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
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 (<i>VEGF165</i>) + hepatocyte growth...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-11-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/11/23/3824 |
| _version_ | 1827643283444072448 |
|---|---|
| author | Iurii S. Stafeev I Maria A. Boldyreva Svetlana S. Michurina Margarita Yu. Agareva Arina V. Radnaeva Mikhail Yu. Menshikov Yu-Chen Hu Pavel I. Makarevich Yelena V. Parfyonova |
| author_facet | Iurii S. Stafeev I Maria A. Boldyreva Svetlana S. Michurina Margarita Yu. Agareva Arina V. Radnaeva Mikhail Yu. Menshikov Yu-Chen Hu Pavel I. Makarevich Yelena V. Parfyonova |
| author_sort | Iurii S. Stafeev I |
| collection | DOAJ |
| description | 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 (<i>VEGF165</i>) + hepatocyte growth factor (<i>HGF</i>). Our recent works have demonstrated that a bicistronic pDNA that carries both human <i>HGF</i> and <i>VEGF165</i> coding sequences has a potential for clinical application in peripheral artery disease (PAD). The present study aimed to test <i>HGF/VEGF</i> 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 <i>m.tibialis anterior</i>, 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. <i>HGF/VEGF</i> plasmid injection aggravated glucose intolerance in HFD conditions. Blood flow recovery was not changed by <i>HGF/VEGF</i> 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, <i>HGF/VEGF</i> plasmid enhanced neuritogenesis and enlarged NF200+ area on muscle sections. In HFD conditions, <i>HGF/VEGF</i> 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. <i>HGF/VEGF</i> 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 <i>HGF/VEGF</i> combined plasmid a very promising tool for PAD therapy in impaired glucose tolerance conditions. |
| first_indexed | 2024-03-09T17:51:18Z |
| format | Article |
| id | doaj.art-58b2e8ff11524bf08842ac3272e091eb |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T17:51:18Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-58b2e8ff11524bf08842ac3272e091eb2023-11-24T10:44:23ZengMDPI AGCells2073-44092022-11-011123382410.3390/cells11233824The 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 MuscleIurii S. Stafeev I0Maria A. Boldyreva1Svetlana S. Michurina2Margarita Yu. Agareva3Arina V. Radnaeva4Mikhail Yu. Menshikov5Yu-Chen Hu6Pavel I. Makarevich7Yelena V. Parfyonova8National Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, RussiaNational Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, RussiaNational Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, RussiaNational Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, RussiaFaculty of Medicine, Lomonosov Moscow State University, 119192 Moscow, RussiaNational Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, RussiaDepartment of Chemical Engineering, National Tsing Hua University, Hsinchu 300044, TaiwanFaculty of Medicine, Lomonosov Moscow State University, 119192 Moscow, RussiaNational Medical Research Centre for Cardiology Named after Academician E.I.Chazov, 121552 Moscow, RussiaBackground: 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 (<i>VEGF165</i>) + hepatocyte growth factor (<i>HGF</i>). Our recent works have demonstrated that a bicistronic pDNA that carries both human <i>HGF</i> and <i>VEGF165</i> coding sequences has a potential for clinical application in peripheral artery disease (PAD). The present study aimed to test <i>HGF/VEGF</i> 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 <i>m.tibialis anterior</i>, 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. <i>HGF/VEGF</i> plasmid injection aggravated glucose intolerance in HFD conditions. Blood flow recovery was not changed by <i>HGF/VEGF</i> 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, <i>HGF/VEGF</i> plasmid enhanced neuritogenesis and enlarged NF200+ area on muscle sections. In HFD conditions, <i>HGF/VEGF</i> 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. <i>HGF/VEGF</i> 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 <i>HGF/VEGF</i> combined plasmid a very promising tool for PAD therapy in impaired glucose tolerance conditions.https://www.mdpi.com/2073-4409/11/23/3824gene therapyplasmidlimb ischemiahigh-fat dietobesityVEGF |
| spellingShingle | Iurii S. Stafeev I Maria A. Boldyreva Svetlana S. Michurina Margarita Yu. Agareva Arina V. Radnaeva Mikhail Yu. Menshikov Yu-Chen Hu Pavel I. Makarevich Yelena V. Parfyonova 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 Cells gene therapy plasmid limb ischemia high-fat diet obesity VEGF |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | gene therapy plasmid limb ischemia high-fat diet obesity VEGF |
| url | https://www.mdpi.com/2073-4409/11/23/3824 |
| work_keys_str_mv | AT iuriisstafeevi theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT mariaaboldyreva theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT svetlanasmichurina theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT margaritayuagareva theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT arinavradnaeva theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT mikhailyumenshikov theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT yuchenhu theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT pavelimakarevich theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT yelenavparfyonova theefficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT iuriisstafeevi efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT mariaaboldyreva efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT svetlanasmichurina efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT margaritayuagareva efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT arinavradnaeva efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT mikhailyumenshikov efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT yuchenhu efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT pavelimakarevich efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle AT yelenavparfyonova efficacyofhgfvegfgenetherapyforlimbischemiainmicewithimpairedglucosetoleranceshiftfromangiogenesistoaxonalgrowthandoxidativepotentialinskeletalmuscle |