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）：在过去的赠款周期中，使用多种大鼠模型，我们证明了Rab38负责定量性状特质基因座（QTL），名为RF-2，并确定并确认了SORCS1为QTL造成了名为RF-1的QTL，并确定了shroom3在glomerulul glomelular glomelulul gromelulul nullulululululululular enlullulululular nullulululululular enthroom enperealulul gromerulul enerulull enteralulul enterlulul gromerulul enterclible。我们已经朝克隆的位置取得了长足的进步，两个QTL，RF-3和RF-4。与慢性肾脏疾病遗传学（CKDGEN）财团合作，我们发现Sorcs1和Shroom3与人类的肾脏疾病显着相关，表明这项工作具有临床意义。 我们的长期目标是定义终阶段肾脏疾病（ESRD）的遗传结构。我们的数据表明，蛋白尿中需要肾小管近端小管中的肾小球通透性和蛋白质运输的缺陷，这是我们的Centrl假设。提出的具体目的是1）测试shroom3的人类序列变体的集合，预测功能失调； 2）确定参与肾小球通透性的QTL RF-3（间隔大小1.81 MB）和QTL RF-4（间隔尺寸0.9 MB）的基因/基因组元素，认为这启动了蛋白尿； 3）检验的中心假设是，近端小管中肾小球通透性和蛋白质运输的功能障碍都参与了蛋白尿的发展。测试这个中心 假设需要全动物的使用，不能在人类中进行。我们针对该提案的遗传/基因组基础设施包括：父母菌株的完整基因组序列，包括生物信息学分析，“狭窄”的先天性菌株，三个确定的病变基因，以及详细的生理特征以及为发现重要的新基因的理想平台和测试这种新的中央假说创造理想的生理特征。我们将研究HEK293细胞，斑马鱼和人源化的FHH大鼠模型的HEK293细胞中的人类变体。具体而言，AIM 1测试了针对肾脏疾病的人类变体的集合，并证明了如何使用模型系统来跟进许多全基因组关联研究（GWAS）提名的基因。我们与CKDGEN的合作使我们能够测试针对RF-3和RF-4确定的基因是否有助于人类疾病。因此，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了人类在Shroom3中的变异，新基因的鉴定以及新的假设将为这一疾病过程提供新的见解，并可以确定新的目标，以升高和有效的治疗方法，以使肾脏疾病的未来治疗。具体目的1-3有望揭示有关shroom3功能的知识并识别两个新基因，它们将链接到已知途径或揭示新基因。我们还将确定是否涉及两次命中（肾小球中的一个和一个近端小管中）。该提案的每个目标都将为定义蛋白尿的遗传结构提供基本的进步，蛋白尿通常是ESRD的前体。