title: Reprogramming of enteroendocrine K cells to pancreatic β-cells through the combined expression of Nkx6.1 and Neurogenin3, and reaggregation in suspension culture.
abstract: Recent studies have demonstrated that adult cells such as pancreatic exocrine cells can be converted to pancreatic β-cells in a process called cell reprogramming. Enteroendocrine cells and β-cells share similar pathways of differentiation during embryonic development. Notably, enteroendocrine K cells express many of the key proteins found in β-cells. Thus, K cells could be reprogrammed to β-cells under certain conditions. However, there is no clear evidence on whether these cells convert to β-cells. K cells were selected from STC-1 cells, an enteroendocrine cell line expressing multiple hormones. K cells were found to express many genes of transcription factors crucial for islet development and differentiation except for Nkx6.1 and Neurogenin3. A K cell clone stably expressing Nkx6.1 (Nkx6.1(+)-K cells) was established. Induction of Neurogenin3 expression in Nkx6.1(+)-K cells, by either treatment with a γ-secretase inhibitor or infection with a recombinant adenovirus expressing Neurogenin3, led to a significant increase in Insulin1 mRNA expression. After infection with the adenovirus expressing Neurogenin3 and reaggregation in suspension culture, about 50% of Nkx6.1(+)-K cells expressed insulin as determined by immunostaining. The intracellular insulin content was increased markedly. Electron microscopy revealed the presence of insulin granules. However, glucose-stimulated insulin secretion was defective, and there was no glucose lowering effect after transplantation of these cells in diabetic mice. In conclusion, we demonstrated that K cells could be reprogrammed partially to β-cells through the combined expression of Nkx6.1 and Neurogenin3, and reaggregation in suspension culture.
authors.affiliation: Division of Endocrinology & Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Esder
authors.initials: E
authors.lastname: Lee
authors.affiliation: Division of Endocrinology & Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Gyeong Ryul
authors.initials: GR
authors.lastname: Ryu
authors.affiliation: Division of Endocrinology & Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Sung-Dae
authors.initials: SD
authors.lastname: Moon
authors.affiliation: Division of Endocrinology & Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Seung-Hyun
authors.initials: SH
authors.lastname: Ko
authors.affiliation: Division of Endocrinology & Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Yu-Bae
authors.initials: YB
authors.lastname: Ahn
authors.affiliation: Division of Endocrinology & Metabolism, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea. Electronic address: kihos@catholic.ac.kr.
authors.forename: Ki-Ho
authors.initials: KH
authors.lastname: Song
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000379:methods*; D065150:Cellular Reprogramming; D019858:Enteroendocrine Cells / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008819:Mice, Nude; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D051381:Rats; D013535:Suspensions
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D013535:Suspensions; C407737:Neurog3 protein, mouse; C491327:Nkx6-1 protein, rat
keywords: 4′,6′-diamidino-2-phenylindole; Adv; DAPI; DAPT; DMEM; Differentiation; Dulbecco's Modified Eagle's Medium; Enteroendocrine cells; GFP; GIP; GIPP; GLP-1; GLUT2; ITS; Islet; Islet transplantation; K cells; KRB; Krebs Ringer Buffer; MOI; N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester; NGN3; Neurogenin 3; PC; PDX1; Reprogramming; adenovirus; glucagon-like peptide-1; glucose transporter 2; glucose-dependent insulinotropic polypeptide; glucose-dependent insulinotropic polypeptide promoter; green fluorescent protein; insulin/transferring/sodium selenite; multiplicity of infection; pancreatic and duodenal homeobox-1; prohormone convertase; β-Cells