Understanding how human embryonic stem (ES) cells can proliferate without limit and yet retain the ability to differentiate to any cell type is the theme that links the three individual projects of this proposal. Project 1 will identify novel histone modifications in human ES cells using a new mass spectrometry technique that allows an unprecedented ability to identify and map posttranslational protein modifications. Histone modifications in human ES cells will be identified globally, at promoters, and at select genes directly regulated by critical pluripotency factors. We will also examine the role of histone H3 variants in establishing long-term epigenetic memory. These studies will determine whether there is a novel histone code in pluripotent cells and determine how histone modifications change dynamically during differentiation. Project 2 examines the critical events that occur in the window of time during which human ES cells commit eprogram differentiated cells to a pluripotent state. We have previously reported that when myeloid cells are fused with human ES cells, the myeloid nucleus is reprogrammed to an ES cell state, indicating that transacting factors in ES cells are sufficient to mediate nuclear reprogramming. Our preliminary results suggest that over expressing combinations of human ES cell-enriched genes can reprogram myeloid cells, and this project will optimize this reprogramming. The combination of these projects will provide an increased understanding of the pluripotent state and the basic processes by which a cell can leave or return to that state. Such an understanding will be important to transplantation and regenerative medicine. Lay Description: Human ES cells are special because they can grow without limit and can give rise to all other cell types. Here we will try to understand why human ES cells have this remarkable developmental potential, and develop conditions to convert a cell with a more limited potential to an ES cell. Such reprogramming has implications for transplantation and regenerative medicine. ????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????

Public Health Relevance

Lay Description. Human ES cells are special because they can grow without limit and can give rise to all other cell types. Here we will try to understand why human ES cells have this remarkable developmental potential, and develop conditions to convert a cell with a more limited potential to an ES cell. Such reprogramming has implications for transplantation and regenerative medicine.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM081629-05
Application #
8338841
Study Section
Special Emphasis Panel (ZGM1-GDB-8 (SC))
Program Officer
Haynes, Susan R
Project Start
2008-08-01
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2014-07-31
Support Year
5
Fiscal Year
2012
Total Cost
$1,661,048
Indirect Cost
$295,586
Name
Morgridge Institute for Research, Inc.
Department
Type
DUNS #
012420082
City
Madison
State
WI
Country
United States
Zip Code
53715
Sheynkman, Gloria M; Johnson, James E; Jagtap, Pratik D et al. (2014) Using Galaxy-P to leverage RNA-Seq for the discovery of novel protein variations. BMC Genomics 15:703
Bailey, Derek J; McDevitt, Molly T; Westphall, Michael S et al. (2014) Intelligent data acquisition blends targeted and discovery methods. J Proteome Res 13:2152-61
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Brumbaugh, Justin; Russell, Jason D; Yu, Pengzhi et al. (2014) NANOG is multiply phosphorylated and directly modified by ERK2 and CDK1 in vitro. Stem Cell Reports 2:18-25
Sheynkman, Gloria M; Shortreed, Michael R; Frey, Brian L et al. (2014) Large-scale mass spectrometric detection of variant peptides resulting from nonsynonymous nucleotide differences. J Proteome Res 13:228-40
Russell, Jason D; Scalf, Mark; Book, Adam J et al. (2013) Characterization and quantification of intact 26S proteasome proteins by real-time measurement of intrinsic fluorescence prior to top-down mass spectrometry. PLoS One 8:e58157
Kim, Do-Young; Scalf, Mark; Smith, Lloyd M et al. (2013) Advanced proteomic analyses yield a deep catalog of ubiquitylation targets in Arabidopsis. Plant Cell 25:1523-40
Frey, Brian L; Ladror, Daniel T; Sondalle, Samuel B et al. (2013) Chemical derivatization of peptide carboxyl groups for highly efficient electron transfer dissociation. J Am Soc Mass Spectrom 24:1710-21
Miller, Marcus J; Scalf, Mark; Rytz, Therese C et al. (2013) Quantitative proteomics reveals factors regulating RNA biology as dynamic targets of stress-induced SUMOylation in Arabidopsis. Mol Cell Proteomics 12:449-63
Sheynkman, Gloria M; Shortreed, Michael R; Frey, Brian L et al. (2013) Discovery and mass spectrometric analysis of novel splice-junction peptides using RNA-Seq. Mol Cell Proteomics 12:2341-53

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