The Efficacy of HGF/VEGF Gene Therapy for Limb Ischemia in Mice with Impaired Glucose Tolerance: Shift from Angiogenesis to Axonal Growth and Oxidative Potential in Skeletal Muscle.

Autor: Stafeev I IS; National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, Russia., Boldyreva MA; National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, Russia.; Cell and Molecular Biotechnology Unit, Faculty of Biology and Biotechnology, National Research University Higher School of Economics, 101000 Moscow, Russia., Michurina SS; National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, Russia.; Faculty of Biology, Lomonosov Moscow State University, 117192 Moscow, Russia., Agareva MY; National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, Russia.; Institute of Fine Chemical Technologies Named after M.V. Lomonosov, 119571 Moscow, Russia., Radnaeva AV; Faculty of Medicine, Lomonosov Moscow State University, 119192 Moscow, Russia.; Institute for Regenerative Medicine, Medical Research and Education Centre, Lomonosov Moscow State University, 119192 Moscow, Russia., Menshikov MY; National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, Russia., Hu YC; Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300044, Taiwan.; Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University, Hsinchu 300044, Taiwan., Makarevich PI; Faculty of Medicine, Lomonosov Moscow State University, 119192 Moscow, Russia.; Institute for Regenerative Medicine, Medical Research and Education Centre, Lomonosov Moscow State University, 119192 Moscow, Russia., Parfyonova YV; National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, Russia.; Institute for Regenerative Medicine, Medical Research and Education Centre, Lomonosov Moscow State University, 119192 Moscow, Russia.
Jazyk: angličtina
Zdroj: Cells [Cells] 2022 Nov 29; Vol. 11 (23). Date of Electronic Publication: 2022 Nov 29.
DOI: 10.3390/cells11233824
Abstrakt: Background: Combined non-viral gene therapy (GT) of ischemia and cardiovascular disease is a promising tool for potential clinical translation. In previous studies our group has developed combined gene therapy by vascular endothelial growth factor 165 ( VEGF165 ) + hepatocyte growth factor ( HGF ). Our recent works have demonstrated that a bicistronic pDNA that carries both human HGF and VEGF165 coding sequences has a potential for clinical application in peripheral artery disease (PAD). The present study aimed to test HGF/VEGF combined plasmid efficacy in ischemic skeletal muscle comorbid with predominant complications of PAD-impaired glucose tolerance and type 2 diabetes mellitus (T2DM).
Methods: Male C57BL mice were housed on low-fat (LFD) or high-fat diet (HFD) for 10 weeks and metabolic parameters including FBG level, ITT, and GTT were evaluated. Hindlimb ischemia induction and plasmid administration were performed at 10 weeks with 3 weeks for post-surgical follow-up. Limb blood flow was assessed by laser Doppler scanning at 7, 14, and 21 days after ischemia induction. The necrotic area of m.tibialis anterior , macrophage infiltration, angio- and neuritogenesis were evaluated in tissue sections. The mitochondrial status of skeletal muscle (total mitochondria content, ETC proteins content) was assessed by Western blotting of muscle lysates.
Results: At 10 weeks, the HFD group demonstrated impaired glucose tolerance in comparison with the LFD group. HGF/VEGF plasmid injection aggravated glucose intolerance in HFD conditions. Blood flow recovery was not changed by HGF/VEGF plasmid injection either in LFD or HFD conditions. GT in LFD, but not in HFD conditions, enlarged the necrotic area and CD68+ cells infiltration. However, HGF/VEGF plasmid enhanced neuritogenesis and enlarged NF200+ area on muscle sections. In HFD conditions, HGF/VEGF plasmid injection significantly increased mitochondria content and ETC proteins content.
Conclusions: The current study demonstrated a significant role of dietary conditions in pre-clinical testing of non-viral GT drugs. HGF/VEGF combined plasmid demonstrated a novel aspect of potential participation in ischemic skeletal muscle regeneration, through regulation of innervation and bioenergetics of muscle. The obtained results made HGF/VEGF combined plasmid a very promising tool for PAD therapy in impaired glucose tolerance conditions.
Databáze: MEDLINE
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