This application seeks to investigate the mechanisms underlying cellular reprogramming, i.e. the conversion of adult cells into pluripotent stem cells. We will utilize transcription factor-mediated reprogramming into induced pluripotent stem cells (iPSCs) as a tool to pursue three major Aims.
In Aim 1, we will (i) test whether iPSCs derived from fibroblasts, hematopoietic and myogenic cells are transcriptionally, epigenetically and functionally distinct, (ii) study the mechanism for the increased reprogramming efficiency of progenitors, and (iii) determine if iPSC formation introduces genetic mutations into cells. Addressing the epigenetic and genetic integrity of iPSCs will be crucial for any potential therapeutic applications of this technology and may identify the most suitable cell type for generating patient-specific iPSCs.
In Aim 2, we will (i) map the transcriptional and epigenetic changes in intermediate cell populations undergoing reprogramming, (ii) test if reprogramming differentiated cells into pluripotent cells recapitulates stages of normal development, and (iii) perform a gain and loss-of-function screen, respectively, to identify novel regulators of reprogramming.
This aim will identify new molecules important during reprogramming, whose manipulation may facilitate the efficient and safe generation of patient-specific iPSCs.
In Aim 3, we will investigate the functionality of iPSCs compared with ESCs using in vitro and in vivo assays. Specifically, we will (i) assess whether neural stem cells and fibroblasts derived from iPSCs and ESCs show similar growth and differentiation characteristics in vitro, (ii) test if iPSC-derived hematopoietic stem cells are as potent as ESC-derived hematopoietic stem cells upon serial bone marrow transplantation, and (iii) produce and age entirely ESC and iPSC-derived mice to test if iPSC-derived animals age prematurely or develop cancer.
This aim will assess the safety and long-term consequences of iPSCs-derived mature cells in vivo, a prerequisite for using iPSC technology in human cell therapy.
The goal of our lab is to dissect the mechanisms of cellular reprogramming by using induced pluripotent stem cells (iPSCs) as a tool. In this proposal, we will (i) study the role of the somatic cell-of-origin during cellular reprogramming into iPSCs, (ii) map the transcriptional and epigenetic events that occur in intermediate cells undergoing reprogramming, and (iii) compare the developmental and differentiation potentials of iPSCs and embryonic stem cells. Collectively, these experiments will give insight into the mechanisms of transcription-factor-mediated reprogramming and provide crucial information on the efficacy and safety of iPSC production for studying and potentially treating diseases.
|Miyoshi, Norikatsu; Stel, Jente M; Shioda, Keiko et al. (2016) Erasure of DNA methylation, genomic imprints, and epimutations in a primordial germ-cell model derived from mouse pluripotent stem cells. Proc Natl Acad Sci U S A 113:9545-50|
|Borkent, Marti; Bennett, Brian D; Lackford, Brad et al. (2016) A Serial shRNA Screen for Roadblocks to Reprogramming Identifies the Protein Modifier SUMO2. Stem Cell Reports 6:704-16|
|Cheloufi, Sihem; Elling, Ulrich; Hopfgartner, Barbara et al. (2015) The histone chaperone CAF-1 safeguards somatic cell identity. Nature 528:218-24|
|Etchegaray, Jean-Pierre; Chavez, Lukas; Huang, Yun et al. (2015) The histone deacetylase SIRT6 controls embryonic stem cell fate via TET-mediated production of 5-hydroxymethylcytosine. Nat Cell Biol 17:545-57|
|Bar-Nur, Ori; Verheul, Cassandra; Sommer, Andreia G et al. (2015) Lineage conversion induced by pluripotency factors involves transient passage through an iPSC stage. Nat Biotechnol 33:761-8|
|West, Jason A; Cook, April; Alver, Burak H et al. (2014) Nucleosomal occupancy changes locally over key regulatory regions during cell differentiation and reprogramming. Nat Commun 5:4719|
|Bar-Nur, Ori; Brumbaugh, Justin; Verheul, Cassandra et al. (2014) Small molecules facilitate rapid and synchronous iPSC generation. Nat Methods 11:1170-6|
|Schwarz, Benjamin A; Bar-Nur, Ori; Silva, JosÃ© C R et al. (2014) Nanog is dispensable for the generation of induced pluripotent stem cells. Curr Biol 24:347-50|
|Wang, Li; Miao, Yi-Liang; Zheng, Xiaofeng et al. (2013) The THO complex regulates pluripotency gene mRNA export and controls embryonic stem cell self-renewal and somatic cell reprogramming. Cell Stem Cell 13:676-90|
|Brack, Andrew S; Hochedlinger, Konrad (2013) ISSCR 2013: back to Bean Town. Stem Cell Reports 1:479-85|
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