Novel insulinoma cell lines produced by iterative engineering of GLUT2, glucokinase, and human insulin expression

Diabetes
S A ClarkK Normington

Abstract

Cellular engineering studies in our group are directed at creating insulin-secreting cell lines that simulate the performance of the normal islet beta-cell. The strategy described in this article involves the stepwise stable introduction of genes relevant to beta-cell performance into the RIN 1046-38 insulinoma cell line, a process that we term "iterative engineering." RIN cells stably engineered to contain multiple copies of the human insulin gene exhibit a large increase in insulin content, such that they approach the content of human islets assayed in parallel. Analysis by high-performance liquid chromatography demonstrates that these engineered cell lines process human proinsulin to mature insulin with high efficiency. Cell lines that are further engineered to express the GLUT2 and glucokinase genes demonstrate stable expression of the three transgenes for the full lifetime of the lines produced to date (6 months to 1 year in continuous culture). Transplantation of the engineered cell lines into nude rats reveals that stably integrated genes are expressed at constant levels in the in vivo environment over the full duration of experiments performed (48 days). Several endogenous genes expressed in normal beta-cells, including...Continue Reading

Citations

Nov 5, 1997·Diabetologia·S FerberC Colton
Oct 30, 1998·Molecular Medicine Today·A W Varley, R S Munford
Mar 15, 2002·Drug Discovery Today·Paul A Johnston, Patricia A Johnston
May 11, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jonathan C SchislerChristopher B Newgard
Oct 28, 2003·Human Gene Therapy·Darin E OlsonPeter M Thulé
May 26, 1999·Human Gene Therapy·L GrosF Bosch
Feb 4, 1998·Human Gene Therapy·L GrosF Bosch
Jun 7, 2007·Tissue Engineering·Lye T Lock, Emmanuel S Tzanakakis
May 20, 2006·Endocrine Journal·Aiko IshiiKennichi Kakudo
May 26, 2004·Transplant Immunology·Ammon B Peck, Vijayakumar Ramiya
Mar 17, 2004·Experimental Cell Research·Alessandra CalabresePaolo Meda
Nov 16, 2004·Molecular and Cellular Endocrinology·Hans E Hohmeier, Christopher B Newgard
Oct 7, 2003·Annual Review of Biomedical Engineering·Martin L Yarmush, Scott Banta
Jun 2, 2015·Molecular Therapy. Methods & Clinical Development·Janet LawandiAnn M Simpson
Jul 13, 2000·Seminars in Veterinary Medicine and Surgery (small Animal)·D M Levitan
Aug 16, 2017·Nature Reviews. Gastroenterology & Hepatology·Cara EllisTimothy J Kieffer
Jun 25, 2016·The Biochemical Journal·Sanghamitra SinghAleksandar M Jeremic
Feb 27, 2001·Molecular Endocrinology·D de la TourF Levine
Aug 6, 2015·Archives of Pharmacal Research·Yue ZhuMing-Wei Wang
Jan 19, 2002·Annals of the New York Academy of Sciences·I ConstantinidisA Sambanis
Dec 14, 1999·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·J C Jiménez-ChillarónA M Gómez-Foix
Aug 30, 2013·The Journal of Biological Chemistry·Aurélien MichauMaude Le Gall
Nov 23, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jing HeRen-Bing Shi

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adenoma, Islet Cell

Islet Cell Adenoma arises in the islet cells, which are insulin producing cells of the pancreas. These tumors can be either malignant or benign. Discover the latest research on Islet Cell Adenoma here.

American Diabetes Association Journals

Discover the latest diabetes research published by the journals from the American Diabetes Association.