Previous groundbreaking discoveries have made it possible to grow pluripotent embryonic stem (ES) cells in vitro and to reprogram adult cells back to a pluripotent state. These advances have opened new opportunities for regenerative medicine by providing the cellular starting point from which we can derive adult cell types with physiological properties to study and treat human disease. Attaining cell types suitable for direct therapeutic application or as cellular models of human disease for drug development both require a substantial level of similarly to the in vivo cellular counterpart. Most often, investigaors attempt to attain the best in vivo mimics by replicating key aspects of a cell's developmental progression through embryogenesis. Given the appreciation that epigenetic effects are cumulative and progressive through embryogenesis, a central hypothesis of this proposal is that the initial steps in lineage specification are critical, and epigenetic mistakes made during exit from pluripotency can have undesirable effects even after cells have reached the targeted differentiated state. To address this problem, we aim to elucidate the mechanisms that eliminate or decommission ES cell- specific gene regulatory elements (i.e. enhancers) from lineage specified cell types. We discovered that a sequence-specific DNA-binding transcription factor, Tcf7l1, binds specifically to enhancers in ES cells that are targeted for decommissioning. Given our previous findings that showed Tcf7l1 stimulated differentiation of ES cells, we propose testing the hypothesis that Tcf7l1 is necessary to decommission ES specific enhancers. The proposal will further determine how this effect of Tcf7l1 changes the responses of cells to extracellular signals, and measure how important it is for genuine lineage specification using established in vitro differentiation protocols. The immediate outcome of completing the proposed research will be a better understanding of how cells exit the pluripotent state, and a demonstration of how defects during early steps can diminish the efficiency of subsequent stages of differentiation. Finally, given the oncogenic potential of accessible ES- specific enhancers in adult cells, the importance of the proposed work for increasing the safety of cell-based therapies should be appreciated.

Public Health Relevance

The proposed research will elucidate the earliest steps in the progression of embryonic stem cells to a differentiated, adult cell type. This knowledge will be important for understanding how an identical genome can used very differently in distinct cell types and provide insight enabling more efficient and safer manipulation of stem cells for regenerative medicine.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD081534-05
Application #
9501742
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Mukhopadhyay, Mahua
Project Start
2014-08-15
Project End
2019-05-31
Budget Start
2018-06-01
Budget End
2019-05-31
Support Year
5
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Biochemistry
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Clarke, Ryan; Heler, Robert; MacDougall, Matthew S et al. (2018) Enhanced Bacterial Immunity and Mammalian Genome Editing via RNA-Polymerase-Mediated Dislodging of Cas9 from Double-Strand DNA Breaks. Mol Cell 71:42-55.e8
Cha, Boksik; Geng, Xin; Mahamud, Md Riaj et al. (2018) Complementary Wnt Sources Regulate Lymphatic Vascular Development via PROX1-Dependent Wnt/?-Catenin Signaling. Cell Rep 25:571-584.e5
Sierra, Robert A; Hoverter, Nathan P; Ramirez, Ricardo N et al. (2018) TCF7L1 suppresses primitive streak gene expression to support human embryonic stem cell pluripotency. Development 145:
Huang, Shuo; Fantini, Damiano; Merrill, Bradley J et al. (2017) DDB2 Is a Novel Regulator of Wnt Signaling in Colon Cancer. Cancer Res 77:6562-6575
Shy, Brian R; MacDougall, Matthew S; Clarke, Ryan et al. (2016) Co-incident insertion enables high efficiency genome engineering in mouse embryonic stem cells. Nucleic Acids Res 44:7997-8010
Gaston-Massuet, Carles; McCabe, Mark J; Scagliotti, Valeria et al. (2016) Transcription factor 7-like 1 is involved in hypothalamo-pituitary axis development in mice and humans. Proc Natl Acad Sci U S A 113:E548-57