Abstract
Embryonic Stem Cells (ESCs) hold great potential for regeneration of damaged myocardium, however the molecular circuitry that guides ESC differentiation into cardiomyocytes remains poorly understood. This is exemplified by the elusive role of the transcription factor, Foxc1, during cardiac development. The only known Foxc1 target during heart development is Tbx1. Because Foxc1 null mice contain heart mutations that are far more severe than Tbx1 null mice, it is likely that Foxc1 has additional regulatory roles during heart development. The goal of our study was to test whether Foxc1 is critical for ESC differentiation into functional cardiomyocytes through proper regulation of specific downstream gene networks. Converging evidence from Foxc1 deficient and overexpression ESC models reveals a close relationship between Foxc1 levels and early cardiomyogenic factors Isl1, Mef2c, and Nkx2.5 and also the production of functional cardiomyocytes. We show Foxc1 regulates early cardiomyogenesis during a specific window of differentiation, D4-D6. Through whole transcriptome RNA-sequencing analysis, we report pathways regulated by Foxc1 involved in cardiac function including actin cytoskeleton, cell adhesion, tight and gap junctions, and calcium signaling. Our data indicate a novel Foxc1 direct gene target, Myh7, which encodes the predominant myosin heavy chain isoform, MHCβ, expressed during cardiac development. These data lead us to conclude that Foxc1 regulates both early cardiomyogenesis and the functional properties of ESC-derived cardiomyocytes. Our findings shed light on the molecular circuitry governing cardiomyogenesis that may lead to the development of better translational strategies for the use of pluripotent stem cells in regenerative medicine towards repairing damaged myocardium.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1487-1500 |
| Number of pages | 14 |
| Journal | Stem Cells |
| Volume | 34 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 1 2016 |
Funding
The transmission electron microscopy work was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. Lennell Reynolds provided imaging assistance. RNA- seq was performed at the University of Chicago genomics facility with support from Dr. Pieter Faber. RNA- seq pathway analysis was performed by Dr. Matt Schipma at the Next Generation Sequencing Core at Northwestern University. This work was supported in part by the National Institute of Health grants R01 HL093439 and R01 HL113541 (to G.Q.), R01 EY019484 and RO1 HL126920 (to T.K.), and R25 GM079300 (to E.L.) and American Heart Association's pre- doctoral fellowship grant 15PRE25080006 (to E.L.).
Keywords
- Beta
- Cardiomyocyte function
- Cardiomyogenesis
- Differentiation
- Embryonic stem cells
- Forkhead box C1
- Myosin heavy chain
ASJC Scopus subject areas
- Molecular Medicine
- Cell Biology
- Developmental Biology