Calcineurin in pancreatitis

描述(申请人提供)：急性胰腺炎是一种痛苦的、危及生命的疾病，目前还没有针对性的治疗方法。我们和其他人已经证明，胰腺腺泡细胞内的病理性钙信号启动了早期胰腺炎反应。此外，我们假设钙依赖的丝氨酸、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来(目的1)研究CN在临床相关的活体胰腺炎实验模型中的作用[1a，胆管内胆酸输注；1b，ERCP后；1c，酒精致敏]，以(目的2)确定腺泡细胞中的CN在胰腺炎中的作用，使用CN抑制剂，腺病毒转染腺病毒构建物，来自CN转基因的细胞，以及(目标3)在体内使用可诱导的，腺泡细胞特异性的CN转基因，要么内源性降低CN的活性，要么删除CN。我们的初步数据支持我们的假设和计划研究的可行性，因为在胆管内胆汁输注的体内模型中，无论是使用CN抑制剂FK506治疗的小鼠，还是CN(CN A2 KOS)基因缺陷的小鼠，都显著降低了胰腺炎的严重程度。我们还表明，在使用化学CN抑制的分离的腺泡细胞或CN缺乏的小鼠细胞中，腺泡内的蛋白酶活性降低。预计这些研究(1)将为了解钙磷酸酶CN在各种形式的胰腺炎中的作用提供基础，(2)将为使用CN抑制剂针对胰腺炎的新的临床试验奠定框架。 与公共卫生相关：急性胰腺炎是一种痛苦的、危及生命的胰腺疾病，我们没有专门的治疗方法。基于胰腺内的钙信号是胰腺炎发生所必需的这一事实，我们的建议试图研究一种重要的钙靶蛋白钙调神经磷酸酶的作用。我们将使用模拟三种常见人类原因的胰腺炎动物模型，即胆结石、酗酒和被称为ERCP的内窥镜手术的并发症。我们希望，我们的研究将导致临床试验，测试已有的钙调神经磷酸酶阻断药物用于治疗胰腺炎。

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. PUBLIC HEALTH RELEVANCE: Acute pancreatitis is a painful and life-threatening disease of the pancreas for which we have no specific treatments. Based on the fact that calcium signals within the pancreas are required for pancreatitis to occur, our proposal seeks to examine the role of an important calcium target protein called calcineurin. We will use animal models of pancreatitis that mimic three common human causes, namely, gallstones, alcohol abuse, and a complication of an endoscopic procedure called an ERCP. We are hopeful that our studies will lead to clinical trials that test the use of already available calcineurin blocking drugs for the treatment of pancreatitis.