Generation of First Heart Field-like Cardiac Progenitors and Ventricular-like Cardiomyocytes from Human Pluripotent Stem Cells

Yu, Michael; Spiering, Sean; Colas, Alexandre R.

doi:10.3791/57688

Cited by 14 publications

(22 citation statements)

References 19 publications

(36 reference statements)

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“…Since PGC1/PPARα signals promoted CM contractility, we next sought to identify genes mediating the process. To do this, we selected 148 genes significantly downregulated in p7 PGC1 cmKO CMs (Table S2) and performed a single-cell highthroughput functional assay with PSC-CMs [36][37][38] treated with PPARα-specific ligands (Fig. 6a).…”

Section: Resultsmentioning

confidence: 99%

PGC1/PPAR drive cardiomyocyte maturation at single cell level via YAP1 and SF3B2

et al. 2021

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Cardiomyocytes undergo significant structural and functional changes after birth, and these fundamental processes are essential for the heart to pump blood to the growing body. However, due to the challenges of isolating single postnatal/adult myocytes, how individual newborn cardiomyocytes acquire multiple aspects of the mature phenotype remains poorly understood. Here we implement large-particle sorting and analyze single myocytes from neonatal to adult hearts. Early myocytes exhibit wide-ranging transcriptomic and size heterogeneity that is maintained until adulthood with a continuous transcriptomic shift. Gene regulatory network analysis followed by mosaic gene deletion reveals that peroxisome proliferator-activated receptor coactivator-1 signaling, which is active in vivo but inactive in pluripotent stem cell-derived cardiomyocytes, mediates the shift. This signaling simultaneously regulates key aspects of cardiomyocyte maturation through previously unrecognized proteins, including YAP1 and SF3B2. Our study provides a single-cell roadmap of heterogeneous transitions coupled to cellular features and identifies a multifaceted regulator controlling cardiomyocyte maturation.

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Section: Resultsmentioning

confidence: 99%

PGC1/PPAR drive cardiomyocyte maturation at single cell level via YAP1 and SF3B2

et al. 2021

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“…Animals and PSC Culture. PGC1α/β flox, Ai9, Yap1flox mice (Lai et al, 2008;Lin et al, 2004;Madisen et al, 2010;Zhang et al, 2010) were obtained from the Jackson Laboratory. All protocols involving animals followed U.S NIH guidelines and were approved by the animal and care use committee of the Johns Hopkins Medical Institutions.…”

Section: Methodsmentioning

confidence: 99%

“…Since PGC1/PPARα signals promoted CM contractility, we sought to identify downstream genes regulating calcium han-dling. To do this, we performed a single-cell high-throughput functional assay with PSC-CMs (Cunningham et al, 2017;McKeithan et al, 2017;Yu et al, 2018) treated with PPARαspecific ligands as shown in Figure 6A. Single cell analysis of calcium handling revealed that ligand-treated PSC-CMs have shorter ( 30ms) calcium transient duration (CTD) as compared to vehicle treated cells (DMSO) ( Figure 6B and Figure 6C).…”

Section: Pgc1/pparα Signaling Promotes CM Maturation By Regulating Kementioning

confidence: 99%

PGC1/PPAR Drive Cardiomyocyte Maturation through Regulation of Yap1 and SF3B2

Murphy

Miyamoto

Kervadec

et al. 2020

Preprint

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Cardiomyocytes undergo significant levels of structural and functional changes after birth-fundamental processes essential for the heart to produce the volume and contractility to pump blood to the growing body. However, due to the challenges in isolating single postnatal/adult myocytes, how individual newborn cardiomyocytes acquire multiple aspects of mature phenotypes remains poorly understood. Here we implemented large-particle sorting and analyzed single myocytes from neonatal to adult hearts. Early myocytes exhibited a wide-ranging transcriptomic and size heterogeneity, maintained until adulthood with a continuous transcriptomic shift. Gene regulatory network analysis followed by mosaic gene deletion revealed that peroxisome proliferator-activated receptor coactivator-1 signaling-activated in vivo but inactive in pluripotent stem cellderived cardiomyocytes-mediates the shift. The signaling regulated key aspects of cardiomyocyte maturation simultaneously through previously unrecognized regulators, including Yap1 and SF3B2. Our study provides a single-cell roadmap of heterogeneous transitions coupled to cellular features and unveils a multifaceted regulator controlling cardiomyocyte maturation. bioRxiv | February 6, 2020 1-16

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“…hiPSC-derived cardiomyocytes were produced as previously described. 23, 24 Cardiomyocytes were plated in 384 wells and transfected with siRNAs (see Online Appendix). Two days after transfection, EdU was added to the media for 24 hours and fixed.…”

Section: Methodsmentioning

confidence: 99%

Patient-specific functional genomics and disease modeling suggest a role for LRP2 in hypoplastic left heart syndrome

Jl¹,

Vogler

Missinato

et al. 2019

Preprint

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Congenital heart diseases (CHD), such as hypoplastic left heart syndrome (HLHS), are considered to have complex genetic underpinnings that are poorly understood. Here, an integrated multi-disciplinary approach was applied to identify novel genes and underlying mechanisms associated with HLHS. A family-based strategy was employed that coupled whole genome sequencing (WGS) with RNA sequencing of patient-derived induced pluripotent stem cells (iPSCs) from a sporadic HLHS proband-parent trio to identify, prioritize and functionally evaluate candidate genes in model systems. Consistent with the hypoplastic phenotype, the proband’s iPSCs had reduced proliferation capacity. Filtering WGS for rare de novo, recessive, and loss-of-function variants revealed 10 candidate genes with recessive variants and altered expression compared to the parents’ iPSCs. siRNA/RNAi-mediated knockdown in generic human iPSC-derived cardiac progenitors and in the in vivo Drosophila heart model revealed that LDL receptor related protein LRP2 and apolipoprotein APOB are required for robust hiPSC-derived cardiomyocyte proliferation and normal hear structure and function, possibly involving an oligogenic mechanism via growth-promoting WNT and SHH signaling. LRP2 was further validated as a CHD gene in a zebrafish heart model and rare variant burden testing in an HLHS cohort. Collectively, this cross-functional genetic approach to complex congenital heart disease revealed LRP2 dysfunction as a likely novel genetic driver of HLHS, and hereby established a scalable approach to decipher the oligogenic underpinnings of maladaptive left heart development.One sentence summaryWhole genome sequencing and a multi-model system candidate gene validation - human iPSC-derived cardiomyocytes and Drosophila and zebrafish hearts - identified lipoprotein LRP2 as a new potential driver in congenital heart disease and suggests a deficit in proliferation as a hallmark of hypoplastic left heart syndrome.

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Generation of First Heart Field-like Cardiac Progenitors and Ventricular-like Cardiomyocytes from Human Pluripotent Stem Cells

Cited by 14 publications

References 19 publications

PGC1/PPAR drive cardiomyocyte maturation at single cell level via YAP1 and SF3B2

PGC1/PPAR drive cardiomyocyte maturation at single cell level via YAP1 and SF3B2

PGC1/PPAR Drive Cardiomyocyte Maturation through Regulation of Yap1 and SF3B2

Patient-specific functional genomics and disease modeling suggest a role for LRP2 in hypoplastic left heart syndrome

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