E-Selectin/AAV2/2 Gene Therapy Alters Angiogenesis and Inflammatory Gene Profiles in Mouse Gangrene Model

Abstract

For patients with chronic limb-threatening ischemia and limited revascularization options, alternate means for therapeutic angiogenesis and limb salvage are needed. E-selectin is a cell adhesion molecule that is critical for inflammation and neovascularization in areas of wound healing and ischemia. Here, we tested the efficacy of modifying ischemic limb tissue by intramuscular administration of E-selectin/AAV2/2 (adeno-associated virus serotype 2/2) to modulate angiogenic and inflammatory responses in a murine hindlimb gangrene model. Limb appearance, reperfusion, and functional recovery were assessed for 3 weeks after induction of ischemia. Mice receiving E-selectin/AAV2/2 gene therapy had reduced gangrene severity, increased limb and footpad perfusion, enhanced recruitment of endothelial progenitor cells, and improved performance on treadmill testing compared to control group. Histologically, E-selectin/AAV2/2 gene therapy was associated with increased vascularity and preserved myofiber integrity. E-selectin/AAV2/2 gene therapy also upregulated a panel of pro-angiogenic genes yet downregulated another group of genes associated with the inflammatory response. This novel gene therapy did not induce adverse effects on coagulability, or hematologic, hepatic, and renal function. Our findings highlight the potential of E-selectin/AAV2/2 gene therapy for improving limb perfusion and function in patients with chronic limb-threatening ischemia.

Bioinformatics Analyzes of Selectins Polymorphisms Reveal New Potential Cancer Biomarkers

“…Furthermore, when antitumor drugs were applied, gene expression levels were reduced, suggesting a possible role of E-selectin as an oncogene. Targeting E-selectin has emerged as a potential therapeutic strategy for the treatment of cancer [72][73][74]. Several approaches have been developed to target E-selectin, including monoclonal antibodies and small molecule inhibitors and have shown promise in preclinical studies by reducing tumor growth and metastasis [75][76][77][78][79].…”

Novel Gene-Modified Mesenchymal Stem Cell Therapy Reverses Impaired Wound Healing in Ischemic Limbs

“…A mouse model of hindlimb ischemia was utilized as described 23,24 . Briefly, 8- to 10-week-old male and female (50%; 50%) FVB/NJ mice (001800; Jackson Laboratory) underwent femoral artery and vein ligation at proximal and distal sites.…”

“…Briefly, 8- to 10-week-old male and female (50%; 50%) FVB/NJ mice (001800; Jackson Laboratory) underwent femoral artery and vein ligation at proximal and distal sites. Hindlimb ischemia was confirmed by preoperative and immediate postoperative laser Doppler imaging measurements were obtained as described 23–25 . Any mouse that did not demonstrate postoperative perfusion index of 5% or less of preoperative levels were excluded from further analysis in the study.…”

“…Hindlimb ischemia was confirmed by preoperative and immediate postoperative laser Doppler imaging measurements were obtained as described. [23][24][25] Any mouse that did not demonstrate postoperative perfusion index of 5% or less of preoperative levels were excluded from further analysis in the study. A cutaneous wound (4 mm punch biopsy) was created over the hindlimb ventral aspect.…”

Gene Therapeutic Strategies for Peripheral Artery Disease and New Opportunities Provided by Adeno-Associated Virus Vectors

“…Some of these are listed in Table 3. For example, Ribieras et al 152 found that intramuscular delivery of AAV2-E-selectin improved limb salvage, reperfusion, and function in a mouse hind limb gangrene model. E-selectin is a cell-adhesion molecule that mediates recruitment of the stem/progenitor cells that promote angiogenesis in ischemic tissue.…”