Polymers, Helmholtz-Zentrum Hereon, 14513, Teltow, Germany;Berlin-Brandenburg Center for Regenerative Therapies, 13353, Berlin, Germany;Department of Cardiothoracic and Vascular Surgery, Deutsches Herzzentrum der Charité – Medical Heart Center of Charité and German Heart Institute Berlin, Augustenburger Platz 1, 13353, Berlin, Germany;University Health Network, McEwen Stem Cell Institute, M5G 1L7, Toronto, ON, Canada;Department of Molecular Genetics, University of Toronto, M5S 1A8, Toronto, ON, Canada;

关键词: Cell therapy;    Direct reprogramming;    Fibroblast;    Induced cardiomyocyte precursor;    Molecular beacon;    Myocardial infarction;    Transdifferentiation;   

DOI  :  10.1186/s13287-023-03519-w

 received in 2022-03-24, accepted in 2023-09-25,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundDirect cardiac reprogramming is currently being investigated for the generation of cells with a true cardiomyocyte (CM) phenotype. Based on the original approach of cardiac transcription factor-induced reprogramming of fibroblasts into CM-like cells, various modifications of that strategy have been developed. However, they uniformly suffer from poor reprogramming efficacy and a lack of translational tools for target cell expansion and purification. Therefore, our group has developed a unique approach to generate proliferative cells with a pre-CM phenotype that can be expanded in vitro to yield substantial cell doses.MethodsCardiac fibroblasts were reprogrammed toward CM fate using lentiviral transduction of cardiac transcriptions factors (GATA4, MEF2C, TBX5, and MYOCD). The resulting cellular phenotype was analyzed by RNA sequencing and immunocytology. Live target cells were purified based on intracellular CM marker expression using molecular beacon technology and fluorescence-activated cell sorting. CM commitment was assessed using 5-azacytidine-based differentiation assays and the therapeutic effect was evaluated in a mouse model of acute myocardial infarction using echocardiography and histology. The cellular secretome was analyzed using mass spectrometry.ResultsWe found that proliferative CM precursor-like cells were part of the phenotype spectrum arising during direct reprogramming of fibroblasts toward CMs. These induced CM precursors (iCMPs) expressed CPC- and CM-specific proteins and were selectable via hairpin-shaped oligonucleotide hybridization probes targeting Myh6/7-mRNA–expressing cells. After purification, iCMPs were capable of extensive expansion, with preserved phenotype when under ascorbic acid supplementation, and gave rise to CM-like cells with organized sarcomeres in differentiation assays. When transplanted into infarcted mouse hearts, iCMPs prevented CM loss, attenuated fibrotic scarring, and preserved ventricular function, which can in part be attributed to their substantial secretion of factors with documented beneficial effect on cardiac repair.ConclusionsFibroblast reprogramming combined with molecular beacon-based cell selection yields an iCMP-like cell population with cardioprotective potential. Further studies are needed to elucidate mechanism-of-action and translational potential.Graphical Abstract

【 授权许可】

CC BY   
© BioMed Central Ltd., part of Springer Nature 2023

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