Calcineurin in pancreatitis

描述（由申请人提供）：急性胰腺炎是一种痛苦的、危及生命的疾病，目前尚无靶向治疗方法。我们和其他人已经表明胰腺腺泡细胞内的病理性钙信号启动早期胰腺炎反应。此外，我们假设钙依赖性丝氨酸、苏氨酸磷酸酶钙调神经磷酸酶（CN）是这种病理性钙信号的新靶点。因此，我们建议使用遗传学和药理学策略（目的1）检查CN在体内胰腺炎临床相关实验模型中的作用[1a，导管内胆汁酸输注; 1b，ERCP后; 1c，酒精致敏的]，以（目的2）使用CN抑制剂，CN构建体的腺病毒转染，和来自CN转基因的细胞，以及（目的3）使用内源性降低CN活性或缺失CN的可诱导的腺泡细胞特异性CN转基因进行体内研究。我们的初步数据支持我们的假设和计划研究的可行性，表明在导管内胆汁输注的体内模型中，CN抑制剂FK 506或CN基因缺陷（CN A2科斯）的小鼠显著降低了胰腺炎的严重程度。我们还表明，在分离的腺泡细胞中使用化学CN抑制或CN缺陷小鼠的细胞，腺泡内蛋白酶激活减少。预计这些研究（1）将为理解钙磷酸酶CN在各种形式胰腺炎中的作用提供基础，（2）将为使用CN抑制剂靶向胰腺炎的新型临床试验奠定框架。

DESCRIPTION (provided by applicant): Acute pancreatitis is a painful, life-threatening disorder for which there are no targeted therapies. We and others have shown that pathologic calcium signals within the pancreatic acinar cell initiate early pancreatitis responses. Further, we have hypothesized that the calcium-dependent serine, threonine phosphatase calcineurin (CN) is a novel target of this pathological calcium signal. Thus, we propose to use genetic and pharmacologic strategies to (Aim 1) examine the role of CN in clinically relevant experimental models of pancreatitis in vivo [1a, intra-ductal bile acid infusion; 1b, post-ERCP; 1c, alcohol sensitized], to (Aim 2) determine the contribution of acinar cell CN to pancreatitis in isolated acinar cells using CN inhibitors, adenoviral transfection of CN constructs, and cells from CN transgenics, as well as (Aim 3) in vivo using inducible, acinar cell specific CN transgenics that either endogenously reduce CN activity or delete CN. Our preliminary data support our hypothesis and the feasibility of the planned studies by showing that mice either treated with the CN inhibitor FK506 or genetically deficient in CN (CN A2 KOs) have markedly reduced pancreatitis severity in an in vivo model of intra-ductal bile infusion. We also show that in isolated acinar cells using chemical CN inhibition or cells from CN deficient mice that intra-acinar protease activation is reduced. It is anticipated that these studies (1) will provide a basis for understanding the role of the calcium phosphatase CN in various forms of pancreatitis and (2) will lay the framework for novel clinical trials that target pancreatitis using CN inhibitors.