Natural Cardiogenesis-Based Template Predicts Cardiogenic Potential of Induced Pluripotent Stem Cell Lines

Abstract

Background Cardiac development is a complex process resulting in an integrated, multi-lineage tissue with developmental corruption in early embryogenesis leading to congenital heart disease. Interrogation of individual genes has provided the backbone for cardiac developmental biology, yet a comprehensive transcriptome derived from natural cardiogenesis is required to gauge innate developmental milestones. Methods and Results Stage-specific cardiac structures were dissected from eight distinctive mouse embryonic time points to produce genome-wide expressome analysis across cardiogenesis. In reference to this native cardiogenic expression roadmap, divergent iPSC-derived cardiac expression profiles were mapped from pro-cardiogenic 3-factor (SOX2, OCT4, KLF4) and less-cardiogenic 4-factor (plus c-MYC) reprogrammed cells. Expression of cardiac-related genes from 3F-iPSC differentiated in vitro at days 5 and 11 recapitulated expression profiles of natural embryos at days E7.5–E8.5 and E14.5–E18.5, respectively. In contrast, 4F-iPSC demonstrated incomplete cardiogenic gene expression profiles beginning at day 5 of differentiation. Differential gene expression within the pluripotent state revealed 23 distinguishing candidate genes among pluripotent cell lines with divergent cardiogenic potentials. A confirmed panel of 12 genes, differentially expressed between high and low cardiogenic lines, was transformed into a predictive score sufficient to discriminate individual iPSC lines according to relative cardiogenic potential. Conclusions Transcriptome analysis attuned to natural embryonic cardiogenesis provides a robust platform to probe coordinated cardiac specification and maturation from bioengineered stem cell-based model systems. A panel of developmental-related genes allowed differential prognosis of cardiogenic competency, thus prioritizing cell lines according to natural blueprint to streamline functional applications.

Identification of cardiovascular lineage descendants at single-cell resolution

“…Although ESC/iPSC-derived CMs may hold significant promise for cell-based therapy and as in vitro platforms for disease pathway and therapeutic drug discovery, their phenotype has been recognized as largely immature (Feinberg et al, 2013;Lieu et al, 2009;Liu et al, 2007;Martinez-Fernandez et al, 2013;Pillekamp et al, 2012;Robertson et al, 2013;Satin et al, 2008;Snir et al, 2003;Yang et al, 2014). Using our single-cell multiplex PCR expression analysis, we compared stage-by-stage the expression of main cardiac sarcomeric genes and transcription factors at distinct stages of development.…”

Role of regeneration in tissue repairing and therapies

“…Hence, injectable hyaluronic acid/adipic acid dihydrazide (HA/ADH) hydrogel an adiposetissue-like material was prepared that showed biological compatibility as well as stiff mechanical behavior upon use. This also supports growth, proliferation and differentiation of 3T3-L1 pre-adipocytes and serves as a vessel for protecting pre-adipocytes for short-term after delivery to native tissues [155]. Adipose tissue is a weight-bearing biological structure that needs many biological molecules for explant growth.…”

Induced Pluripotent Stem Cells for Cardiovascular Disease: From Product-Focused Disease Modeling to Process-Focused Disease Discovery

“…Toward the goal of defining the molecular timeline of heart development, recent studies describe a transcriptional atlas of normal cardiogenesis from the pluripotent state to adult cardiomyocytes [54, 55]. In these studies, authors map the dynamic transcriptional landscape of murine heart development from an unbiased genomic perspective.…”

“…While recent work has highlighted factors such as mechanical force that can induce sarcomeric maturity within iPS cell-derived cardiomyocytes [64], much more research is needed to consistently generate adult-like cardiomyocytes in vitro . To advance process-focused disease discovery applications, additional studies will be required to determine the accuracy with which iPS cell-derived cardiac progenitors align with natural progenitor stages during in utero heart development [55]. …”