Trypsin-dependent mechanisms in pancreatitis

胰腺炎的胰蛋白酶依赖性机制

基本信息

批准号：
9916956
负责人：
Miklos Sahin-Toth
金额：
$ 48.78万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9916956
关键词：
Acinar Cell Acute Adipose tissue Advanced Development Applications Grants Atrophic Biochemical Biological Cations Cell Death Cells Chronic Data Disease Disease Progression Disease model Enzymes Exhibits Fibrosis Future Genes Genetic Predisposition to Disease Genetic Risk Genetically Engineered Mouse Grant Human Human Genetics In Vitro Infiltration Inflammatory Inflammatory Response Injections Injury Mediating Modeling Molecular Mouse Strains Mus Mutation Onset of illness Orthologous Gene Pancreas Pancreatic Diseases Pancreatic Injury Pancreatitis Pathogenicity Pathologic Pathology Pathway interactions Peptides Phenotype Predisposition Preventive Intervention Protein Isoforms Protein secretory trypsin inhibitor Protocols documentation Rattus Research Risk Factors Role SPINK1 gene Serine Protease Serine Proteinase Inhibitors Testing Therapeutic Tissues Transgenic Organisms Trypsin Trypsin Inhibitors Trypsinogen acute pancreatitis base chronic pancreatitis chymotrypsin C design endoplasmic reticulum stress genetic association genetic risk factor hereditary pancreatitis in vivo in vivo Model inhibitor/antagonist loss of function mutation mutant mouse model non-alcoholic novel novel therapeutic intervention overexpression pre-clinical programs response risk variant

项目摘要

ABSTRACT The main objective of this grant is to use genetically engineered mouse models to determine the mechanisms by which trypsinogen (PRSS1) mutations in humans cause hereditary pancreatitis. The majority of non-alcoholic cases of chronic pancreatitis develop on the basis of genetic susceptibility, driven by mutations in risk genes that encode digestive enzymes or their inhibitor, such as cationic trypsinogen (protease serine 1, PRSS1), chymotrypsin C (CTRC) or the pancreatic secretory trypsin inhibitor (serine protease inhibitor Kazal type 1, SPINK1). Our previous studies defined a trypsin- dependent pathological pathway associated with mutations in these risk genes that promote intra- pancreatic trypsinogen autoactivation and result in increased ectopic trypsin activity. In the present proposal, we will validate this model in vivo. To this end, we developed novel mouse lines T7 D23A and T7 K24R that carry mutations in the activation peptide of the native mouse cationic trypsinogen (isoform T7). In vitro the D23A mutation causes a dramatic 50-fold increase in trypsinogen autoactivation, while mutation K24R increases autoactivation by 4-fold. Thus, the models can provide information on how different trypsin levels determine pancreatitis responses and pathology. We hypothesize that mice with trypsinogen mutations that stimulate autoactivation will develop spontaneous pancreatitis or exhibit heightened pancreatitis responses when challenged with hyperstimulation insults. In our specific aims we will study the spontaneous pancreatitis in the T7 D23A mouse; evaluate the increased sensitivity to secretagogue induced pancreatitis in the T7 K24R mouse and investigate the protective role of trypsin inhibition against pancreatitis by altering Spink3 (ortholog of human SPINK1) expression levels in these models. Successful completion of these aims will firmly establish that increased trypsinogen autoactivation leading to elevated intra-pancreatic trypsin activity is a relevant disease-mechanism in pancreatitis and should be the focus of future therapeutic strategies.

抽象的这笔赠款的主要目标是使用基因工程小鼠模型来确定人类胰蛋白酶原（PRSS1）突变导致遗传性胰腺炎的机制。这大多数非酒精性慢性胰腺炎病例是在遗传易感性的基础上发展的，由编码消化酶或其抑制剂（例如阳离子）的风险基因突变驱动胰蛋白酶原（丝氨酸蛋白酶 1，PRSS1）、胰凝乳蛋白酶 C (CTRC) 或胰腺分泌胰蛋白酶抑制剂（丝氨酸蛋白酶抑制剂 Kazal 1 型，SPINK1）。我们之前的研究定义了胰蛋白酶- 与这些风险基因突变相关的依赖病理途径，促进内部胰腺胰蛋白酶原自动激活并导致异位胰蛋白酶活性增加。在现在建议，我们将在体内验证该模型。为此，我们开发了新型小鼠品系 T7 D23A 和 T7 K24R 携带天然小鼠阳离子胰蛋白酶原（异构体）激活肽的突变 T7）。在体外，D23A 突变导致胰蛋白酶原自动激活急剧增加 50 倍，而 K24R 突变使自激活增加 4 倍。因此，模型可以提供有关如何不同的胰蛋白酶水平决定胰腺炎的反应和病理。我们假设小鼠刺激自身激活的胰蛋白酶原突变会发展为自发性胰腺炎或表现出当受到过度刺激侮辱时，胰腺炎反应会加剧。在我们的具体目标中我们将研究 T7 D23A 小鼠的自发性胰腺炎；评估增加的敏感性 T7 K24R 小鼠促分泌剂诱导的胰腺炎并研究胰蛋白酶的保护作用通过改变 Spink3（人 SPINK1 的直系同源物）表达水平来抑制胰腺炎模型。成功完成这些目标将牢固地确定胰蛋白酶原的增加导致胰腺内胰蛋白酶活性升高的自身激活是相关的疾病机制胰腺炎应成为未来治疗策略的重点。