To exploit the full potential of human pluripotent stem (hPS) cells for regenerative medicine, developmental biology, and drug discovery, defined conditions for hPS cell propagation and differentiation must be reproducible and therefore chemically defined. To this end, the chemical cues that instruct hPS cells to make specific decisions must be determined. While strategies to identify the soluble cues are well established, an understanding of how insoluble signals guide cell fate decisions is lacking. Chemical synthesis provides access to well-defined surfaces and materials that vary in binding epitope identity, density, and in physicochemical properties. The three Specific Aims have been devised to leverage synthetic methods from chemistry and chemical biology to generate tailored materials and surfaces that promote hPS cell expansion or differentiation to desired cell types. In the past grant period, chemically defined peptide- substituted surfaces that promote hPS cell propagation were identified. The most effective substrates interact with hPS cell surface glycosaminoglycans. The experiments proposed in Aim 1 were conceived to test the hypothesis that engagement of hPS glycosaminoglycans contributes to pluripotency.
In Aim 2, we shall employ our surface array screen to identify insoluble signals (peptide- or small molecule-derived) that act in concert with soluble signals to direct hPS cell differentiation.
In Aim 3, we shall synthesize a series of biocompatible polymers that vary in cell binding groups, pore size, and elasticity to address how physicochemical cues influence cell fate decisions. A novel feature of the approach is the use of an array strategy to identify new surfaces that act on hPS cells to direct their proliferation and differentiation. The proposed experiments leverage this surface array screen not only to identify functional surfaces but also to devise and test novel hypotheses regarding the signaling pathways and molecular mechanisms that underlie hPS self-renewal and differentiation. Progress in understanding how hPS cells respond to distinct combinations of soluble, insoluble, and mechanical cues will be transformative: It will advance applications of hPS cells to human disease.

Public Health Relevance

Human pluripotent stem cells can be used to identify drugs for key human diseases, and they are promising potential sources of more specialized human cells (e.g., heart cells or brain cells) for treating disease. To obtain each desired type of specialized cell, the right combination of chemical cues must be given, such that hPS cells reproducibly differentiate into the specific cell type needed. The research proposed is designed to provide fundamental knowledge regarding how combinations of chemical cues influence hPS cell propagation and differentiation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM049975-19
Application #
8644264
Study Section
Special Emphasis Panel (ZRG1-SBCA-U (02))
Program Officer
Marino, Pamela
Project Start
1993-07-01
Project End
2017-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
19
Fiscal Year
2014
Total Cost
$422,381
Indirect Cost
$104,639
Name
University of Wisconsin Madison
Department
Biochemistry
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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Fishman, Joshua M; Kiessling, Laura L (2013) Synthesis of functionalizable and degradable polymers by ring-opening metathesis polymerization. Angew Chem Int Ed Engl 52:5061-4
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Garber, Kathleen C A; Wangkanont, Kittikhun; Carlson, Erin E et al. (2010) A general glycomimetic strategy yields non-carbohydrate inhibitors of DC-SIGN. Chem Commun (Camb) 46:6747-9
Kiessling, Laura L; Splain, Rebecca A (2010) Chemical approaches to glycobiology. Annu Rev Biochem 79:619-53
Klim, Joseph R; Li, Lingyin; Wrighton, Paul J et al. (2010) A defined glycosaminoglycan-binding substratum for human pluripotent stem cells. Nat Methods 7:989-94
Kolonko, Erin M; Pontrello, Jason K; Mangold, Shane L et al. (2009) General synthetic route to cell-permeable block copolymers via ROMP. J Am Chem Soc 131:7327-33
Allen, Matthew J; Wangkanont, Kittikhun; Raines, Ronald T et al. (2009) ROMP from ROMP: A New Approach to Graft Copolymer Synthesis. Macromolecules 42:4023-4027

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