Gene therapy of diabetes : glucose-stimulated insulin secretion in a human hepatoma cell line (HEP G2ins/g)

• Published in: Gene therapy Vol. 4; no. 11; pp. 1202 - 1215
• Main Authors: SIMPSON, A. M, MARSHALL, G. M, TUCH, B. E, MAXWELL, L, SZYMANSKA, B, TU, J, BEYNON, S, SWANS, M. A, CAMACHO, M
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.11.1997
• Subjects: Beta cells; Biological and medical sciences; Biotechnology; Blotting, Northern; Blotting, Western; Carcinoma, Hepatocellular; Cell Line; Diabetes; Diabetes mellitus; Diabetes Mellitus, Type 1 - therapy; Fundamental and applied biological sciences. Psychology; Gene therapy; Gene transfer; Genetic Engineering; Genetic Therapy - methods; Glucose; Glucose - pharmacology; Glucose transporter; Glucose Transporter Type 2; Health. Pharmaceutical industry; Hepatocytes; Hepatoma; Humans; Immunoelectron microscopy; Industrial applications and implications. Economical aspects; Insulin; Insulin - metabolism; Insulin Secretion; Islets of Langerhans - metabolism; Liver cancer; Monosaccharide Transport Proteins - analysis; Monosaccharide Transport Proteins - genetics; Physiology; Stimulation, Chemical; Transfection; Endocrinopathy; Human; Protein hormone; Cell line; Gene; Secretion; Hepatocellular carcinoma; Glucose; Recombinant cell; Gene therapy; Non insulin dependent diabetes; Insulin
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3300527

In order to design a feasible somatic cell gene delivery system for the treatment of type I diabetes, a suitable cell type needs to be determined. We have previously shown that the stable transfection of the full-length insulin cDNA into the human liver cell line, (HEP G2ins) resulted in synthesis, storage and acute regulated release of insulin to analogues of cAMP, but not to the physiological stimulus glucose. In attempting to explain the lack of glucose responsiveness of the HEP G2ins cells we have stably transfected these cells with the human islet glucose transporter GLUT 2 (HEP, G2ins/g cells). The HEP G2ins/g cell clones exhibit glucose-stimulated insulin secretion and glucose potentiation of the secretory response to nonglucose secretagogues. While glucose responsiveness commenced at a lower concentration than normal islets, a secretion curve approaching normal physiological conditions was generated. Immunoelectron microscopy revealed the presence of insulin-containing granules, similar in size and appearance to those of the normal beta cell. These results demonstrate that while it is most likely that the HEP G2ins/g cell line predominantly secretes insulin via the constitutive pathway, significant acute regulated release was seen in response to glucose, and thus represents significant progress in the creation of a genetically engineered 'artificial beta cell' from a human hepatocyte cell line.