In Silico designed cell-penetrating anti-cancer peptide specifically inhibits VEGF-A expression

Abstract

Vascular Endothelial Growth Factor A (VEGF A), a pluripotent cytokine and angiogenic growth factor mediates the switch to an angiogenic phenotype in cancer cells. The interaction of VEGF A protein with the VEGF receptors (VEGFR 1and VEGFR 2) starts downstream effect that promotes angiogenesis by mediating migration and increasing the permeability of endothelial cells. A cis regulatory elements consisting of a polypurine polypyrimidine tract in the proximal 36-bp region, can participate in the formation of a stable higher order G quadruplex structure (G4) which is essential for VEGF promoter activity. During cancer progression the VEGF A G4 succumbs to cellular pressure and fails to maintain the stable structure. This shifts the balance to form duplex structure thereby increasing the rate of transcription. Earlier research has tried to develop small molecule ligands to target and stabilize G4, however they either lack specificity or non toxicity. Peptide on the other hand are very less studied. Here we used bioinformatics in silico tool to develop peptides which can successfully bind and stabilize the VEGF A G4 while reducing its gene expression. This further alters the expression fate of the VEGF A signalling cascade and prevents angiogenesis in cancer cells. We used high resolution Nuclear magnetic resonance and molecular dynamics simulation to map the chemistry of the interaction while the qPCR and western blot allowed us to check the expression pattern of the molecules of VEGF A signalling cascade. In this investigation, we navigate the complex interplay between peptides and quadruplex structures, unravelling valuable insights that can enhance the crafting of pharmacophores directed at the dynamic quadruplex structure. The outcomes of our study are promising, paving the way for progress in the realms of research, characterization, and optimization of peptides binding to G quadruplexes, with potential implications for therapeutic applications.