Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA.
Department of Gastroenterology, Changhai Hospital, Naval Medical University, Shanghai, China; Shanghai Institute of Pancreatic Diseases, Shanghai, China.
Pancreatology. 2024 Jun;24(4):511-521. doi: 10.1016/j.pan.2024.03.002. Epub 2024 Mar 7.
BACKGROUND & AIMS: Protease-sensitive PNLIP variants were recently associated with chronic pancreatitis (CP) in European populations. The pathological mechanism yet remains elusive. Herein, we performed a comprehensive genetic and functional analysis of PNLIP variants found in a large Chinese cohort, aiming to further unravel the enigmatic association of PNLIP variants with CP.
All coding and flanking intronic regions of the PNLIP gene were analyzed for rare variants by targeted next-generation sequencing in 1082 Chinese CP patients and 1196 controls. All novel missense variants were subject to analysis of secretion, lipase activity, and proteolytic degradation. One variant was further analyzed for its potential to misfold and induce endoplasmic reticulum (ER) stress. p.F300L, the most common PNLIP variant associated with CP, was used as a control.
We identified 12 rare heterozygous PNLIP variants, with 10 being novel. The variant carrier frequency did not differ between the groups. Of them, only the variant p.A433T found in a single patient was considered pathologically relevant. p.A433T exhibited increased susceptibility to proteolytic degradation, which was much milder than p.F300L. Interestingly, both variants exhibited an increased tendency to misfold, leading to intracellular retention as insoluble aggregates, reduced secretion, and elevated ER stress.
Our genetic and functional analysis of PNLIP variants identified in a Chinese CP cohort suggests that the p.A433T variant and the previously identified p.F300L variant are not only protease-sensitive but also may be potentially proteotoxic. Mouse studies of the PNLIP p.F300L and p.A433T variants are needed to clarify their role in CP.
最近,蛋白酶敏感的 PNLIP 变体与欧洲人群的慢性胰腺炎(CP)有关。但病理机制仍不清楚。在此,我们对一个大型中国队列中发现的 PNLIP 变体进行了全面的遗传和功能分析,旨在进一步揭示 PNLIP 变体与 CP 之间神秘的关联。
通过靶向下一代测序,对 1082 例中国 CP 患者和 1196 例对照者的 PNLIP 基因的所有编码和侧翼内含子区域进行罕见变异分析。所有新的错义变异均进行分泌、脂肪酶活性和蛋白水解降解分析。对一种变体进行进一步分析,以研究其是否具有错误折叠和诱导内质网(ER)应激的能力。p.F300L 是与 CP 相关的最常见的 PNLIP 变体,用作对照。
我们发现了 12 种罕见的杂合 PNLIP 变体,其中 10 种是新的。两组之间的变体携带者频率没有差异。其中,只有在单个患者中发现的 p.A433T 变体被认为具有病理学意义。p.A433T 表现出对蛋白水解降解的易感性增加,但其程度远低于 p.F300L。有趣的是,这两种变体都表现出错误折叠的增加趋势,导致细胞内滞留为不溶性聚集体,分泌减少,ER 应激增加。
我们对中国 CP 队列中发现的 PNLIP 变体进行的遗传和功能分析表明,p.A433T 变体和先前鉴定的 p.F300L 变体不仅是蛋白酶敏感的,而且可能是潜在的蛋白毒性的。需要进行 PNLIP p.F300L 和 p.A433T 变体的小鼠研究,以阐明它们在 CP 中的作用。