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

A. M. Simpson*, G. M. Marshall, B. E. Tuch, L. Maxwell, B. Szymanska, J. Tu, S. Beynon, Ma Swan, M. Camacho

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    36 Citations (Scopus)

    Abstract

    In order to design a feasible somatic cell gene delivery system for the treatment of type 1 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/gl cells). The HEP G2ins/g cell clones exhibit glucose-stimulated insulin secreted 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.

    Original languageEnglish
    Pages (from-to)1202-1215
    Number of pages14
    JournalGene Therapy
    Volume4
    Issue number11
    Publication statusPublished - 1997

    Keywords

    • Electron microscopy
    • Gene therapy
    • Glucose transport protein
    • Insulin storage
    • Liver

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