A site-specific genomic integration strategy for sustained expression of glucagon-like peptide-1 in mouse muscle for controlling energy homeostasis.
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.
Department of Endocrinology, Hua Shan Hospital, Fudan University, Shanghai, China.
Rui
R
Liu
Yiming
Y
Li
Renmin
R
Hu
Tianru
T
Jin
Shanshan
S
Deng
Wang
W
Liang
Nina
N
Zhang
Jinzhong
J
Chen
Gerald J
GJ
Prud'homme
William W
WW
Jia
Duan
D
Ma
Qinghua
Q
Wang
D000818:Animals; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics; D004268:DNA-Binding Proteins; D004734:Energy Metabolism / Q000235:genetics; D005818:Genetic Engineering; D056426:Genetic Loci; D015316:Genetic Therapy / Q000379:methods*; D016678:Genome; D052216:Glucagon-Like Peptide 1 / Q000235:genetics*; D006706:Homeostasis / Q000235:genetics; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D014764:Viral Proteins; D015430:Weight Gain / Q000235:genetics*
D004268:DNA-Binding Proteins; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2; D052216:Glucagon-Like Peptide 1