Role of GLIS2 in the Development of Cystic Kidney Disease

GLIS2 在囊性肾病发展中的作用

基本信息

批准号：
8730629
负责人：
Massimo Attanasio
金额：
$ 34.58万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-05 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8730629
关键词：
Apoptosis Binding Biological Process Bone Marrow Cells Chimera organism Chronic Kidney Failure Complex Cystic Kidney Diseases Cystic kidney Development Disease Drosophila genus Embryo Epithelial Epithelial Cells Erinaceidae Event Evolution Fibrosis Future GLI Family Protein GLI gene GLI2 gene GLI3 gene Gene Cluster Gene Expression Gene Targeting Genes Genetic High-Throughput Nucleotide Sequencing Human IL1B gene Immune system In Vitro Infiltration Inflammation Inflammatory Inherited Kidney Knock-out Laboratories Life Light Mediating Mediator of activation protein MicroRNAs Modeling Molecular Mus Mutation Natural Immunity Nephronophthisis Orthologous Gene Pathway interactions Phenotype Regulation Role Sequence Homology Signal Transduction Stress Testing Tissues Toll-Like Receptor 2 Toll-like receptors Transcriptional Regulation Tubular formation chromatin immunoprecipitation cytokine effective therapy genome-wide in vivo insight interstitial kidney epithelial cell mouse model mutant overexpression postnatal prevent promoter repaired research study response sensor small hairpin RNA smoothened signaling pathway transcription factor

项目摘要

DESCRIPTION (provided by applicant): Nephronophthisis (NPHP), one of the most frequent genetic causes of chronic renal failure in the first three decades of life, is a group of autosomal recessive diseases characterized by progressive kidney cystic degeneration and fibrosis. Mutations in the gene GLIS2/NPHP7 are the cause of nephronophthisis type 7 both in humans and mice. GLIS2 is a transcription factor with high sequence homology to GLI1, GLI2 and GLI3, the vertebrate orthologs of Drosophila Ci (cubitus interruptus). Like Drosophila Ci, GLI proteins are key molecules in the vertebrate Hedgehog (Hh) signaling, a pathway highly conserved in the evolution and central in the regulation of proliferation, differentiation and repair during embryonc and postnatal life. We have recently tested and confirmed the hypothesis that Glis2 is a repressor of Hh signaling in the postnatal kidney and demonstrated that malfunctioning of this pathway results in kidney cysts and fibrosis, increased inflammatory infiltration and apoptosis, but the molecular events leading to this kidney phenotype are still unexplained. Preliminary studies performed in my laboratory have unexpectedly revealed that loss of Glis2 results in over expression of Toll like receptor 2 (Tlr2) and other components of the pro-inflammatory TLR/NF- ?B pathway in kidney epithelial cells and in the Tlr2-dependent activation of this pathway. We have also found that expression of several miRNAs in kidney epithelial cells is controlled by Glis2, suggesting that microRNAs are effectors of Glis2/Hh signaling in mouse kidneys. We propose to test in vivo the effect of the inhibition of TLR-2/NF-?B signaling on the inflammatory infiltration and fibrosis in Glis2mut/mut mice kidneys by generating Tlr2-/-;Glis2mut/mut and Myd88flox/flox;Glis2mut/mut;KspCre double mutants and establishing Tlr2-/-;Glis2mut/mut bone marrow chimeras. In addition, we propose to systematically identify at genome wide level the canonical and non-canonical Glis2 target genes by chromatin immunoprecipitation-highly parallel sequencing (ChIP-Seq) and high-throughput microRNA sequencing (miRNA-Seq). In light of our most recent results, identifying downstream effectors that are deregulated in Glis2 knockout kidneys has gained even more importance, and is likely to produce further insights in the complex regulatory network that is altered in our model of cystic kidney diseases and fibrosis.

描述（由申请人提供）：肾结核（NPHP）是生命前三十年慢性肾功能衰竭最常见的遗传原因之一，是一组常染色体以进行性肾囊性变和纤维化为特征的隐性疾病。 GLIS2/NPHP7 基因突变是人类和小鼠 7 型肾结核的病因。 GLIS2 是一种与 GLI1、GLI2 和 GLI3（果蝇 Ci（肘节）的脊椎动物直系同源物）具有高度序列同源性的转录因子。与果蝇 Ci 一样，GLI 蛋白是脊椎动物 Hedgehog (Hh) 信号通路中的关键分子，该信号通路在进化中高度保守，并且在胚胎和出生后生命期间的增殖、分化和修复调节中发挥着核心作用。我们最近测试并证实了 Glis2 是出生后肾脏中 Hh 信号传导抑制因子的假设，并证明该通路的故障会导致肾囊肿和纤维化、炎症浸润和细胞凋亡增加，但导致这种肾脏表型的分子事件是仍然无法解释。我的实验室进行的初步研究出乎意料地表明，Glis2 的缺失会导致 Toll 样受体 2 (Tlr2) 以及肾上皮细胞中促炎性 TLR/NF-κB 通路的其他成分以及 Tlr2 依赖性细胞过度表达。该途径的激活。我们还发现肾上皮细胞中几种 miRNA 的表达受 Glis2 控制，表明 microRNA 是小鼠肾脏中 Glis2/Hh 信号传导的效应子。我们建议通过生成 Tlr2-/-;Glis2mut/mut 和 Myd88flox/flox;Glis2mut/ 来体内测试抑制 TLR-2/NF-κB 信号传导对 Glis2mut/mut 小鼠肾脏炎症浸润和纤维化的影响。 mut;KspCre 双突变体并建立 Tlr2-/-;Glis2mut/mut 骨髓嵌合体。此外，我们建议通过染色质免疫沉淀-高度并行测序（ChIP-Seq）和高通量microRNA测序（miRNA-Seq）在全基因组水平上系统地鉴定经典和非经典Glis2靶基因。根据我们最新的结果，识别 Glis2 敲除肾脏中失调的下游效应器变得更加重要，并且可能对我们的囊性肾病和纤维化模型中改变的复杂调控网络产生进一步的见解。