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 propose a “dual genes + all-adaptive carrier” idea. We have innovatively co-delivered eNOS gene and the ZNF₅₈₀ gene encoding its transcription factor to enhance the efficiency of 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 the ischemic site (including extracellular excessive matrix metalloproteinase-2, the 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 delivery strategy remarkably enhanced blood perfusion, accelerated angiogenesis and alleviated inflammation in critical limb ischemia (CLI) mice, which was attributed to the combined effects 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.

中文翻译：

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eNOS基因和ZNF580基因的级联生物反应性传递，通过促血管生成和抗炎作用共同治疗后肢缺血

基因疗法是解决缺血和炎症的双重难题，是一种治疗缺血性疾病的有前途的策略。但是，其发展仍然受到无效基因转染的限制。因此，我们提出了“双基因+全适应载体”的构想。我们创新地共同提供了eNOS基因和编码其转录因子的ZNF ₅₈₀基因，以增强eNOS表达的效率。过表达的ZNF ₅₈₀蛋白通过调节多个基因的转录显着促进血管生成。这意味着eNOS和ZNF ₅₈₀的潜在协同作用基因在抗缺血治疗中的作用。此外，我们基于缺血部位的特征生物信号（包括细胞外过量基质金属蛋白酶-2，内/溶酶体pH梯度和高细胞质谷胱甘肽水平）设计了具有级联生物响应功能的全适应基因载体。该载体可以顺序克服转染瓶颈并实现高转染。令人兴奋的是，这种级联的生物反应性递送策略显着增强了关键肢体缺血（CLI）小鼠的血液灌注，加速了血管生成并减轻了炎症，这归因于促血管生成的ZNF ₅₈₀表达和协同产生的eNOS表达。因此，我们认为eNOS和ZNF _580可以共同交付 由级联的生物响应性载体辅助的基因是治疗CLI的有效策略。