Preclinical mouse model of a misfolded PNLIP variant develops chronic pancreatitis

作者：Guoying Zhu, Steven J Wilhelm, Leah G George, Brett M. Cassidy, Sammy Zino, Cliff J. Luke, Mina Hanna, Stephen Stone, Nhung Phan, Neel Matiwala, Samuel Ballentine, Mark E. Lowe, Xunjun Xiao · 发表于：Gut · 年份：2023 · DOI：10.1136/gutjnl-2022-327960 · 被引用次数：24 · 研究领域：Pancreatitis Pathology and Treatment、Diabetes and associated disorders、Inflammatory Myopathies and Dermatomyositis

Objective Increasing evidence implicates mutation-induced protein misfolding and endoplasm reticulum (ER) stress in the pathophysiology of chronic pancreatitis (CP). The paucity of animal models harbouring genetic risk variants has hampered our understanding of how misfolded proteins trigger CP. We previously showed that pancreatic triglyceride lipase (PNLIP) p.T221M, a variant associated with steatorrhoea and possibly CP in humans, misfolds and elicits ER stress in vitro suggesting proteotoxicity as a potential disease mechanism. Our objective was to create a mouse model to determine if PNLIP p.T221M causes CP and to define the mechanism. Design We created a mouse model of Pnlip p.T221M and characterised the structural and biochemical changes in the pancreas aged 1–12 months. We used multiple methods including histochemistry, immunostaining, transmission electron microscopy, biochemical assays, immunoblotting and qPCR. Results We demonstrated the hallmarks of human CP in Pnlip p.T221M homozygous mice including progressive pancreatic atrophy, acinar cell loss, fibrosis, fatty change, immune cell infiltration and reduced exocrine function. Heterozygotes also developed CP although at a slower rate. Immunoblot showed that pancreatic PNLIP T221M misfolded as insoluble aggregates. The level of aggregates in homozygotes declined with age and was much lower in heterozygotes at all ages. The Pnlip p.T221M pancreas had increased ER stress evidenced by dilated ER, increased Hspa5 (BiP)...