{"title":"Pancreas-directed AAV8<i>-hSPINK1</i> gene therapy safely and effectively protects against pancreatitis in mice.","authors":"Yuan-Chen Wang, Xiao-Tong Mao, Chang Sun, Ya-Hui Wang, Yi-Zhou Zheng, Si-Huai Xiong, Mu-Yun Liu, Sheng-Han Mao, Qi-Wen Wang, Guo-Xiu Ma, Di Wu, Zhao-Shen Li, Jian-Min Chen, Wen-Bin Zou, Zhuan Liao","doi":"10.1136/gutjnl-2023-330788","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>Currently, there is no cure for chronic pancreatitis (CP). Germline loss-of-function variants in <i>SPINK1</i> (encoding trypsin inhibitor) are common in patients with CP and are associated with acute attacks and progression of the disease. This preclinical study was conducted to explore the potential of adeno-associated virus type 8 (AAV8)-mediated overexpression of human <i>SPINK1</i> (<i>hSPINK1</i>) for pancreatitis therapy in mice.</p><p><strong>Design: </strong>A capsid-optimised AAV8-mediated <i>hSPINK1</i> expression vector (AAV8-<i>hSPINK1</i>) to target the pancreas was constructed. Mice were treated with AAV8-<i>hSPINK1</i> by intraperitoneal injection. Pancreatic transduction efficiency and safety of AAV8-<i>hSPINK1</i> were dynamically evaluated in infected mice. The effectiveness of AAV8-<i>hSPINK1</i> on pancreatitis prevention and treatment was studied in three mouse models (caerulein-induced pancreatitis, pancreatic duct ligation and <i>Spink1</i> c.194+2T>C mouse models).</p><p><strong>Results: </strong>The constructed AAV8-<i>hSPINK1</i> vector specifically and safely targeted the pancreas, had low organ tropism for the heart, lungs, spleen, liver and kidneys and had a high transduction efficiency (the optimal expression dose was 2×10<sup>11</sup> vg/animal). The expression and efficacy of <i>hSPINK1</i> peaked at 4 weeks after injection and remained at significant level for up to at least 8 weeks. In all three mouse models, a single dose of AAV8<i>-hSPINK1</i> before disease onset significantly alleviated the severity of pancreatitis, reduced the progression of fibrosis, decreased the levels of apoptosis and autophagy in the pancreas and accelerated the pancreatitis recovery process.</p><p><strong>Conclusion: </strong>One-time injection of AAV8<i>-hSPINK1</i> safely targets the pancreas with high transduction efficiency and effectively ameliorates pancreatitis phenotypes in mice. This approach is promising for the prevention and treatment of CP.</p>","PeriodicalId":12825,"journal":{"name":"Gut","volume":" ","pages":"1142-1155"},"PeriodicalIF":23.0000,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Gut","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1136/gutjnl-2023-330788","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"GASTROENTEROLOGY & HEPATOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: Currently, there is no cure for chronic pancreatitis (CP). Germline loss-of-function variants in SPINK1 (encoding trypsin inhibitor) are common in patients with CP and are associated with acute attacks and progression of the disease. This preclinical study was conducted to explore the potential of adeno-associated virus type 8 (AAV8)-mediated overexpression of human SPINK1 (hSPINK1) for pancreatitis therapy in mice.
Design: A capsid-optimised AAV8-mediated hSPINK1 expression vector (AAV8-hSPINK1) to target the pancreas was constructed. Mice were treated with AAV8-hSPINK1 by intraperitoneal injection. Pancreatic transduction efficiency and safety of AAV8-hSPINK1 were dynamically evaluated in infected mice. The effectiveness of AAV8-hSPINK1 on pancreatitis prevention and treatment was studied in three mouse models (caerulein-induced pancreatitis, pancreatic duct ligation and Spink1 c.194+2T>C mouse models).
Results: The constructed AAV8-hSPINK1 vector specifically and safely targeted the pancreas, had low organ tropism for the heart, lungs, spleen, liver and kidneys and had a high transduction efficiency (the optimal expression dose was 2×1011 vg/animal). The expression and efficacy of hSPINK1 peaked at 4 weeks after injection and remained at significant level for up to at least 8 weeks. In all three mouse models, a single dose of AAV8-hSPINK1 before disease onset significantly alleviated the severity of pancreatitis, reduced the progression of fibrosis, decreased the levels of apoptosis and autophagy in the pancreas and accelerated the pancreatitis recovery process.
Conclusion: One-time injection of AAV8-hSPINK1 safely targets the pancreas with high transduction efficiency and effectively ameliorates pancreatitis phenotypes in mice. This approach is promising for the prevention and treatment of CP.
期刊介绍:
Gut is a renowned international journal specializing in gastroenterology and hepatology, known for its high-quality clinical research covering the alimentary tract, liver, biliary tree, and pancreas. It offers authoritative and current coverage across all aspects of gastroenterology and hepatology, featuring articles on emerging disease mechanisms and innovative diagnostic and therapeutic approaches authored by leading experts.
As the flagship journal of BMJ's gastroenterology portfolio, Gut is accompanied by two companion journals: Frontline Gastroenterology, focusing on education and practice-oriented papers, and BMJ Open Gastroenterology for open access original research.