The goal of our Program renewal is to build upon the novel and unexpected discoveries of our last grant period to dissect, perturb, and control molecules and networks that enable or restrict the conversion of a somatic cell to pluripotency. Our last grant period revealed that, during conversion to pluripotency, resident enhancers in somatic cells that maintain cell differentiation must be decommissioned and pluripotency enhancers must be gradually activated, heterochromatic domains must be disassembled, and the post- transcriptional processes of polyadenylation and protein sumoylation must be altered. By investigating these parameters collectively in the next grant period, the Plath, Zaret, and Hochedlinger labs will determine how gene-regulatory networks integrate with cell biological transitions that are crucial for timely and efficient cellular reprogramming. Our three specific projects address the following fundamental questions: 1) How are enhancer patterns reorganized during the conversion of somatic cells to pluripotency, which genomic and epigenomic features are important, and how can heterochromatin be disassembled? 2) What comprises heterochromatin and how can its constituents be antagonized to facilitate reprogramming? 3) How do post-transcriptional regulation of mRNA polyadenylation and protein sumoylation suppress the pluripotency program? Answers to these mechanistic questions will provide insights to facilitate specific avenues to enhance cellular reprogramming to pluripotency as well as direct reprogramming from one somatic fate to another. The principles we uncover will also have direct applications for understanding cell type conversions in development, disease, and regeneration. Notably, new discoveries and scientific overlap between the three P.I.'s makes this a far more synergistic and interdependent proposal than the previous submission. The Program also takes advantage of extensive collaborations between our three groups, all leaders in the field of reprogramming, with a track record of working and publishing together. Our plan to evaluate chromatin and transcriptional features and post-transcriptional mechanisms within one Program will greatly enhance a sophisticated and cohesive view of how somatic cells can change fate to become pluripotent, identify commonalities and potential differences between human and mouse models, and allow us to determine how diverse features can be modulated coordinately to boost the efficiency and faithfulness of reprogramming to pluripotency and of one somatic fate to another. An Administrative Core will ensure efficiency and data sharing, leverages existing cores at our respective stem cell institutions, offer a pilot program to bring in additional investigators supporting our goals, and integrates a Scientific Advisory Board, and make sure that our goals are achieved collaboratively.
The information generated by the combined effort of the Plath, Zaret, and Hochedlinger labs' program will provide valuable insights into the efficient, timely and faithful reprogramming of somatic cells into pluripotent stem cells. Successful completion of our specific aims will greatly facilitate the generation of reprogrammed cells and autologous derivatives for transplantation and disease modeling. Our synergistic efforts will also provide important knowledge about other forms of cellular reprogramming and further our understanding of basic mechanisms that control cell fate transitions.
|Pasque, Vincent; Karnik, Rahul; Chronis, Constantinos et al. (2018) X Chromosome Dosage Influences DNA Methylation Dynamics during Reprogramming to Mouse iPSCs. Stem Cell Reports 10:1537-1550|
|Ohashi, Minori; Korsakova, Elena; Allen, Denise et al. (2018) Loss of MECP2 Leads to Activation of P53 and Neuronal Senescence. Stem Cell Reports 10:1453-1463|
|Kaeding, Kelsey E; Zaret, Kenneth S (2018) Microsatellite enhancers can be targeted to impair tumorigenesis. Genes Dev 32:991-992|
|Allison, Thomas F; Smith, Andrew J H; Anastassiadis, Konstantinos et al. (2018) Identification and Single-Cell Functional Characterization of an Endodermally Biased Pluripotent Substate in Human Embryonic Stem Cells. Stem Cell Reports 10:1895-1907|
|Sereti, Konstantina-Ioanna; Nguyen, Ngoc B; Kamran, Paniz et al. (2018) Analysis of cardiomyocyte clonal expansion during mouse heart development and injury. Nat Commun 9:754|
|Di Stefano, Bruno; Ueda, Mai; Sabri, Shan et al. (2018) Reduced MEK inhibition preserves genomic stability in naive human embryonic stem cells. Nat Methods 15:732-740|
|Sun, Fei; Chronis, Constantinos; Kronenberg, Michael et al. (2018) Promoter-Enhancer Communication Occurs Primarily within Insulated Neighborhoods. Mol Cell :|
|Bar-Nur, Ori; Gerli, Mattia F M; Di Stefano, Bruno et al. (2018) Direct Reprogramming of Mouse Fibroblasts into Functional Skeletal Muscle Progenitors. Stem Cell Reports 10:1505-1521|
|Xie, Yuan; Lowry, William E (2018) Manipulation of neural progenitor fate through the oxygen sensing pathway. Methods 133:44-53|
|Brumbaugh, Justin; Di Stefano, Bruno; Wang, Xiuye et al. (2018) Nudt21 Controls Cell Fate by Connecting Alternative Polyadenylation to Chromatin Signaling. Cell 172:629-631|
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