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

基本信息

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
项目类别：
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

项目摘要

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，使用多种大鼠模型，我们已经证明 Rab38 负责名为 Rf-2 的数量性状基因座（QTL），并鉴定并验证了 Sorcs1 负责 QTL命名为 Rf-1，并确定 Shroom3 会导致肾小球通透性变化。我们在通过另外两个 QTL Rf-3 和 Rf-4 进行克隆方面取得了长足的进步。我们与慢性肾脏病遗传学 (CKDGen) 联盟合作，发现 SORCS1 和 SHROOM3 与人类肾脏疾病显着相关，证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，蛋白尿需要肾小球通透性和近曲小管中蛋白质运输的缺陷，这也是我们现在的中心假设。拟议的具体目标是 1) 测试预计功能失调的 SHROOM3 人类序列变体集合； 2) 鉴定参与肾小球通透性（被认为引发蛋白尿）的 QTL Rf-3（间隔大小 1.81 Mb）和 QTL Rf-4（间隔大小 0.9 Mb）背后的基因/基因组元件； 3）检验中心假设，即肾小球通透性和近端肾小管蛋白质运输功能障碍共同参与蛋白尿的发生。测试这个中心假设需要使用整个动物，而不能在人类身上进行。我们针对该提案的遗传/基因组基础设施包括：亲本菌株的完整基因组序列、NextGen 测序工具（包括生物信息学分析）、“窄”同源菌株、三个已识别的致病基因以及详细的生理特征，为发现重要的新基因创建了一个理想的平台并检验这个新的中心假设。我们将研究 HEK293 细胞、斑马鱼和人源化致敏 FHH 大鼠 CKD 模型中 SHROOM3 的人类变体。具体来说，Aim 1 测试了一系列人类肾脏疾病变异，并展示了如何使用模型系统来追踪许多全基因组关联研究 (GWAS) 指定的基因。我们与 CKDGen 的合作使我们能够测试 Rf-3 和 Rf-4 基因是否会导致人类疾病；从而提供有关人类疾病过程的知识。拟议研究的基本原理是测试 SHROOM3 的人类变异、识别新基因和新假设，该假设将为这一疾病过程提供新见解，并为未来治疗发病率上升和有限的肾脏疾病确定新目标。有效的治疗。具体目标 1-3 预计将揭示有关突变的 SHROOM3 如何发挥作用的知识，并识别两个新基因，这些基因要么与已知的途径相关，要么揭示新的途径。我们还将确定是否涉及两次打击（一次在肾小球，一次在近端小管）。该提案的每个目标都将为定义蛋白尿的遗传结构提供根本性的进展，蛋白尿通常是终末期肾病的前兆。