O'Brien Kidney Disease Center

总体中心描述（由申请人提供） 这个奥布莱恩肾脏研究中心的主要目标是确定参与上皮钠通道（ENaC）调节的分子机制。这种方法的基本原理是扩大科学框架，从该框架中可以探索盐敏感性高血压的机制。该申请包括三个常规项目和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ENaC活性增加的机制，该机制最近被证明是通过α亚基的羧基末端介导的。内源性激酶活性的作用和四个确定的候选蛋白质的关系，这种调节将被探讨。项目2将研究Nedd 4蛋白家族改变ENaC活性的机制。具体的实验将针对Nedd 4蛋白内的WW结构域与它们的靶标之间的相互作用，PY基序存在于每个ENaC亚基中。项目3将使用基因靶向技术开发一种小鼠，该小鼠可以在他莫昔芬的控制下在集合管主细胞中表达Cre重组酶。这只小鼠将对许多研究人员有潜在的用途，它将被用来开发一种小鼠，这种小鼠在这些细胞中以时间依赖的方式缺乏Nedd 4 -2基因产物的特异性表达。将在预测会产生血压和电解质平衡紊乱的各种条件下检查表型。奥布莱恩中心的第二个目标是培养有前途的年轻科学家，他们将为肾脏疾病的理解和治疗做出贡献。包括两个这样的调查员的试点和可行性研究。一个建议检查一些新发现的蛋白质在肾脏中表达，可能是镁离子通道的具体方面。第二个提出了一种新的方法来治疗肾细胞癌使用腺病毒表达的TRAIL，一种蛋白质引起肿瘤细胞凋亡。奥布莱恩中心将与威尔士博士领导的ENaC/变性蛋白生物学项目和西格蒙德博士领导的高血压分子遗传学SCOR项目进行对接。

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.