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Biochemical and Functional Studies of Sall1 in Kidney Development and TBS

Sall1 在肾脏发育和 TBS 中的生化和功能研究

基本信息

批准号：
7939666
负责人：
MICHAEL I RAUCHMAN
金额：
$ 31.1万
依托单位：
SAINT LOUIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-08 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7939666
关键词：
Affect Alleles Binding Biochemical Biochemical Genetics Biological Assay Breeding Cell Line Cell physiology Cells Chromatin Clinical Complex Congenital Abnormality Data Deacetylase Defect Dependence Development Disease Electroporation Embryo Etiology Event Exhibits Gel Chromatography Gene Expression Gene Targeting Genes Genetic Goals Heterozygote Human Immunohistochemistry Immunoprecipitation In Situ Hybridization Inherited Investigation Kidney Knock-in Mouse Knockout Mice Lead Maintenance Mediating Mesenchyme Metanephric Diverticulum Metanephric structure Microinjections Middle Lobe Syndrome Modeling Molecular Molecular Genetics Morphogenesis Multiprotein Complexes Mus Mutant Strains Mice Mutation Nephrons Nucleosomes Organ Organ Culture Techniques Organogenesis Pathogenesis Pathway interactions Patients Phenotype Process Promoter Regions Proteins Publishing RNA Interference Recruitment Activity Regulation Reporter Repression Reverse Transcriptase Polymerase Chain Reaction Role Signal Transduction Staging Stromal Cells Syndrome Testing Time Tissues Transgenic Mice Tumor stage Ureter Urinary tract Zinc Fingers base blastomere structure cell type chromatin immunoprecipitation gain of function in vivo insight loss of function mRNA Expression mouse model mutant nephrogenesis novel novel therapeutic intervention progenitor programs protein complex public health relevance recombinase transcription factor

项目摘要

DESCRIPTION (provided by applicant): Organogenesis requires the precise temporal and spatial control of gene expression to direct developmental programs that regulate a variety of cellular processes. Sall genes encode for multi-zinc finger transcription factors that are critical conserved regulators of organ development. Mutations in SALL1 cause the Townes-Brocks syndrome (TBS), an autosomal dominant disorder with developmental defects affecting multiple organs. Anomalies of the kidney are prominent in this condition. We created a mouse model of TBS and showed that this syndrome results from dominant effects of a Sall1N truncated mutant protein. These mice display renal hypoplasia with reduced branching of the ureter, similar to that seen in TBS patients. In Sall1 homozygous mutant mice the ureter fails to properly initiate branching resulting in renal agenesis. While these studies indicate that Sall1 is essential for kidney development, the molecular and developmental mechanisms of its function are not well understood. To delineate the function of Sall1 in the kidney it is important to identify Sall1 target genes and determine how these downstream pathways regulate branching morphogenesis of the ureteric bud, maintenance of metaneprhic progenitors and nephron formation. We discovered a novel repression motif in Sall1 (SRM) that is necessary and sufficient to recruit the nucleosome remodeling and deacetylase (NuRD) complex to mediate repression of native Sall1 target genes. Our current model is that regulation of an important subset of Sall1 target genes in developing kidney depends on NuRD recruitment. Based on our preliminary data it is likely that Sall1 acts at multiple steps during nephrogenesis. However, the early arrest of metanephric development exhibited by existing Sall1 alleles has limited investigation of these processes. Since Sall1 is expressed in both the cap mesenchyme (metanephric progenitors) and stromal cells, defining its function in these distinct cellular compartments and at different developmental stages is an important question in the field. These ideas will be tested in three specific aims: (1) Identification and functional analysis of direct Sall1 target genes in metanephric mesenchyme (2) Investigate the role of Sall1 in cap mesenchyme and stromal cells in developing kidney, and (3) Determine the composition of Sall1 native protein complexes in embryonic kidney. These aims will increase our understanding of Sall1 function in nephrogenesis and the etiology of TBS. PUBLIC HEALTH RELEVANCE: Congenital anomalies affecting the kidney and urinary tract are among the most common serious birth defects. While there is significant clinical and increasing genetic data on these disorders, the disease mechanisms are not well understood. The proposed studies will elucidate the molecular pathogenesis of these inherited organ defects and this in turn may lead to novel therapeutic approaches.

描述（由申请人提供）：器官发生需要精确的时间和空间控制基因表达，以指导调节各种细胞过程的发育程序。萨尔基因编码多锌指转录因子，这些因子是器官发育的关键保守调节因子。SALL 1突变导致Towns-Brocks综合征（TBS），这是一种常染色体显性遗传疾病，具有影响多个器官的发育缺陷。在这种情况下，肾脏的异常是突出的。我们建立了一个TBS小鼠模型，并表明这种综合征是由Sall 1 N截短突变蛋白的显性效应引起的。这些小鼠表现出肾发育不全，输尿管分支减少，与TBS患者相似。在Sall 1纯合子突变小鼠中，输尿管不能正确启动分支，导致肾发育不全。虽然这些研究表明Sall 1对肾脏发育至关重要，但其功能的分子和发育机制尚不清楚。为了阐明Sall 1在肾脏中的功能，重要的是要确定Sall 1的靶基因，并确定这些下游途径如何调节输尿管芽的分支形态发生，后肾祖细胞的维持和肾单位的形成。我们在Sall 1（SRM）中发现了一个新的阻遏基序，该基序对于招募核小体重塑和脱乙酰酶（NuRD）复合物来介导天然Sall 1靶基因的阻遏是必要且足够的。我们目前的模型是，在发育中的肾脏中Sall 1靶基因的重要子集的调节取决于NuRD募集。根据我们的初步数据，Sall 1可能在肾发生过程中的多个步骤中起作用。然而，现有的Sall 1等位基因表现出的后肾发育的早期停滞限制了对这些过程的研究。由于Sall 1在帽间充质（后肾祖细胞）和基质细胞中表达，因此定义其在这些不同细胞隔室和不同发育阶段的功能是该领域的一个重要问题。本研究将从以下三个方面对Sall 1基因在后肾间充质中的作用进行研究：（1）Sall 1基因在后肾间充质中的直接靶基因的鉴定和功能分析;（2）Sall 1基因在肾帽间充质细胞和间质细胞中的作用研究;这些目标将增加我们对Sall 1在肾发生和TBS病因学中的功能的理解。公共卫生相关性：影响肾脏和泌尿道的先天性异常是最常见的严重出生缺陷之一。虽然有关于这些疾病的重要临床和越来越多的遗传数据，但疾病机制尚未得到很好的理解。拟议的研究将阐明这些遗传性器官缺陷的分子发病机制，这反过来可能导致新的治疗方法。