Physiological Genomics of Hypertensive Renal Disease

描述(由申请人提供)：ESRD是美国的一个主要健康问题，发病率不断上升。高血压和糖尿病占ESRD的67%以上。这些关联尚不清楚，但许多人认为，肾功能衰竭存在一条最终的共同途径，不依赖诱导剂或疾病。并不是每个患有高血压或糖尿病的人都会患上终末期肾病，我们证明了肾衰竭基因可以独立于高血压。我们有一个综合的研究计划，利用遗传学、基因组学、转基因技术来研究高血压、糖尿病和肾脏疾病易感基因的相互作用。这项建议的目标是确定与高血压相关的ESRD易感性相关的基因，并确定涉及的生理途径。我们提出三个具体目标： 1.验证Rab38为RF-2基因。我们最近报道了Rab38是RF-2基因。我们推测Rab38基因具有多效性，可能与FHH大鼠蛋白尿的发生有关。我们将通过a)研究近端小管原代培养中白蛋白的摄取和b)转基因“拯救”来验证这一假设。 2.完成了RF-1基因和RF-4基因的定位克隆和验证。在FHH中，Rf-1 QTL是ESRD的显性座位。重要的是，该基因座与RF-4有很强的相互作用。我们已经将RF-1间隔减少到2.5 cM，将RF-4间隔减少到0.8 cM，并且BN大鼠的整个区域已经测序到接近完成的水平。我们将使用亚基因、生理研究和序列比较的方法对每个基因座进行调查，以确定两个基因座中的致病基因(S)。因果基因将在FHH基因组背景下通过大鼠转基因拯救来验证。 3.启动RF-3的定位克隆。我们已经证明了我们有能力通过位置克隆基因。我们发现，在小鼠和人类中，RF-3也位于与肾脏疾病相关的区域，就像RF-1、-2和-4一样。我们将使用现有的双(RF-1+3)同基因株在RF-3区间产生重叠的同源基因。我们相信，当我们完成这项资助的期限时，我们将要么找到这个基因，要么将间隔缩小到少数几个基因。

DESCRIPTION (provided by applicant): ESRD is a major health problem in the United States, with increasing incidence. Hypertension and diabetes account for more than 67% of ESRD. These associations are not understood, but many believe there is a final common pathway for renal failure independent of inducing agent or disease. Not everyone with hypertension or diabetes develops ESRD, and we demonstrated that renal failure genes could be independent of hypertension. We have a comprehensive research program utilizing genetics, genomics, transgenics to study the interactions of hypertension, diabetes and renal disease susceptibility genes. The goal of this proposal is to identify genes responsible for susceptibility to hypertension-related ESRD, and to identify the physiological pathways involved. We propose three specific aims: 1. Validate that Rab38 is the Rf-2 gene. We recently reported Rab38 to be the Rf-2 gene. We hypothesize that the Rab38 gene has pleiotropic effects and contributes to the development of proteinuria in FHH rats. We will test this hypothesis by a) studying albumin uptake in primary cultures of proximal tubule and b) with transgenic "rescue". 2. Complete the positional cloning and validation of the Rf-1 gene and Rf-4 gene. The Rf-1 QTL is the dominant locus for ESRD in the FHH. Importantly, this locus has a strong interaction with Rf-4. We have reduced both the Rf-1 interval to 2.5 cM and the Rf-4 interval to 0.8 cM, and the entire region has been sequenced in BN rats to a nearly finished level. We will investigate each locus using sub-congenics, physiological studies and sequence comparison to identify the causal gene(s) in both loci. Causal genes will be validated by rat transgenic rescue in the FHH genome background. 3. Initiate the positional cloning of Rf-3. We have demonstrated our ability to clone genes by position. We show that Rf-3 is also in a region linked to renal disease in mouse and human as is the case for Rf-1, -2, and -4. We will use the existing double (Rf-1+3) congenic strain to generate the overlapping congenics in the Rf-3 interval. We are confident that we will either find the gene or narrow down the interval to a handful of genes when we complete the term of this grant.