Glucose-regulated insulin production from genetically engineered human non-beta cells.
In this report we describe development and characterization of four human cell lines that are able to secrete insulin and C-peptide in response to higher concentrations of glucose. These cell lines have been developed by stably and constitutively expressing human proinsulin with a furin-cleavable site, whereas expression of furin is regulated by glucose concentration. These cell lines have been cloned and, therefore, the transgene in each cell is located in an identical location of the genome leading to a uniform expression. Cloning has also allowed us to identify cell lines with more desirable properties such as higher basal insulin secretion and/or better glucose responsiveness. We have further shown that the insulin produced by these cells is biologically active and induces normoglycemia when injected in diabetic animals. Our objective in initiating these studies was to identify a cell line that could serve as a surrogate beta cell line for therapeutic intervention in type I diabetic patients.
Tatake
Revati J
RJ
Boehringer Ingelheim Pharmaceuticals, 900 Ridgebury Road, Ridgefield, CT 06877, USA. rtatake@rdg.boehringer-ingelheim.com
O'Neill
Margaret M
MM
Kennedy
Charles A
CA
Reale
Virginia D
VD
Runyan
Jacob D
JD
Monaco
Kelli-Ann D
KA
Yu
Kyung
K
Osborne
William R
WR
Barton
Randall W
RW
Schneiderman
Richard D
RD
D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D002096:C-Peptide / Q000378:metabolism; D002460:Cell Line; D003470:Culture Media; D018076:DNA, Complementary / Q000235:genetics; D003921:Diabetes Mellitus, Experimental / Q000188:drug therapy / Q000378:metabolism; D005818:Genetic Engineering; D005822:Genetic Vectors; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism* / Q000627:therapeutic use*; D000078790:Insulin Secretion; D007515:Islets of Langerhans; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010957:Plasmids; D011384:Proinsulin / Q000235:genetics; D011401:Promoter Regions, Genetic; D051381:Rats; D011923:Rats, Nude; D011972:Receptor, Insulin / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014162:Transfection
D001786:Blood Glucose; D002096:C-Peptide; D003470:Culture Media; D018076:DNA, Complementary; D007328:Insulin; D011384:Proinsulin; D017853:Phosphotransferases (Alcohol Group Acceptor); C023916:phosphoglycerol kinase; D011972:Receptor, Insulin; D005947:Glucose