A major focus of research for the next years will be to understand on a more detailed level the molecular mechanisms driving the reprogramming of somatic cells to pluripotent IPS cells. 1. Single-cell analysis of gene expression during cellular reprogramming An unresolved issue is whether activation of specific genes can predict early in the reprogramming process whether a given cell will develop into an iPS cell. Single RNA molecule detection methods will be Used to: a. assess whether a hierarchical program of gene expression leads to IPS cell formation or whether the process is entirely stochastic as suggested by previous observations. b. define markers that at early stages of reprogramming allow the prospective identification of cells that will generate IPS cells. For this, GFP will be inserted into candidate genes to give a marker for prospective isolation of IPS precursors. 2. Stoichiometry of reprogramming factors and quality of iPS cells mRNA mediated reprogramming will be used to systematically titrate the various factors for investigating the effect of factor stoichiometry on the biological properties of the IPS cells. This will allow the optimization of reprogramming with the goal of generating high quality genetically unmodified iPS cells. 3. Transdifferentiation of somatic cells to cells of different lineages Different somatic donor cells such as liver cells and skin keratinocytes will be used for direct conversion into neural precursors and neurons. Stringent reporters will allow the retrospective confirmation of the endodermal arid ectodermal donor cell type.

Public Health Relevance

The disease in the dish approach, based on the IPS technology, is attractive for studying human diseases and for developing novel therapies. However, epigenetic and biological differences between individual IPS cells pose potentially serious hurdles for implementing this approach for research and therapy. This program uses stringent criteria to define the parameters that assure the generation of high quality iPS cells.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37HD045022-11
Application #
8528901
Study Section
Special Emphasis Panel (NSS)
Program Officer
Ravindranath, Neelakanta
Project Start
2003-07-28
Project End
2018-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
11
Fiscal Year
2013
Total Cost
$845,056
Indirect Cost
$411,694
Name
Whitehead Institute for Biomedical Research
Department
Type
DUNS #
120989983
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Wiehle, Laura; Raddatz, Günter; Musch, Tanja et al. (2016) Tet1 and Tet2 Protect DNA Methylation Canyons against Hypermethylation. Mol Cell Biol 36:452-61
Stelzer, Yonatan; Wu, Hao; Song, Yuelin et al. (2016) Parent-of-Origin DNA Methylation Dynamics during Mouse Development. Cell Rep 16:3167-80
Liu, X Shawn; Wu, Hao; Ji, Xiong et al. (2016) Editing DNA Methylation in the Mammalian Genome. Cell 167:233-247.e17
Banerjee, Abhishek; Rikhye, Rajeev V; Breton-Provencher, Vincent et al. (2016) Jointly reduced inhibition and excitation underlies circuit-wide changes in cortical processing in Rett syndrome. Proc Natl Acad Sci U S A 113:E7287-E7296
Muffat, Julien; Li, Yun; Yuan, Bingbing et al. (2016) Efficient derivation of microglia-like cells from human pluripotent stem cells. Nat Med 22:1358-1367
Ji, Xiong; Dadon, Daniel B; Powell, Benjamin E et al. (2016) 3D Chromosome Regulatory Landscape of Human Pluripotent Cells. Cell Stem Cell 18:262-75
Stelzer, Yonatan; Jaenisch, Rudolf (2015) Monitoring Dynamics of DNA Methylation at Single-Cell Resolution during Development and Disease. Cold Spring Harb Symp Quant Biol :
Cimmino, Luisa; Dawlaty, Meelad M; Ndiaye-Lobry, Delphine et al. (2015) TET1 is a tumor suppressor of hematopoietic malignancy. Nat Immunol 16:653-62
Zhao, Zhigang; Chen, Li; Dawlaty, Meelad M et al. (2015) Combined Loss of Tet1 and Tet2 Promotes B Cell, but Not Myeloid Malignancies, in Mice. Cell Rep 13:1692-704
Stelzer, Yonatan; Shivalila, Chikdu Shakti; Soldner, Frank et al. (2015) Tracing dynamic changes of DNA methylation at single-cell resolution. Cell 163:218-29

Showing the most recent 10 out of 67 publications