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Insulin delivery with plasmid DNA.

A M Abai¹, P M Hobart, K M Barnhart

¹Department of Molecular Biology, Vical, Inc., San Diego, CA 92121, USA. aabai@vical.com

Human Gene Therapy

|November 24, 1999

Summary

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Restoring basal insulin levels through plasmid DNA (pDNA) injection into muscle shows promise for type I diabetes. This approach successfully produced biologically active insulin in mice, improving survival rates in diabetic models.

Area of Science:

Molecular Biology
Endocrinology
Gene Therapy

Background:

Controlling hyperglycemia in type I diabetes necessitates basal insulin restoration.
Plasmid DNA (pDNA) offers a potential vehicle for therapeutic insulin delivery.

Purpose of the Study:

To evaluate the efficacy of pDNA encoding preproinsulin for constitutive expression and processing into mature insulin in nonendocrine cells.
To assess the in vivo therapeutic potential of skeletal muscle-mediated insulin production for basal insulin delivery.

Main Methods:

Comparison of three pDNAs expressing modified rat proinsulin I in vitro.
In vivo intramuscular injection of pDNA in mice to assess proinsulin expression and serum levels.
Evaluation of therapeutic efficacy in streptozotocin-induced diabetic mice using pDNA encoding modified proinsulin and furin.

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Main Results:

A modified proinsulin pDNA (VR-3503) with a furin cleavage site yielded 30-70% mature insulin in vitro, increasing to >99% with cotransfection of a furin expression plasmid.
Intramuscular injection of pDNA resulted in sustained serum proinsulin levels in mice, particularly in immune-compromised models.
Coinjection of VR-3503 and VR-3505 pDNAs in diabetic mice significantly improved survival rates, with surviving animals showing substantial levels of mature, immune-reactive insulin in muscle.

Conclusions:

Skeletal muscle can be engineered to express and deliver biologically active, mature insulin from plasmid DNA.
This gene therapy approach holds potential as a valid strategy for basal insulin replacement in type I diabetes.