title: Pluronic L64-mediated stable HIF-1α expression in muscle for therapeutic angiogenesis in mouse hindlimb ischemia.
abstract: Intramuscular injection of plasmid DNA (pDNA) to express a therapeutic protein is a promising method for the treatment of many diseases. However, the therapeutic applications are usually hindered by gene delivery efficiency and expression level. In this study, critical factors in a pDNA-based gene therapy system, such as gene delivery materials, a therapeutic gene, and its regulatory elements, were optimized to establish an integrated system for the treatment of mouse hindlimb ischemia. The results showed that Pluronic L64 (L64) was an efficient and safe material for gene delivery into mouse skeletal muscle. It also showed intrinsic ability to promote in vivo angiogenesis in a concentration-dependent manner, which might be through the activation of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB)-regulated angiogenic factors. The combination of 0.1% L64 with a hybrid gene promoter (pSC) increased the gene expression level, elongated the gene expression duration, and enhanced the number of transfected muscle fibers. In mice ischemic limbs, a gene medicine (pSC-HIF1α(tri)/L64) composed of L64 and pSC-based expression plasmid encoding hypoxia-inducible factor 1-alpha triple mutant (HIF-1α(tri)), improved the expression of stable HIF-1α, and in turn, the expression of multiple angiogenic factors. As a result, the ischemic limbs showed accelerated function recovery, reduced foot necrosis, faster blood reperfusion, and higher capillary density. These results indicated that the pSC-HIF1α(tri)/L64 combination presented a potential and convenient venue for the treatment of peripheral vascular diseases, especially critical limb ischemia.
authors.affiliation: National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, People's Republic of China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Song
authors.affiliation: National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, People's Republic of China.
authors.forename: Sijia
authors.initials: S
authors.lastname: Liu
authors.affiliation: National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, People's Republic of China ; Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, People's Republic of China.
authors.forename: Caixia
authors.initials: C
authors.lastname: Li
authors.affiliation: National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, People's Republic of China ; Hebei University of Engineering, Handan, Hebei, People's Republic of China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Geng
authors.affiliation: National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, People's Republic of China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.affiliation: National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, People's Republic of China.
authors.forename: Zhongwei
authors.initials: Z
authors.lastname: Gu
mesh_terms: D043925:Angiogenesis Inducing Agents; D000818:Animals; D002460:Cell Line; D015316:Genetic Therapy / Q000379:methods*; D006614:Hindlimb / Q000293:injuries; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D007511:Ischemia / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000235:genetics*; D020442:Poloxamer / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
chemical_list: D043925:Angiogenesis Inducing Agents; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016328:NF-kappa B; D020442:Poloxamer; C514452:pluronic L64
keywords: HIF-1α; Pluronic L64; SV40 enhancer; angiogenesis; gene therapy
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