Delineating key factors that regulate early pancreas development is crucial to our long-term pursuit of understanding disease mechanisms and developing human pluripotent stem cell (hPSC) based ?-cell replacement therapies for diabetes. However, a complete landscape of signaling cues and transcription factors required for pancreas specification remains unclear. Using hPSC directed differentiation and CRISPR/Cas-mediated genome editing, recently published work from the Huangfu lab has identified critical new requirements for RFX6, GATA6 and GATA4 (genes known to be associated with neonatal and adult-onset diabetes) during human pancreatic differentiation. In unpublished work, we have further uncovered a novel, dose-dependent requirement for FOXA2, a gene associated with diabetes and hyperinsulinism, in pancreatic differentiation. Here in this collaboration between the Huangfu, Leslie and Pe?er labs, we will undertake complementary genetic, genomic and computational approaches and utilize hPSC differentiation to dissect human pancreatic development. We will utilize genetic approach to create precise hPSC disease models and interrogate complex genetic interactions underlying disease phenotypes, and employ genomic approaches including ChIP-seq and ATAC-seq analyses and cutting-edge single-cell transcriptomics to understand regulators of human pancreatic development and ? cell function. Our findings will enhance the understanding of human pancreatic development and disease, and facilitate the development of improved hPSC directed differentiation protocols for the generation of functional ? cells for disease study and treatment.
This project aims to combine sophisticated genetic manipulation, state-of-art single-cell technology, and human pluripotent stem cell (hPSC) directed differentiation to uncover mechanisms that control pancreatic ? cell differentiation and function. Our human-cell based system will provide unique mechanistic understanding of neonatal and adult-onset diabetes not evident in mouse models, and facilitate the development of improved hPSC directed differentiation protocols for the generation of functional insulin-secreting ? cells for diabetes cell-replacement therapy.
|Verma, Nipun; Pan, Heng; Doré, Louis C et al. (2018) TET proteins safeguard bivalent promoters from de novo methylation in human embryonic stem cells. Nat Genet 50:83-95|
|Teijeiro, Virginia; Yang, Dapeng; Majumdar, Sonali et al. (2018) DICER1 Is Essential for Self-Renewal of Human Embryonic Stem Cells. Stem Cell Reports 11:616-625|
|Shukla, Abhijit; Huangfu, Danwei (2018) Decoding the noncoding genome via large-scale CRISPR screens. Curr Opin Genet Dev 52:70-76|
|Amin, Sadaf; Cook, Brandoch; Zhou, Ting et al. (2018) Discovery of a drug candidate for GLIS3-associated diabetes. Nat Commun 9:2681|
|Pulecio, Julian; Verma, Nipun; Mejía-Ramírez, Eva et al. (2017) CRISPR/Cas9-Based Engineering of the Epigenome. Cell Stem Cell 21:431-447|
|Verma, Nipun; Zhu, Zengrong; Huangfu, Danwei (2017) CRISPR/Cas-Mediated Knockin in Human Pluripotent Stem Cells. Methods Mol Biol 1513:119-140|
|Soh, Chew-Li; Huangfu, Danwei (2017) CRISPR/Cas9-Mediated Mutagenesis of Human Pluripotent Stem Cells in Defined Xeno-Free E8 Medium. Methods Mol Biol 1498:57-78|
|Shi, Zhong-Dong; Lee, Kihyun; Yang, Dapeng et al. (2017) Genome Editing in hPSCs Reveals GATA6 Haploinsufficiency and a Genetic Interaction with GATA4 in Human Pancreatic Development. Cell Stem Cell 20:675-688.e6|
|Wang, Qiong; Zou, Yilong; Nowotschin, Sonja et al. (2017) The p53 Family Coordinates Wnt and Nodal Inputs in Mesendodermal Differentiation of Embryonic Stem Cells. Cell Stem Cell 20:70-86|
|Shi, Zhong-Dong; Soh, Chew-Li; Zhu, Zengrong et al. (2017) Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development. J Vis Exp :|
Showing the most recent 10 out of 17 publications