An intronic locus determines SHROOM3-expression and potentiates renal allograft fibrosis

内含子位点决定 SHROOM3 表达并增强肾同种异体移植纤维化

基本信息

批准号：
9749960
负责人：
Barbara T. Murphy
金额：
$ 38.14万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
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：
2015-08-15 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9749960
关键词：
Agonist Alleles Allografting Binding Biopsy Candidate Disease Gene Cells Chronic Chronic Kidney Failure Chronic rejection of renal transplant Collagen Type I Collection Complex Consensus Data Deterioration Development Dominant-Negative Mutation Donor person Drug Targeting End stage renal failure Enhancers Epithelial Cells Etiology European F-actin-binding proteins Fibrosis GTP-Binding Protein alpha Subunits, Gs Gene Expression Genes Genetic Transcription Genomics Genotype HK2 gene Human In Vitro Kidney Kidney Transplantation Link Maps Mediating Messenger RNA Microarray Analysis Modeling Morphology Mus Observational Study Obstruction Outcome Pathogenesis Pathway interactions Phosphorylation Prevention Proteins Protocols documentation Quantitative Trait Loci Regulation Renal function Reporting Research Role Sampling Serum Signal Transduction Single Nucleotide Polymorphism Specific qualifier value TCF7L2 gene Testing Tissues Transcriptional Regulation Transforming Growth Factor beta Transgenic Model Transplant Recipients Transplantation Untranslated RNA Variant allograft rejection base beta catenin cell type cohort diabetic effective therapy in vivo in vivo Model indexing inhibitor/antagonist innovation interstitial kidney allograft kidney fibrosis knock-down mouse model mutant nephrogenesis novel overexpression population based post-transplant prevent promoter prospective protein expression public health relevance risk variant second transplant small hairpin RNA therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Tubulo-interstitial fibrosis (TIF) contributes to the progression of chronic kidney disease (CKD) from varied etiologies, including chronic allograft nephropathy (CAN), to end stage renal failure. However, sensitive predictor of kidney fibrosis and effective antifibrotic therapy for CKD do not exist. Therefore, it is critical for us to identiy markers of and drug targets for TIF. Preliminary data demonstrates that SHROOM3 expression in 3 months renal allografts biopsies was associated with indices of poor allograft function (higher CADI score and decline in eGFR) at 12 months post transplantation, but not at 3 months. Furthermore, theSHROOM3rs17319721 SNP (i.e. A/G or A/A) is associated with a higher SHROOM3 expression at 3 months and a greater propensity to developaΔ CADI (chronic allograft index score) ≥2at 12 months, compared to the G/G allele. In the presence of the risk allele regulation of expression appears to be mediated via a Wnt/βcatenin/ TCF7L2 dependent pathway. Furthermore, expression of SHROOM3 enhanced the profibrotic effects of TGFβ. Based on these preliminary data, the central hypothesize of this proposal is that the SHROOM3 snprs17319721 functions as a cisacting expression quantitative trait locus of SHROOM3 to facilitate TGFβ1 signaling contributes to renal fibrosis leading to progression of CAN and CKD. To test this hypothesis, the following specific aims are proposed: Specific aim 1: To validate that SHROOM3 genotype correlates with expression in a second transplant cohort (1a); To determine the correlation of SHROOM3 genotype and expression with long-term graft outcomes (1b); and To determine SHROOM3 protein expression and localization in the allograft (1c).Specific aim 2: (2a)To determine the transcriptional regulation of SHROOM3 expression by the rs1731972 SNP; (2b) To map the molecular interaction between the TCF7L2/βcatenin complex and the consensus binding sequence encompassing rs17319721; (2c) To investigate the mechanism of SHROOM3facilitated TGFβ signaling in kidney fibrosis. Specific aim 3: To determine the role of SHROOM3 on kidney fibrosis in CAN and CKD murine models in vivo. An inducible and tissues pecific SHROOM3 knockdown model (3a) and an overexpression transgenic model (3b) will be used to examine SHROOM3's function in these models of renal fibrosis.Innovation:SHROOM3is a novel candidate gene for which both a mutant allele and expression predict the decline of renal function and the progression of TIF in renal allografts. This is the first description of a CKD associated SNP that functions as an eQTL. Furthermore, a mechanism for transcriptional regulation via the in tronic SNP is proposed. The current research plan will build upon these initial findings to further delineate transcriptional regulation of SHROOM3, the mechanism by which it contributes the development of fibrosis in vitro and its significance in vivo. The proposed research is of Significance because it will determine the role ofShroom3 in the development and progression of CAN and CKD and its potential as a therapeutic target for the prevention of fibrosis.

描述（由适用提供）：拟南芥纤维化（TIF）有助于慢性肾脏疾病（CKD）从各种病因中的发展，包括慢性同种异体移植肾病（CAN），以结束阶段肾衰竭。但是，不存在肾纤维化和有效的CKD抗纤维化疗法的敏感预测指标。因此，对于我们来说，鉴定TIF的标记和药物靶标对于我们来说至关重要。初步数据表明，在移植后12个月时，Shoom3表达3个月的肾合金活检与合金功能差的指标（EGFR的CADI得分较高和EGFR的较高）相关，但在3个月时却不相关。此外，与g/g相比，theShroom3RS17319721 SNP（即A/G或A/A）与3个月时较高的shroom3表达相关，并且更大的开发PaΔCADI（慢性合金指数得分）≥2AT12个月相比，与G/G等位基因相比。在存在风险等位基因调节的情况下，表达似乎是通过wnt/βcatenin/ tcf7l2依赖途径介导的。此外，Shroom3的表达增强了TGFβ的纤维化作用。基于这些初步数据，该提案的中心假设是Shroom3 SNPRS17319721充当Shroom3的表达表达定量特性，以促进TGFβ1信号传导导致肾纤维化导致CAN和CKD的进展。为了检验这一假设，提出了以下特定目的：特定目的1：验证Shoom3基因型与第二个移植队列中的表达相关（1a）；确定shroom3基因型和表达与长期谷物结局的相关性（1b）；并确定同种异体移植物中的shoom3蛋白表达和定位（1C）。特异性目标2：（2a）确定rs1731972 SNP对shoom3表达的转录调控；（2B）绘制TCF7L2/βCatenin复合物与包含RS17319721的共识结合序列之间的分子相互作用；（2C）研究肾脏纤维化中的shroom3faCiTatedTGFβ信号传导的机制。特定目标3：确定shoom3在cAN中的肾纤维化作用和体内CKD鼠模型的作用。诱导和组织特定的shroom3敲低模型（3A）和过表达的转基因模型（3B）将用于检查这些肾纤维化模型中的shroom3功能。INNOVATION：shroom3IS一个新颖的候选基因，一种突变等位基因和表达的新型候选基因预测肾脏功能的下降和TIF在肾脏同异源性中的下降。这是CKD相关的SNP的第一个描述，该SNP充当eqtl。此外，提出了通过tronic SNP进行转录调控的机制。当前的研究计划将基于这些初步发现，以进一步描述shroom3的转录调节，即chroom3，它促进了体外纤维化的发展及其在体内的重要性。拟议的研究具有重要意义，因为它将确定Shroom3在CAN和CKD的发展和发展中的作用及其作为预防纤维化的治疗靶标的潜力。