Evaluation of human variants in disease models for end stage renal disease

终末期肾病疾病模型中人类变异的评估

• 批准号: 9116554
• 负责人: HOWARD J JACOB
• 金额: $ 13.24万
• 依托单位: HUDSON-ALPHA INSTITUTE FOR BIOTECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9116554
• 关键词: Affect; Alleles; Animal Model; Animals; Architecture; Bioinformatics; Biological Models; Cells; Chronic Kidney Failure; Cloning; Collaborations; Collection; Congenic Strain; Data; Defect; Development; Diabetes Mellitus; Disease; Disease model; Elements; Embryo; End stage renal failure; Epidemiologic Studies; Evaluation; Focal glomerulosclerosis; Functional disorder; Future; Gene Expression; Genes; Genetic; Genetic Predisposition to Disease; Genetic screening method; Genetic study; Genomics; Goals; Grant; Health; Human; Hypertension; Incidence; Kidney; Kidney Diseases; Knock-out; Knowledge; Letters; Link; Malignant neoplasm of prostate; Modeling; Mutate; Names; Pathway interactions; Patients; Permeability; Physiological; Play; Positioning Attribute; Prevalence; Process; Proteins; Proteinuria; Quantitative Trait Loci; Rattus; Reporting; Research; Research Infrastructure; Role; Survival Rate; Testing; Variant; Work; Zebrafish; clinically relevant; cohort; congenic; design; effective therapy; embryonic stem cell; follow-up; gene discovery; genome wide association study; glomerular function; human data; human disease; hypertension treatment; improved; in vivo Model; insight; malignant breast neoplasm; next generation sequencing; novel; podocyte; protein transport; public health relevance; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): During the past grant cycle, using a variety of rat models, we have proven that Rab38 is responsible for the quantitative trait locus (QTL) named Rf-2, identified and validated that Sorcs1 is responsible for the QTL named Rf-1, and have identified that Shroom3 causes changes in glomerular permeability. We have made considerable strides towards cloning by position two other QTL, Rf-3 and Rf-4. In collaboration with the Chronic Kidney Disease Genetics (CKDGen) Consortium, we found that SORCS1 and SHROOM3 are significantly associated with renal disease in humans, demonstrating that this work has clinical relevance. Our long-term goal is to define the genetic architecture of end stage renal disease (ESRD). Our data suggests that a defect in both the glomerular permeability and protein trafficking in the proximal tubules is required for proteinuria, which is now our centrl hypothesis. The proposed Specific Aims are to 1) test a collection of human sequence variants of SHROOM3 predicted to be dysfunctional; 2) identify the genes/genomic elements underlying the QTL Rf-3 (interval size 1.81 Mb) and QTL Rf-4 (interval size 0.9 Mb) participating in glomerular permeability, which is thought to initiate proteinuria; and 3) test the central hypothesis that dysfunction in both glomerular permeability and protein trafficking in the proximal tubules are participating together in the development of proteinuria. Testing this central hypothesis requires the use of whole animals and cannot be done in human. Our genetic/genomic infrastructure for this proposal includes: complete genomic sequence of parental strains, NextGen sequencing tools including bioinformatic analysis, "narrow" congenic strains, three identified causative genes, as well as detailed physiological characterization creating an ideal platform for discovering important new genes and testing this new central hypothesis. We will study human variants of SHROOM3 in HEK293 cells, zebrafish and humanized sensitized FHH rat model for CKD. Specifically, Aim 1 tests a collection of human variants for renal disease and demonstrates how model systems can be used to follow-up genes nominated by many genome-wide association studies (GWAS). Our collaboration with the CKDGen enables us to test if the genes identified for Rf-3 and Rf-4 contribute to human disease; thereby, providing knowledge about the disease process in humans. The rationale for the proposed research is the test of human variation in SHROOM3, identification of new genes, and a new hypothesis which will provide new insights into this disease process and could identify new targets for future treatment of renal disease with a rising incidence and limited effective treatments. Specific aims 1-3 are expected to reveal the knowledge about how mutated SHROOM3 functions and identify two new genes, which will either link to known pathways or unmask new ones. We will also determine if two hits (one in the glomeruli and one in proximal tubules) are involved. Each aim of this proposal will provide fundamental advances toward defining the genetic architecture of proteinuria which is often the precursor of ESRD.

描述（由申请人提供）：在过去的资助周期中，我们使用多种大鼠模型证明了Rab38负责数量性状位点（QTL） Rf-2，鉴定并验证了Sorcs1负责数量性状位点（QTL） Rf-1，并鉴定了Shroom3引起肾小球通透性的变化。我们已经通过另外两个QTL， Rf-3和Rf-4，在克隆方面取得了相当大的进展。在与慢性肾脏疾病遗传学（CKDGen）联盟的合作中，我们发现SORCS1和SHROOM3与人类肾脏疾病显著相关，表明这项工作具有临床意义。我们的长期目标是确定终末期肾病（ESRD）的遗传结构。我们的数据表明，蛋白尿需要肾小球渗透性和近端小管中蛋白质运输的缺陷，这是我们现在的中心假设。提出的具体目标是：1)测试预测功能失调的SHROOM3人类序列变体的集合；2)鉴定参与肾小球通透性的QTL Rf-3（间隔大小1.81 Mb）和QTL Rf-4（间隔大小0.9 Mb）的基因/基因组元件，肾小球通透性被认为是蛋白尿的起始；3)验证肾小球通透性和近端小管蛋白质运输功能障碍共同参与蛋白尿发生的中心假设。测试这个中心