O'Brien Kidney Disease Center

整体中心描述(由申请人提供) 这个O‘Brien肾脏研究中心的主要目标是确定参与调节上皮钠通道(ENaC)的分子机制。这种方法的基本原理是拓宽科学框架，从中可以探索盐敏感型高血压的机制。这项申请包括三个常规项目，以及两个试点和可行性研究。常规项目的主题集中在ENaC监管的各个方面。项目1将研究最近被证明是通过阿尔法亚单位的羧基末端介导的ENaC活性增加的机制。将探索内源性激酶活性的作用以及四种已识别的候选蛋白与这一调控的关系。项目2将研究Nedd4蛋白家族改变ENaC活性的机制。具体的实验将针对Nedd4蛋白中的WW结构域与它们的目标之间的相互作用，每个ENaC亚基中都存在PY基序。项目3将利用基因打靶技术开发一种在收集管主细胞中他莫昔芬控制下表达Cre重组酶的小鼠。这种小鼠对许多研究人员都有潜在的用途，它将被用来培育一种在这些细胞中缺乏Nedd4-2基因产物的小鼠，这种小鼠的表达方式与时间有关。表型将在各种条件下进行检测，预计会导致血压和电解质平衡紊乱。这个奥布莱恩中心的第二个目标是促进有前途的年轻科学家的发展，他们将为了解和治疗肾脏疾病做出贡献。两个这样的调查人员的试点和可行性研究也包括在内。一个人建议研究一种新发现的蛋白的某些特定方面，该蛋白在肾脏中表达，可能是一种镁离子通道。第二项研究提出了一种用腺病毒表达TRAIL的方法来治疗肾癌，TRAIL是一种导致肿瘤细胞凋亡的蛋白质。奥布莱恩中心将与威尔什博士领导的ENaC/退化素生物学项目以及西格蒙德博士领导的高血压分子遗传学SCOR项目对接。

DESCRIPTION OF OVERALL CENTER (provided by applicant) The major objective of this O'Brien Kidney Research Center is to identify the molecular mechanisms involved in the regulation of the epithelial Na channel (ENaC). The rationale for this approach is to broaden the scientific framework from which the mechanisms of salt-sensitive hypertension can be explored. This application includes three regular projects, and two pilot and feasibility studies. The themes of the regular projects converge on aspects of ENaC regulation. Project 1 will examine the mechanism for the increase in ENaC activity recently shown to be mediated via the carboxy terminus of the alpha subunit. The role of endogenous kinase activity and the relationship of four identified candidate proteins to this regulation will be explored. Project 2 will examine the mechanisms whereby the Nedd4 family of proteins alters the activity of ENaC. Specific experiments will be directed at the interactions between WW domains within Nedd4 proteins and their targets, the PY motif present in each of the ENaC subunits. Project 3 will use gene targeting to develop a mouse that can express Cre recombinase under the control of tamoxifen in the collecting duct principal cell. This mouse, which will be of potential use to many investigators, will be used to develop a mouse that lacks expression of the Nedd4-2 gene product specifically in those cells in a time dependent fashion. The phenotype will be examined under a variety of conditions predicted to produce derangements in blood pressure and electrolyte balance. A second objective of this O'Brien Center is to foster the development of promising young scientists who will make contributions to the understanding and treatment of kidney diseases. The Pilot and Feasibility Studies of two such investigators are included. One proposes to examine some specific aspects of a newly discovered protein expressed in the kidney that may be a Mg ion channel. The second proposes a novel approach to the treatment of renal cell carcinoma using adenoviral expression of TRAIL, a protein causing apoptosis of tumor cells. The O'Brien Center will interface with the program on the biology of ENaC/degenerins lead by Dr. Welsh and with the SCOR in the Molecular Genetics of Hypertension lead by Dr. Sigmund.