糖尿病患者勃起功能障碍（ED）发病率高，药物疗效欠佳，探索新的治疗方法和机制是目前研究的主要方向，基因治疗是研究热点，其中靶点和载体选择尤为重要。NO-cGMP通路的PDE5是目前治疗ED的首选靶点。申请者先前研究表明JAK2通路在糖尿病ED中起重要作用，该通路激活可导致平滑肌细胞损伤、舒张功能下降而导致ED。外泌体是一类非病毒非细胞物质的生物小囊泡，具有免疫原性低、可向靶细胞定向移动的特性，被认为是一种理想的基因载体。本项目拟通过构建搭载CRISPRi系统的平滑肌外泌体，将其注射入阴茎海绵体内，靶向局灶敲减经典PDE5基因或/和申请者所研究的JAK2基因，增加平滑肌cGMP含量，促进平滑肌损伤修复，提高ED治疗效率；同时在细胞和分子水平探讨外泌体介导CRISPRi调控PDE5、JAK2基因增强糖尿病ED大鼠勃起功能可能的作用机制，这将为外泌体运载的基因治疗糖尿病ED提供科学依据。

Erecrtile dysfunction is prevalent in diabetic patients, who have a poor response to PDE5 inhibitors. To explore novel modalities and mechanisms is the future for ED treatment. Current researches mainly focus on gene therapy, especially on therapeutic targets and proper vehicles. PDE5 in NO-cGMP pathway is the prior target for ED treatment. Our previous study has proven that JAK2 pathway plays a vital role in diabetic erectile dysfunction. The activation of JAK2 pathway could lead ED via injuring cavernous smooth muscle and dampening the relaxation of smooth muscle. Exosomes are non-viral non-cell biovesicles, possessing low immunogenicity and the ability to migrate to targeted cells, which is an ideal carrier for gene therapy. Current study is to construct cavernous smooth muscle cell-derived exosomes equipped with CRISPRi system and to focally knock down targeted classical PDE5 or / and JAK2 genes studied by applicants. Aims to increase cGMP concentration and promote the regeneration of smooth muscle cells in the penis. We will explore the possible mechanisms related to this modality. The aforementioned study will provide evidence for the potential exosome based gene therapy for diabetic erectile dysfunction.