| Autor: |
Voza, Francesca A., Byrne, Barry J., Ortiz, Yulexi Y., Yan Li, Nga Le, Osafo, Lucy, Ribieras, Antoine C., Shao, Hongwei, Huerta, Carlos Theodore, Yuntao Wei, Falero-Diaz, Gustavo, Franco-Bravo, Andres, Lassance-Soares, Roberta M., Vazquez-Padron, Roberto I., Zhao-Jun Liu, Velazquez, Omaida C. |
| Zdroj: |
Annals of Surgery; Oct2024, Vol. 280 Issue 4, p570-583, 14p |
| Abstrakt: |
Objective: This study focuses on dose--response investigation using a codon-optimized and de novo--synthesized E-Selectin/AAV2 (E-Sel/AAV2) vector in preparation for Investigational New Drug enabling of subsequent clinical studies. Background: Gene therapy is a potential solution for patients suffering from chronic limb-threatening ischemia. Understanding the dose for effective gene delivery is crucial for future Investigational New Drug--enabling studies. Methods: Expression of the codon-optimized E-Selectin gene was assessed by flow cytometry following in vitro cell transfection assay and RT-qPCR for murine limbs injected in vivo with AAV-m-ESelectin (E-Sel/AAV2). Dose--response studies involved 3 cohorts of FVB/NJ mice (n=6/group) with escalating log doses of E-Selectin/ AAV2 injected intramuscularly in divided aliquots, ranging from 2 x 109 VG to 2 x 1011 VG, into ischemic limbs created by left femoral artery/vein ligation/excision and administration of nitric oxide synthase inhibitor, L-NAME. Limb perfusion, extent of gangrene free limb, functional limb recovery, and therapeutic angiogenesis were assessed. Results: Codon-optimized E-Sel/AAV2 gene therapy exhibits a superior expression level than WT E-Sel/AAV2 gene therapy both in vitro and in vivo. Mice treated with a high dose (2 x 1011 VG) of E-Sel/AAV2 showed significantly improved perfusion indices, lower Faber scores, increased running stamina, and neovascularization compared with lower doses tested with control groups, indicating a distinct dose-dependent response. No toxicity was detected in any of the animal groups studied. Conclusions: E-Sel/AAV2 Vascular Regeneration Gene Therapy holds promise for enhancing the recovery of ischemic hindlimb perfusion and function, with the effective dose identified in this study as 2 x 1011 VG aliquots injected intramuscularly. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Supplemental Index |
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