Effects of adenoviruses carrying genes of a vascular endothelial growth factor (vegf 165), a glial-cell derived neurotrophic factor (gdnf), and angiogenin (ang) were studied in a rat model of chronic hindlimb ischemia. Therapeutic genes were used for direct gene therapy in the following combinations: (1) Ad5-ANG; (2) Ad5-VEGF+Ad5-ANG; and (3) Ad5-VEGF+Ad5-ANG+Ad5-GDNF. Real-time PCR demonstrated increased gdnf, vegf, and ang mRNA levels within the area of an ischemic muscle on day 14 where the study combinations of therapeutic genes were injected in both Ad5-VEGF+Ad5-ANG and Ad5-VEGF+Ad5-ANG+Ad5-GDNF groups. On post-injection day 28, the number of centronuclear myotubes (CNMs) as well as the CD31-immunopositive cell count showed a 38.7- and 1.3-fold increase, respectively, within the ischemic area in the Ad5-VEGF+Ad5-ANG+Ad5-GDNF group compared with the Ad5-VEGF+Ad5-ANG group. There was an increased expression of desmin mRNA in the area of an ischemic muscle in Ad5-VEGF+Ad5-ANG and Ad5-VEGF+Ad5-ANG+Ad5-GDNF groups on day 14. Based on Western blotting results, the expression of CD34 and a von Willebrand factor (VWF) increased on day 14 after injection of Ad5-VEGF+Ad5-ANG+Ad5-GDNF as compared with the control group. The Ad5-VEGF+Ad5-ANG+Ad5-GDNF injection stimulates muscle regeneration by increasing the number of CNMs and blood vessels. Based on the above findings, the gdnf angiogenic and regenerative potential necessitates further studies of its possible use as an agent stimulating angiogenesis and skeletal muscle regeneration for clinical purposes.
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Fowkes, G., Rudan, D., Rudan, I., & Aboyans, V. (2013). Comparison of global estimates of prevalence and risk factors for peripheral artery disease in 2000 and 2010: a systematic review and analysis. Lancet, 382, 1329–1340. https://doi.org/10.1016/S0140-6736(13)61249-0.
Guo, S., Colbert, L., Fuller, M., Zhang, Y., & Gonzalez-Perez, R. (2010). Vascular endothelial growth factor receptor2 in breast cancer. Biochimica et Biophysica Acta, 1806(1), 108–121. https://doi.org/10.1016/j.bbcan.2010.04.004.
Jazwa, A., Tomczyk, M., Taha, M., Hytonen, E., Stoszko, M., Zentilin, L., Giacca, M., Yla-Herttuala, S., Emanueli, C., Jozkowicz, A., & Dulak, J. (2013a). Arteriogenic therapy based on simultaneous delivery of VEGF-A and FGF4 genes improves the recovery from acute limb ischemia. Vascular Cell, 5–13. https://doi.org/10.1186/2045-824X-5-13.
Jazwa, A., Tomczyk, M., Taha, M., Hytonen, E., et al. (2013b). Arteriogenic therapy based on simultaneous delivery of VEGF-A and FGF4 genes improves the recovery from acute limb ischemia. Vascular Cell, 2013, 5–13. https://doi.org/10.1186/2045-824X-5-13.
Boden, J., Lassance-Soares, R., Wang, H., Wei, Y., & Spiga, M. (2016). Vascular regeneration in ischemic hindlimb by adeno-associated virus expressing conditionally silenced vascular endothelial growth factor. Journal of the American Heart Association, 110–119. https://doi.org/10.1161/JAHA.115.001815.
Sanada, F., Taniyama, Y., Azuma, J., Yuka, I., Kanbara, Y., Iwabayashi, M., Rakugi, H., & Morishita, R. (2014). Therapeutic angiogenesis by gene therapy for critical limb ischemia: choice of biological agent. Immunology, Endocrine & Metabolic Agents in Medicinal Chemistry, 14(1), 32–39. https://doi.org/10.2174/1871522213999131231105139.
Shapiro, R., Riordan, F., & Vallee, L. (1986). Characteristic ribonucleolytic activity of human angiogenin. Biochemistry, 25, 3527–3532. https://doi.org/10.1021/bi00360a008.
Gavrilenko, V., Voronov, A., Konstantinov, A., & Bochkov, P. (2008). Combination of reconstructive vascular operations with gene-engineering technologies of angiogenesis stimulation: a present-day policy aimed at improving the remote results of treating patients with lower limb chronic ischaemia. Angiol Sosud Khir, 14(4), 49–53.
Konstantinov, B., Bochkov, P., & Gavrilenko, V. (2003). Opportunity and the prospects of treatment of critical ischemia with use of genetically engineered technologies. Angiology and Vascular Surgery, 9(3), 14–18.
Verkhovskaya, V., Sheremetyeva, F., Gavrilenko, G., & Tarantul, Z. (2004). Infinity biological functions and therapeutic properties of angiogenin. Molecular Genetics, Microbiology and Virusology, 4, 38–40.
