title: High-throughput Functional Genomics Identifies Regulators of Primary Human Beta Cell Proliferation.
abstract: The expansion of cells for regenerative therapy will require the genetic dissection of complex regulatory mechanisms governing the proliferation of non-transformed human cells. Here, we report the development of a high-throughput RNAi screening strategy specifically for use in primary cells and demonstrate that silencing the cell cycle-dependent kinase inhibitors CDKN2C/p18 or CDKN1A/p21 facilitates cell cycle entry of quiescent adult human pancreatic beta cells. This work identifies p18 and p21 as novel targets for promoting proliferation of human beta cells and demonstrates the promise of functional genetic screens for dissecting therapeutically relevant state changes in primary human cells.
authors.affiliation: From the Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario K1H 8L1.
authors.forename: Karine
authors.initials: K
authors.lastname: Robitaille
authors.affiliation: the Sunnybrook Research Institute, Toronto, Ontario M4N 3M5.
authors.forename: Jillian L
authors.initials: JL
authors.lastname: Rourke
authors.affiliation: From the Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario K1H 8L1.
authors.forename: Joanne E
authors.initials: JE
authors.lastname: McBane
authors.affiliation: From the Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario K1H 8L1, the Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario K1H 8M5.
authors.forename: Accalia
authors.initials: A
authors.lastname: Fu
authors.affiliation: From the Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario K1H 8L1.
authors.forename: Stephen
authors.initials: S
authors.lastname: Baird
authors.affiliation: the Sunnybrook Research Institute, Toronto, Ontario M4N 3M5.
authors.forename: Qiujiang
authors.initials: Q
authors.lastname: Du
authors.affiliation: the Clinical Islet Transplant Program, University of Alberta, 8215-112 Street, Edmonton, Alberta T6G 2C8, and.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Kin
authors.affiliation: the Clinical Islet Transplant Program, University of Alberta, 8215-112 Street, Edmonton, Alberta T6G 2C8, and.
authors.forename: A M James
authors.initials: AM
authors.lastname: Shapiro
authors.affiliation: the Sunnybrook Research Institute, Toronto, Ontario M4N 3M5, the Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada robert.screaton@sri.utoronto.ca.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Screaton
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000416:Alberta; D015415:Biomarkers / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D050763:Cyclin-Dependent Kinase Inhibitor p18 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D005240:Feasibility Studies; D005260:Female; D023281:Genomics / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010865:Pilot Projects; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014019:Tissue Donors; D055815:Young Adult
chemical_list: D015415:Biomarkers; C493956:CDKN1A protein, human; C493971:CDKN2C protein, human; D050763:Cyclin-Dependent Kinase Inhibitor p18; D050759:Cyclin-Dependent Kinase Inhibitor p21; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: RNA interference (RNAi); beta cell (β-cell); cell cycle; high throughput screening (HTS); pancreatic islet; primary human cells