Cascaded Bio-responsive Delivering eNOS Gene and ZNF580 Gene to Collaboratively Treat Hindlimb Ischemia via Pro-angiogenesis and Anti-inflammation
Gene therapy is a promising strategy for treating ischemic disease by solving the dual-dilemma of ischemia and inflammation. However, its development remains limited by inefficient gene transfection. Hence, we have proposed a “dual genes + all-adaptive carrier” idea. We have innovatively co-delivered eNOS gene and ZNF580 gene encoding its transcription factor for enhancing efficient eNOS expression. The overexpressed ZNF580 protein significantly promotes angiogenesis via regulating the transcription of multiple genes. This implies a potential synergistic effect of eNOS and ZNF580 genes in anti-ischemic therapy. Additionally, we have designed an all-adaptive gene carrier with cascaded bio-responsive functions based on the characteristic bio-signals of ischemic site (including extracellular excessive matrix metalloproteinase-2, endo/lysosomal pH gradient and high cytoplasmic glutathione level). This carrier can sequentially overcome transfection bottlenecks and achieve high transfection. Excitingly, this cascaded bio-responsive delivering strategy has remarkably enhanced blood perfusion, accelerated angiogenesis and alleviated inflammation in critical limb ischemia (CLI) mice, attributing to the combined effect of pro-angiogenic ZNF580 expression and synergistically produced eNOS expression. Thereby, we believe that the co-delivery of eNOS and ZNF580 genes assisted by a cascaded bio-responsive carrier is a powerful strategy to treat CLI.