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Glucose-stimulated and self-limiting insulin production by glucose 6-phosphatase promoter driven insulin expression in hepatoma cells

Abstract

The liver is an attractive target organ for insulin gene expression in type 1 diabetes as it contains appropriate cellular mechanisms of regulated gene expression in response to blood glucose and insulin. We hypothesize that insulin production regulated by both glucose and insulin may be achieved using the promoter of the glucose 6-phosphatase gene (G6Pase), the expression of which in the liver is induced by glucose and suppressed by insulin. Recombinant adenoviral vectors expressing the reporter gene CAT or insulin under transcriptional direction of the G6Pase promoter were constructed. Glucose-stimulated as well as self-limiting insulin production was achieved in vector-transduced hepatoma cells in which expression of the insulin gene was controlled by the G6Pase promoter. While insulin strongly inhibited the G6Pase promoter activity under low glucose conditions, its inhibitory capacity was attenuated when glucose levels were elevated. At the physiologic glucose level of 5.5 mM glucose, vector-transduced hepatoma cells produced a self-limited level of insulin at approximately 0.2–0.3 ng/ml, which is within the range of fasting levels of insulin in normal animals. These results indicate that the G6Pase promoter possesses desirable features and may be developed for regulated hepatic insulin gene expression in type 1 diabetes.

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Acknowledgements

We thank Drs Nuria Morral and Hengjiang Dong for critical review of this manuscript. We thank Oxana Galenko for her assistance in rescuing recombinant adenoviruses, and Jennifer Altomonte and Liqun Wang for their assistance in the measurement of insulin and glucose. This work was supported in part by NIH grant DK54377 and by the JDFI center at Mount Sinai School of Medicine. R Chen was supported by a Juvenile Diabetes Foundation International Fellowship (JDFI-398232).

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Chen, R., Meseck, M., McEvoy, R. et al. Glucose-stimulated and self-limiting insulin production by glucose 6-phosphatase promoter driven insulin expression in hepatoma cells. Gene Ther 7, 1802–1809 (2000). https://doi.org/10.1038/sj.gt.3301306

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  • DOI: https://doi.org/10.1038/sj.gt.3301306

Keywords

  • glucose-6-phosphatase promoter
  • insulin
  • glucose
  • regulated insulin gene expression
  • type 1 diabetes

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