Abstract 10451: A Potential Therapeutic Antibody to Induce Collateral Formation in Cardiovascular Diseases

• Published in: Circulation (New York, N.Y.) N.Y.), 2021-11, Vol.144 (Suppl_1), p.A10451-A10451
• Main Authors: Amartey, Jason, Wahid, Mussarat, Bhalla, Sohni R, Bates, David O
• Format: Article
• Language: English
• ISSN: 0009-7322; 1524-4539
• DOI: 10.1161/circ.144.suppl_1.10451

IntroductionLack of efficient collateralisation during vascular occlusion leads to tissue ischemia in coronary, peripheral and cerebrovascular disease. Patients with peripheral, coronary, and cerebrovascular disease have elevated levels of the anti-angiogenic isoform of VEGF (VEGF-A165b), which correlate with poorer outcomes. In experimental animals, VEGF-A165b neutralisation improves blood flow and collateralisation after femoral or coronary artery ligation. HypothesisWe tested the hypothesis that a potent, high affinity, humanised neutralising antibody to VEGF-A165b could have therapeutic potential in models of cardiovascular disease. MethodsA mouse monoclonal antibody to VEGF-A165b was humanised by cloning and site directed mutagenesis. Multiple clones were screened, and one with high affinity (measured by biolayer interferometry) was amplified. The efficacy in vitro was tested using endothelial cell migration assays and in vivo using the hindlimb ischemia (HLI) model in C57Bl6 mice fed a high fat high sucrose (HFHS) diet, and in Zucker Diabetic Fatty rats. ResultsOne clone (HC4LC2) had a VEGF-A165b affinity of 600pM. 0.3μg/ml of this clone reversed the inhibitory effect of recombinant human VEGF-A165b (31±4%, N=6 of VEGF-A165a alone) on migration of endothelial cells to 40ng/ml rhVEGF-A165a (89±9% N=3). Human monocyte dependent inhibition of VEGF-A165a mediated migration was dose dependently reversed by HC4LC2 (EC50, 55ng/ml, N=9 per concentration). HLI in HFHS mice impaired blood flow after 28 days in IgG treated mice (70±7.7% of pre-ischemic blood flow, N=6), which was restored to normal (93.3±5.5, p