Grant Award Details
Grant Application Details
- Chemical Genetic Approach to Production of hESC-derived Cardiomyocytes
Adult heart muscle cells retain negligible proliferative capacity and this underlies the inability of the heart to replace muscle cells that are lost to injury, such as infarct, and underlies progression to heart failure. To date, no stem cell therapiy has produced significant cardiomyocyte replacement. Instead, transplanted cells, if they persist at all, produce endothelial cells or fibroblasts and the ameliorating effects on heart function that have been reported have been achieved by improving contractility, perfusion or other processes that are impaired in the failing heart. This proposal is to develop specific reagents and ultimately drugs to stimulate regeneration. Our approach integrates advances in stem cell biology, high-throughput (HT) biology, informatics and proteomics to identify small molecules, proteins and signal transduction pathways that control heart muscle formation from human embryonic stem cells (hESCs). High throughput assays will be developed and implemented to identify genes, signaling proteins, and small molecules that that control important steps in the differentiation, proliferation, and maturation of cardiac cells. Computer modeling and informatics will be used to identify and validate the signaling pathways that direct these critical processes. The discovery of small molecules and pathways will lead to protocols for 1) efficient directed differentiation of cardiomyogenic precursors from hESCs for research into transplantation and toxicology, 2) biotech reagents to stimulate cardiomyocyte renewal through directed differentiation of hESCs, and 3) cellular targets or lead compounds to develop drugs that stimulate regeneration from endogenous cells.
This proposal is a multidisciplinary collaboration among stem cell biologists, chemists, and engineers to address a critical problem that limits the widespread use of hESC for cardiology applications. Developing the multidisciplinary technology and overcoming the hurdles to application of hESCs to biotech and clinic will benefit California in many ways, including:
Research to discover novel tools to stimulate heart muscle regeneration from hESCs is clinically important. Cardiovascular disease is the single largest cause of death in the U.S. and the assays we will develop and the reagents themselves will be useful tools to direct cardiomyocyte regeneration. This will speed the translation of hESCs to the clinic, specifically by stimulating production of cardiomyocytes and potentially by enhancing their integration and function after engraftment.
Heart regeneration from hESCs probably uses similar cellular proteins and signaling pathways as regeneration of cardiomyocytes from other sources, thus, this research might be broadly applicable to heart muscle repair. Regeneration from endogenous cells remains controversial but these tools should be useful reagents to study and hopefully stimulate endogenous repair.
Bringing the diverse people together (chemists, cell biologists, and engineers) to address a stem cell problem forges new links in the academic community that should be capable of opening new areas of research. These new areas of research will be a important legacy of the stem cell initiative and promises to invigorate academic research.
The technology that we are developing applies the new discipline of chemical biology to stem cell biology, and the merger promises to spin off new areas of investigation and biotech products with the potential to benefit the practice of medicine and the local economy.
Lastly, supporting the leading edge technology and the collaboration will build the California infrastructure of high throughput chemical library screening so that it can be focused on other areas of biomedical research, both stem cell and non-stem cell.
- Nature (2014) Inhibition of miR-25 improves cardiac contractility in the failing heart. (PubMed: 24670661)
- J Med Chem (2012) Wnt Inhibition Correlates with Human Embryonic Stem Cell Cardiomyogenesis: A Structure-Activity Relationship Study Based on Inhibitors for the Wnt Response. (PubMed: 22191557)
- Cell Stem Cell (2012) Small Molecule-Mediated TGF-beta Type II Receptor Degradation Promotes Cardiomyogenesis in Embryonic Stem Cells. (PubMed: 22862949)
- J Med Chem (2012) Synthesis and SAR of b-Annulated 1,4-Dihydropyridines Define Cardiomyogenic Compounds as Novel Inhibitors of TGFbeta Signaling. (PubMed: 23130626)
- Genes Dev (2012) Whole-genome microRNA screening identifies let-7 and mir-18 as regulators of germ layer formation during early embryogenesis. (PubMed: 23152446)
- Curr Protoc Stem Cell Biol (2012) Serum-free generation of multipotent mesoderm (kdr(+)) progenitor cells in mouse embryonic stem cells for functional genomics screening. (PubMed: 23154934)
- Circ Res (2011) Small-Molecule Inhibitors of the Wnt Pathway Potently Promote Cardiomyocytes From Human Embryonic Stem Cell-Derived Mesoderm. (PubMed: 21737789)
- Mol Cell (2011) Fine-tuning of Drp1/Fis1 availability by AKAP121/Siah2 regulates mitochondrial adaptation to hypoxia. (PubMed: 22099302)
- J Cardiovasc Transl Res (2011) A chemical biology approach to myocardial regeneration. (PubMed: 21424858)
- Stem Cells Dev (2010) Non-cardiomyocytes influence the electrophysiological maturation of human embryonic stem cell-derived cardiomyocytes during differentiation. (PubMed: 20001453)
- Ann N Y Acad Sci (2010) Cardiac myocyte force development during differentiation and maturation. (PubMed: 20201894)
- Proc Natl Acad Sci U S A (2010) Alternative splicing regulates mouse embryonic stem cell pluripotency and differentiation. (PubMed: 20498046)
- PLoS Comput Biol (2009) Alternative splicing in the differentiation of human embryonic stem cells into cardiac precursors. (PubMed: 19893621)
- Circulation (2009) Electrophysiological challenges of cell-based myocardial repair. (PubMed: 20008740)
- PLoS One (2009) Lentiviral vectors and protocols for creation of stable hESC lines for fluorescent tracking and drug resistance selection of cardiomyocytes. (PubMed: 19352491)
- Pediatr Cardiol (2009) Natural and synthetic regulators of embryonic stem cell cardiogenesis. (PubMed: 19319460)
- PLoS One (2008) Contrasting expression of keratins in mouse and human embryonic stem cells. (PubMed: 18941637)
- Dev Biol (2008) A novel activity of the Dickkopf-1 amino terminal domain promotes axial and heart development independently of canonical Wnt inhibition. (PubMed: 18840423)
- Assay Drug Dev Technol (2008) A comparative analysis of standard microtiter plate reading versus imaging in cellular assays. (PubMed: 18795873)