慢性胰腺炎(CP)是一种慢性不可逆的胰腺纤维炎症性疾病，可导致糖尿病、脂肪泻和癌变等，临床缺乏有效治疗药物。SPINK1基因编码胰酶抑制因子，是抑制胰酶激活的“第一道防线”，其功能缺失型突变可增加胰腺内胰酶自身激活风险。申请者前期发现我国CP主要致病突变为SPINK1 c.194+2T>C（高达42.4%）。既往研究发现，SPINK1突变后或胰酶基因过表达的小鼠模型，均出现CP加重的趋势；而现有目前外源补充人SPINK1蛋白的方法难以长期维持作用。本课题拟构建新型AAV8载体，在人胰腺特异持久表达SPINK1，以抑制胰酶过度激活，从而防治CP的发生与进展。先后分别在体外腺泡细胞和体内CP小鼠模型中验证载体的感染效率和安全性；其次，在CP模型动物中研究AAV8-SPINK1载体对CP发生和进展的影响，为临床CP基因治疗提供临床前应用的证据与理论。

Chronic pancreatitis is defined as a pathological fibro-inflammatory syndrome of the pancreas, clinically featured as progression of endocrine and exocrine insufficiency, and even pancreatic cancer. There is a lack of effective therapeutic drugs in the clinic. The serine protease inhibitor Kazal type 1 gene (SPINK1) encodes the pancreatic secretory trypsin inhibitor, also described as the first line of defense against trypsinogen auto-activation in the pancreas. Loss-of-function variants of SPINK1 has been demonstrated to be associated with the risk of CP. According to our previous studies, SPINK1 c.194+2T>C was identified as the most common pathogenic mutation in Chinese CP patients (up to 42.4%). The mouse models of SPINK1 mutations or PRSS1 (serine protease 1) overexpression could drive the onset and progression of CP. However, human SPINK1 protein supplement therapy couldn’t maintain for a long time in the body. We aim to construct a novel AAV8 vector, which may specifically express SPINK1 in human pancreas to inhibit the over-activation of pancreatic enzyme, thereby preventing the occurrence and progression of CP. First, the transfection efficiency and safety of the vector were verified in in vitro and in vivo. Secondly, the effect of AAV8-SPINK1 vector on the occurrence and progression of CP was studied in CP animal models. This project could lay the theoretical and methodological foundation of gene therapy for CP.