DAND5 Inactivation Enhances Cardiac Differentiation in Mouse Embryonic Stem Cells

Abstract

Deciphering the clues of a regenerative mechanism for the mammalian adult heart would save millions of lives in the near future. Heart failure due to cardiomyocyte loss is still one of the significant health burdens worldwide. Here, we show the potential of a single molecule, DAND5, in mouse pluripotent stem cell-derived cardiomyocytes specification and proliferation. Dand5 loss-of-function generated the double of cardiac beating foci compared to the wild-type cells. The early formation of cardiac progenitor cells and the increased proliferative capacity of Dand5 KO mESC-derived cardiomyocytes contribute to the observed higher number of derived cardiac cells. Transcriptional profiling sequencing and quantitative RT-PCR assays showed an upregulation of early cardiac gene networks governing cardiomyocyte differentiation, cell cycling, and cardiac regenerative pathways but reduced levels of genes involved in cardiomyocyte maturation. These findings prompt DAND5 as a key driver for the generation and expansion of pluripotent stem cell-derived cardiomyocytes systems with further clinical application purposes.

Gene-Edited Human-Induced Pluripotent Stem Cell Lines to Elucidate DAND5 Function throughout Cardiac Differentiation

“…As previously mentioned, a former study performed on mouse embryonic stem cells, has demonstrated that the suppression of DAND5 leads to an increase in proliferation of cardiomyocytes and drives to a distinct transcriptional profiling sequencing during the first stages of life [21]. In line with this study, we performed an extended structural and molecular analysis of DAND5-KO hiPSC-CMs.…”

“…Mutations in DAND5, such as the heterozygous c.455G>A missense mutation, which decreases DAND5 activity, and the homozygous truncating c.396_397dupCT variant predicted to produce a frameshift protein, have been strongly associated with clinically diagnosed heart malformations [14,22]. In mouse models, it was noticed that, among other cardiac defects, Dand5 loss-of-function promotes the proliferation of cardiomyocytes [18,21]. The hearts of Dand5 knockout mice display ventricular hypertrophy with increased ventricular wall thickness, which causes severe cardiac dysfunction [18].…”

“…The hearts of Dand5 knockout mice display ventricular hypertrophy with increased ventricular wall thickness, which causes severe cardiac dysfunction [18]. Moreover, using mouse embryonic stem cells (mESCs), it was shown that Dand5 loss-of-function dramatically increased the number of FLK-1+/PDGFR-α+ cardiac progenitor cells during mESCs cardiac differentiation [21]. In addition, the knockout of Dand5 seems to activate the transcription of several cell-cycle regulators, prompting cardiomyocyte proliferation [18,21].…”

“…Based on distinct mouse model studies, it becomes evident that DAND5 is associated with gene networks controlling differentiation and cell cycle of cardiomyocytes at early developmental stages [18,21]. The inactivation of Dand5 in cardiac populations increases the proliferative capacity of cardiomyocytes during mouse cardiac development [18] and mouse ES-CM differentiation [21].…”

“…Moreover, DAND5 was assigned as a molecule involved in cardiomyocyte proliferation/differentiation. Reduced DAND5 levels lead to prolonged TGF-β/Nodal and Wnt/β-catenin signalling during mouse embryonic stem cell cardiomyocyte differentiation, increasing cardiovascular progenitors' number and extending their progenitor state [21].…”