Zhonghua Y, Xue Z (2015). Effects of denervation on angiogenesis and skeletal muscle fiber remodeling of ischemic limbs. 95(8): 601-5. https://doi.org/10.3760/cma.j.issn.0376-2491.2015.08.010
Cen, Y., Liu, J., Qin, Y., Liu, R., Wang, H., Zhou, Y., Wang, S., & Hu, Z. (2016). Denervation in femoral artery-ligated hindlimbs diminishes ischemic recovery primarily via impaired arteriogenesis. PLoS One, 11(5), 50–62. https://doi.org/10.1371/journal.pone.0154941.
Zhang, R., Lu, Y., Li, J., Wang, J., Liu, C., Gao, F., & Sun, D. (2016). Glial cell line-derived neurotrophic factor induced the differentiation of amniotic fluid-derived stem cells into vascular endothelial-like cells in vitro. Journal of Molecular Histology, 47(1), 9–19. https://doi.org/10.1007/s10735-015-9649-9.
Kingham, J., Kolar, K., Novikova, N., Novikov, N., & Wiberg, M. (2013). Stimulating the neurotrophic and angiogenic properties of human adipose-derived stem cells enhances nerve repair. Stem Cells and Development, 23(7), 741–754. https://doi.org/10.1089/scd.2013.0396.
Mukhamedshina, Y., Shaymardanova, G., Garanina, E., et al. (2016). Adenoviral vector carrying glial cell-derived neurotrophic factor for direct gene therapy in comparison with human umbilical cord blood cell-mediated therapy of spinal cord injury in rat. Spinal Cord, 54(5), 347–359. https://doi.org/10.1038/sc.2015.161.
Mukhamedshina, Y., Akhmetzyanova, E., Kostennikov, A., Zakirova, E., Galieva, L., Garanina, E., Rogozin, A., Kiyasov, A., & Rizvanov, A. (2018). Adipose-derived mesenchymal stem cells application combined with fibrin matrix promote structural and functional recovery following spinal cord injury in rats. Frontiers in Pharmacology, 9, 343. https://doi.org/10.4103/1673-5374.244778.
Makarevich, P., Dergilev, K., Tsokolaeva, Z., Boldyreva, M., Shevchenko, E., Gluhanyuk, E., Gallinger, J., Menshikov, M., & Parfyonova, Y. (2018). Angiogenic and pleiotropic effects of VEGF165 and HGF combined gene therapy in a rat model of myocardial infarction. PLoS One, 13(5), 1–25. https://doi.org/10.1371/journal.pone.0197566.
Blais, M., Lévesque, P., Bellenfant, S., & Berthod, F. (2013). Nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3 and glial-derived neurotrophic factor enhance angiogenesis in a tissue-engineered in vitro model. Tissue Engineering Part A, 19(15-16), 1655–1664. https://doi.org/10.1089/ten.tea.2012.0745.
Titova A, Mavlikeev M, Pevnev G, Bilyalov A, Abyzova M. et al. (2017) Histological assessment of pathological changes in the calf muscle in the simulation of hind limb ischemia in Kazan medical journal. 2017. 98: 73-76.
Boldyreva, M., Makarevich, P., Rafieva, L., Beloglazova, I., Gergiev, K., Kostrov, S., & Parfenov, E. (2014). Gene therapy based on recombinant plasmid with the gene of nerve growth factor (ngf) stimulates angiogenesis and restoration of blood flow in the ischemic hind limb of the mouse. Genes to Cells, IX(4), 81–87.
Yong, P., Feng, K., Sheng, Y., Zuo-Guan, C., Li-Shan, L., Li-Long, G., & Yong, J. (2016). Nerve growth factor promotes angiogenesis and skeletal muscle fiber remodeling in a murine model of hindlimb ischemia. Chinese Medical Journal, 129(3), 95–110. https://doi.org/10.4103/0366-6999.174496.
Shvartsman, D., Storrie-White, H., Kangwon, L., et al. (2014). Sustained delivery of VEGF maintains innervation and promotes reperfusion in ischemic skeletal muscles via NGF/GDNF signaling. Molecular Therapy, 22(7), 1243–1253. https://doi.org/10.1038/mt.2014.76.
The authors acknowledge their gratitude to M.M. Shmarov, B.S. Naroditskyi, R.R. Islamov, and D.R. Galieva.
This study was supported by grant NSC-3076.2018.4 (AR). This work was performed in accordance with the Program of Competitive Growth of the Kazan Federal University. AR was supported by a state assignment 20.5175.2017/6.7 (Leading Researcher) of the Ministry of Education and Science of the Russian Federation.
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Samatoshenkov, I.V., Salafutdinov, I.I., Zuravleva, M.N. et al. Adenoviral Vector Delivery of vegf, Angiogenin, and gdnf Genes Promotes Angiogenesis in Ischemic Skeletal Muscle. BioNanoSci. (2020). https://doi.org/10.1007/s12668-019-00688-y
- Limb ischemia