It is now possible to direct the differentiation of pluripotent human stem cells into cardiomyocytes at a scale suitable for large experiments or small clinical trials. Despite this success, there are large gaps in our understanding of how signaling pathways converge on the nucleus to control cardiac differentiation. Furthermore, stem cell-derived cardiomyocytes are currently at a fetal stage of maturity, which may be poorly suited for modeling adult diseases or tissue repair. Project 3 addresses these issues with the following aims.
In Aim 1, we explore the biology of three candidate novel regulators of heart development identified by ChlPseq experiments. These include two transcription factors (MEIS2 and H0XB2) and a novel Wnt/b-catenin regulating protein (TMEM88). The genes will be knocked down in differentiating hESCs and in developing zebrafish embryos to determine their roles in cardiac differentiation and morphogenesis.
In Aim 2 we will utilize two lines of genetically modified hESCs that activate fluorescent protein expression when they differentiate to mesoderm (BRACHYURY knock-in) or first heart field cells (TBX5 knock-in). We will determine how their fates become restricted during differentiation in 2D and 3D culture and after transplantation in vivo. Additionally, we will use a Wnt/b-catenin reporter to track how this pathway positively and negatively regulates formation of these key cell types.
Aim 3 addresses the issue of cardiomyocyte maturation, beginning by exploring the role of 4 candidate miRNAs that are up-regulated during maturation. Next we will identify novel maturation-associated miRNAs through RNA-seq, and we will assess their function using over-expression, depletion and novel miRNA sensors.
Pluripotent human stem cells allow us to study events in early human development, such as formation of the cardiovascular system, and they may provide building blocks for tissue repair. This project will identify novel factors that control the differentiation of hESCs to cardiac muscle cells and regulate their subsequent maturation into working muscle.
|Hofsteen, Peter; Robitaille, Aaron Mark; Strash, Nicholas et al. (2018) ALPK2 Promotes Cardiogenesis in Zebrafish and Human Pluripotent Stem Cells. iScience 2:88-100|
|Kim, Yong Kyun; Refaeli, Ido; Brooks, Craig R et al. (2017) Gene-Edited Human Kidney Organoids Reveal Mechanisms of Disease in Podocyte Development. Stem Cells 35:2366-2378|
|Artoni, Filippo; Kreipke, Rebecca E; Palmeira, Ondina et al. (2017) Loss of foxo rescues stem cell aging in Drosophila germ line. Elife 6:|
|Palpant, Nathan J; Pabon, Lil; Friedman, Clayton E et al. (2017) Generating high-purity cardiac and endothelial derivatives from patterned mesoderm using human pluripotent stem cells. Nat Protoc 12:15-31|
|Yang, Xiulan; Murry, Charles E (2017) One Stride Forward: Maturation and Scalable Production of Engineered Human Myocardium. Circulation 135:1848-1850|
|Kadota, Shin; Pabon, Lil; Reinecke, Hans et al. (2017) In Vivo Maturation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Neonatal and Adult Rat Hearts. Stem Cell Reports 8:278-289|
|Nakamura, Paul A; Shimchuk, Andy A; Tang, Shibing et al. (2017) Small molecule Photoregulin3 prevents retinal degeneration in the RhoP23H mouse model of retinitis pigmentosa. Elife 6:|
|Moody, James D; Levy, Shiri; Mathieu, Julie et al. (2017) First critical repressive H3K27me3 marks in embryonic stem cells identified using designed protein inhibitor. Proc Natl Acad Sci U S A 114:10125-10130|
|Mathieu, Julie; Ruohola-Baker, Hannele (2017) Metabolic remodeling during the loss and acquisition of pluripotency. Development 144:541-551|
|Rabinowitz, Jeremy S; Robitaille, Aaron M; Wang, Yuliang et al. (2017) Transcriptomic, proteomic, and metabolomic landscape of positional memory in the caudal fin of zebrafish. Proc Natl Acad Sci U S A 114:E717-E726|
Showing the most recent 10 out of 114 publications