P53 Activation Promotes Maturational Characteristics of Pluripotent Stem Cell-Derived Cardiomyocytes in 3-Dimensional Suspension Culture Via FOXO-FOXM1 Regulation.

Autor:	Velayutham N; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Garbern JC; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA.; Department of Cardiology Boston Children's Hospital Boston MA USA., Elwell HLT; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Zhuo Z; Bioinformatics Core, Department of Biostatistics Harvard T.H. Chan School of Public Health Boston MA USA., Rüland L; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Elcure Alvarez F; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Frontini S; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Rodriguez Carreras Y; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Eichholtz M; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Ricci-Blair E; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Shaw JY; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Bouffard AH; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Sokol M; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Mancheño Juncosa E; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Rhoades S; Ginkgo Bioworks Boston MA USA., van den Berg D; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Kreymerman A; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Aoyama J; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA., Höfflin J; Ginkgo Bioworks Boston MA USA., Ryan H; Ginkgo Bioworks Boston MA USA., Ho Sui S; Bioinformatics Core, Department of Biostatistics Harvard T.H. Chan School of Public Health Boston MA USA., Lee RT; Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute Harvard University Cambridge MA USA.; Division of Cardiovascular Medicine, Department of Medicine Brigham and Women's Hospital and Harvard Medical School Boston MA USA.
Jazyk:	angličtina
Zdroj:	Journal of the American Heart Association [J Am Heart Assoc] 2024 Jul 02; Vol. 13 (13), pp. e033155. Date of Electronic Publication: 2024 Jun 27.
DOI:	10.1161/JAHA.123.033155
Abstrakt:	Background: Current protocols generate highly pure human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in vitro that recapitulate characteristics of mature in vivo cardiomyocytes. Yet, a risk of arrhythmias exists when hiPSC-CMs are injected into large animal models. Thus, understanding hiPSC-CM maturational mechanisms is crucial for clinical translation. Forkhead box (FOX) transcription factors regulate postnatal cardiomyocyte maturation through a balance between FOXO and FOXM1. We also previously demonstrated that p53 activation enhances hiPSC-CM maturation. Here, we investigate whether p53 activation modulates the FOXO/FOXM1 balance to promote hiPSC-CM maturation in 3-dimensional suspension culture. Methods and Results: Three-dimensional cultures of hiPSC-CMs were treated with Nutlin-3a (p53 activator, 10 μM), LOM612 (FOXO relocator, 5 μM), AS1842856 (FOXO inhibitor, 1 μM), or RCM-1 (FOXM1 inhibitor, 1 μM), starting 2 days after onset of beating, with dimethyl sulfoxide (0.2% vehicle) as control. P53 activation promoted hiPSC-CM metabolic and electrophysiological maturation alongside FOXO upregulation and FOXM1 downregulation, in n=3 to 6 per group for all assays. FOXO inhibition significantly decreased expression of cardiac-specific markers such as TNNT2. In contrast, FOXO activation or FOXM1 inhibition promoted maturational characteristics such as increased contractility, oxygen consumption, and voltage peak maximum upstroke velocity, in n=3 to 6 per group for all assays. Further, by single-cell RNA sequencing of n=2 LOM612-treated cells compared with dimethyl sulfoxide, LOM612-mediated FOXO activation promoted expression of cardiac maturational pathways. Conclusions: We show that p53 activation promotes FOXO and suppresses FOXM1 during 3-dimensional hiPSC-CM maturation. These results expand our understanding of hiPSC-CM maturational mechanisms in a clinically-relevant 3-dimensional culture system.
Databáze:	MEDLINE
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