Embryonic stem (ES) cells not only hold considerable promise for the treatment of a number of devastating diseases (e.g. cardiovascular diseases, neurodegenerative diseases, diabetes and cancers), they also provide an ideal tractable culture system for studying early embryonic development of all cell types and modeling human diseases. Harnessing these potentials of ESCs will require an improved ability to manipulate their self-renewal and differentiation, and a better understanding of the signaling pathways that control their fate.
To address the critical challenges in hESC culture (i.e. robust self-renewal and single cell survival in chemically defined conditions), we propose to (1) optimize two previously identified and functionally characterized novel small molecules (from high throughput screens in hESCs) via medicinal chemistry for much improved self-renewal and survival of hESCs, and screen additional 85,000 diverse compounds to identify new small molecules with different mechanisms of action that can maintain self-renewal of hESCs in the absence of growth factors, or promote single cell survival of hESCs under the chemically defined media. We will further confirm and characterize their effects and activities via various in-depth cellular/biochemical assays, and carry out structure-activity-relationship (SAR) studies of the selected hit compounds (that especially act on different targets/mechanisms as compared to the two molecules in Aim 1) to optimize their potency and specificity. Collectively, the studies described in this proposal will provide novel chemical tools for better understanding and controlling hESC, and may ultimately allow development of therapeutics employing hESCs for treating diseases.
The proposed studies will provide new chemical tools and technologies and substantial knowledge for better understanding and controlling hESC self-renewal and differentiation (e.g. development of robust self-renewal and clonal expansion conditions for hESCs) , and may ultimately allow development of therapeutics employing hESCs for treating diseases.