Tumor Suppressors RB1 and CDKN2a Cooperatively Regulate Cell-Cycle Progression and Differentiation During Cardiomyocyte Development and Repair

Konstantinos E. Hatzistergos, Adam R. Williams, Derek M Dykxhoorn, Michael A. Bellio, Wendou Yu, Joshua Hare

Research output: Contribution to journalArticle

Abstract

RATIONALE: Although rare cardiomyogenesis is reported in the adult mammalian heart, whether this results from differentiation or proliferation of cardiomyogenic cells remains controversial. The tumor suppressor genes RB1 (retinoblastoma) and CDKN2a (cyclin-dependent kinase inhibitor 2a) are critical cell-cycle regulators, but their roles in human cardiomyogenesis remains unclear. OBJECTIVE: We hypothesized that developmental activation of RB1 and CDKN2a cooperatively cause permanent cell-cycle withdrawal of human cardiac precursors (CPCs) driving terminal differentiation into mature cardiomyocytes, and that dual inactivation of these tumor suppressor genes promotes myocyte cell-cycle reentry. METHODS AND RESULTS: Directed differentiation of human pluripotent stem cells (hPSCs) into cardiomyocytes revealed that RB1 and CDKN2a are upregulated at the onset of cardiac precursor specification, simultaneously with GATA4 (GATA-binding protein 4) homeobox genes PBX1 (pre-B-cell leukemia transcription factor 1) and MEIS1 (myeloid ecotropic viral integration site 1 homolog), and remain so until terminal cardiomyocyte differentiation. In both GATA4+ hPSC cardiac precursors and postmitotic hPSC-cardiomyocytes, RB1 is hyperphosphorylated and inactivated. Transient, stage-specific, depletion of RB1 during hPSC differentiation enhances cardiomyogenesis at the cardiac precursors stage, but not in terminally differentiated hPSC-cardiomyocytes, by transiently upregulating GATA4 expression through a cell-cycle regulatory pathway involving CDKN2a. Importantly, cytokinesis in postmitotic hPSC-cardiomyocytes can be induced with transient, dual RB1, and CDKN2a silencing. The relevance of this pathway in vivo was suggested by findings in a porcine model of cardiac cell therapy post-MI, whereby dual RB1 and CDKN2a inactivation in adult GATA4+ cells correlates with the degree of scar size reduction and endogenous cardiomyocyte mitosis, particularly in response to combined transendocardial injection of adult human hMSCs (bone marrow-derived mesenchymal stromal cells) and cKit+ cardiac cells. CONCLUSIONS: Together these findings reveal an important and coordinated role for RB1 and CDKN2a in regulating cell-cycle progression and differentiation during human cardiomyogenesis. Moreover, transient, dual inactivation of RB1 and CDKN2a in endogenous adult GATA4+ cells and cardiomyocytes mediates, at least in part, the beneficial effects of cell-based therapy in a post-MI large mammalian model, a finding with potential clinical implications.

Original languageEnglish (US)
Pages (from-to)1184-1197
Number of pages14
JournalCirculation research
Volume124
Issue number8
DOIs
StatePublished - Apr 12 2019

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Cyclin-Dependent Kinase Inhibitor p16
Cardiac Myocytes
Pluripotent Stem Cells
Cell Differentiation
Cell Cycle
Neoplasms
Carrier Proteins
Cell- and Tissue-Based Therapy
Tumor Suppressor Genes
Precursor B-Cell Lymphoblastic Leukemia-Lymphoma
Virus Integration
Cytokinesis
Retinoblastoma
Homeobox Genes
Mesenchymal Stromal Cells
Mitosis
Muscle Cells
Cicatrix
Transcription Factors
Swine

Keywords

  • cell cycle
  • cell proliferation
  • cytokinesis
  • mesenchymal stem cells
  • retinoblastoma protein

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Tumor Suppressors RB1 and CDKN2a Cooperatively Regulate Cell-Cycle Progression and Differentiation During Cardiomyocyte Development and Repair. / Hatzistergos, Konstantinos E.; Williams, Adam R.; Dykxhoorn, Derek M; Bellio, Michael A.; Yu, Wendou; Hare, Joshua.

In: Circulation research, Vol. 124, No. 8, 12.04.2019, p. 1184-1197.

Research output: Contribution to journalArticle

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AU - Williams, Adam R.

AU - Dykxhoorn, Derek M

AU - Bellio, Michael A.

AU - Yu, Wendou

AU - Hare, Joshua

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