A site-specific genomic integration strategy for sustained expression of glucagon-like peptide-1 in mouse muscle for controlling energy homeostasis

Biochem Biophys Res Commun. 2010 Dec 10;403(2):172-7. doi: 10.1016/j.bbrc.2010.10.131. Epub 2010 Nov 9.

Abstract

The incretin hormone glucagon-like peptide-1 (GLP-1) exerts important functions in controlling glucose and energy homeostasis. Endogenous GLP-1 has a very short half-life due to DPP-IV-mediated degradation and renal clearance, which limits the therapeutic use of native GLP-1. We have shown previously that immunoglobulin fragment-fused GLP-1 (GLP-1/Fc) is a structurally stable GLP-1 analog. Here, we report a non-viral GLP-1/Fc gene therapy strategy utilizing a REP78-in-trans and REB-in-cis element system to achieve a site-specific genomic integration. For this purpose, the GLP-1/Fc expression cassette, which is fused with the RBE element, was co-injected with the Rep78 plasmid into the muscles of transgenic mice carrying the AAVS1 locus of human chromosome 19. The Rep protein-mediated site-specific integration was demonstrated by nested PCR, dot-blot, and Southern blotting. We found that this approach reduced weight gain and improved lipid profiles in the AAVS1-mice on high-fat diet challenge. Our observations reveal a new GLP-1 therapeutic strategy with an apparent absence of side effects, which may find applications in diabetes treatment and obesity prevention.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromosomes, Human, Pair 19 / genetics
  • DNA-Binding Proteins
  • Energy Metabolism / genetics
  • Genetic Engineering
  • Genetic Loci
  • Genetic Therapy / methods*
  • Genome
  • Glucagon-Like Peptide 1 / genetics*
  • Homeostasis / genetics
  • Humans
  • Mice
  • Mice, Transgenic
  • Muscle, Skeletal / metabolism*
  • Plasmids / genetics
  • Recombination, Genetic
  • Viral Proteins
  • Weight Gain / genetics*

Substances

  • DNA-Binding Proteins
  • Viral Proteins
  • rep proteins, Adeno-associated virus 2
  • Glucagon-Like Peptide 1