Deregulation of Long Noncoding RNA in the Pathogenesis of Autosomal Dominant Polycystic Kidney Disease

常染色体显性多囊肾病发病机制中长非编码 RNA 的失调

基本信息

批准号：
10378018
负责人：
Karam Aboudehen
金额：
$ 10.85万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-15 至 2022-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10378018
关键词：
70-kDa Ribosomal Protein S6 Kinases Affect Age Applications Grants Autophagocytosis Autosomal Dominant Polycystic Kidney Basic Science Binding Bioinformatics Biological Assay Biology Biometry Cell Proliferation Cells Cellular Metabolic Process Clustered Regularly Interspaced Short Palindromic Repeats Code Complex Computing Methodologies Cyst Cystic Kidney Diseases Cystic kidney Data Analyses Development Dialysis procedure Disease Disease Progression Down-Regulation EIF4EBP1 gene End stage renal failure Epigenetic Process Epithelial Exhibits FDA approved FRAP1 gene Functional disorder Future Genes Genetic Genetic Crosses Genetic Diseases Genomics Goals Growth High-Throughput Nucleotide Sequencing Human Immunoprecipitation In Vitro Institution K-Series Research Career Programs Kidney Kidney Transplantation Knock-out Knockout Mice Laboratories Leadership Length Liquid substance Mass Spectrum Analysis Measures Mentorship Metabolism Molecular Molecular Biology Morphology Mus Mutant Strains Mice Mutation Nature Nephrons Nucleotides Onset of illness Open Reading Frames Oxidative Phosphorylation PKD1 gene Pathogenesis Pathway interactions Patients Persons Phenotype Phosphorylation Polycystic Kidney Diseases Positioning Attribute Proteins Proteomics Public Health Publishing Quality of life RNA RNA Binding Reporting Research Research Personnel Resource Sharing Ribonucleoproteins Ribosomal Protein S6 Role Signal Pathway Signal Transduction Techniques Testing Training Translational Research Translations United States Universities Untranslated RNA Wild Type Mouse base career experimental study functional loss in vivo interest kidney cell kidney epithelial cell molecular phenotype mouse genetics mouse model mutant new therapeutic target next generation novel prevent skills therapeutic target therapeutically effective tolvaptan

项目摘要

Polycystic kidney disease (PKD) is characterized by the formation of cysts, which originate from the epithelial tubules of the nephron. Progressive growth of the cysts causes damage and loss of functional nephrons, ultimately leading to end-stage renal failure. The pathophysiology of PKD is incompletely understood, and only one FDA-approved treatment (tolvaptan) exists today. Long noncoding RNAs (lncRNA) – defined by a length >200 nucleotides and absence of a long open reading frame – are a class of non-protein-coding RNAs implicated in a range of diseases. The nature and extent of involvement of lncRNAs in PKD was not previously investigated. Utilizing two independent PKD mouse models, the applicant identified Hoxb3os, a highly conserved lncRNA, which was downregulated in mouse and human ADPKD. Deletion of Hoxb3os in kidney cells resulted in increased phosphorylation of mTOR and its downstream targets. Consistent with activation of mTOR signaling, Hoxb3os mutant cells displayed increased oxidative phosphorylation, increased cell proliferation, and decreased autophagy. The Hoxb3os mutant phenotype was partially rescued upon re- expressing wild-type Hoxb3os in knockout cells. Importantly, deletion of Hoxb3os in wild-type mice recapitulated the in vitro molecular phenotype and resulted in increased mTOR phosphorylation and subsequent increased cell proliferation, and defective autophagy. The overarching hypothesis for this K01 project is that downregulation of Hoxb3os exacerbates cyst formation and/or disease progression by dysregulating multiple pathways, including mTOR signaling. To test this hypothesis, the applicant will decipher the mechanism by which Hoxb3os inhibits mTOR signaling, identify novel Hoxb3os-regulated pathways, and determine the contribution of Hoxb3os to cyst formation in a mouse model of ADPKD. The applicant has assembled an interdisciplinary team of senior investigators to guide the proposed research and provide mentorship during his transition to independence. He will have full access to the University's shared resources (mouse genetics laboratory, high-throughput sequencing, mass spectrometry and proteomics, bioinformatics). The applicant's training plan includes mentorship in core technique and concepts, generating and analyzing data, publishing and presenting results, completing coursework (in mouse genetics, biostatistics, RNA biology, computational methods, bioinformatics), and developing other requisite skills (in leadership, grant proposal development) needed to thrive as an independent investigator. Results from the proposed studies will form the basis for an R01 application to be submitted in the fourth year of this career development award. The applicant's long-term goal is to dedicate his career to advancing basic and translational research on cystic kidney disease as an independent investigator at an academic institution. As a kidney researcher with a deep interest and proven track record in molecular biology, he is uniquely positioned to answer the questions set forth in this proposal and to rapidly advance the mechanistic understanding of lncRNAs in PKD pathogenesis

多囊肾病（PKD）的特点是形成囊肿，囊肿起源于上皮细胞囊肿的进行性生长会导致功能性肾单位的损伤和丧失，最终导致终末期肾衰竭 PKD 的病理生理学尚不完全清楚。目前已有一种 FDA 批准的治疗方法（托伐普坦）——由长度定义。 >200 个核苷酸且缺乏长开放阅读框 – 是一类非蛋白质编码 RNA lncRNA 参与 PKD 的性质和程度此前尚无报道。利用两个独立的 PKD 小鼠模型进行研究，申请人鉴定出 Hoxb3os，这是一种高度的。保守的 lncRNA，在小鼠和人 ADPKD 肾脏中的 Hoxb3os 缺失中下调。细胞导致 mTOR 及其下游靶标的磷酸化增加，这与 mTOR 的激活一致。 mTOR 信号转导，Hoxb3os 突变细胞表现出氧化磷酸化增加，细胞数量增加 Hoxb3os 突变体表型在重新修复后得到部分挽救。在敲除细胞中表达野生型 Hoxb3os 重要的是，在野生型小鼠中删除 Hoxb3os。重现了体外分子表型并导致 mTOR 磷酸化增加随后细胞增殖增加，自噬缺陷。这是 K01 的总体假设。该项目认为，Hoxb3os 的下调会通过以下方式恶化囊肿形成和/或疾病进展：多种途径失调，包括 mTOR 信号传导。为了检验这一假设，申请人将破译。 Hoxb3os 抑制 mTOR 信号传导的机制，识别新的 Hoxb3os 调节途径，以及申请人已确定Hoxb3os对ADPKD小鼠模型中囊肿形成的贡献。组建一个由高级研究人员组成的跨学科团队来指导拟议的研究并提供在向独立过渡期间，他将能够充分利用大学的共享资源。（小鼠遗传学实验室、高通量测序、质谱和蛋白质组学、生物信息学）。申请人的培训计划包括核心技术和概念的指导、生成和分析数据、发布和展示结果、完成课程作业（小鼠遗传学、生物统计学、RNA 生物学、计算方法、生物信息学），以及发展其他必要技能（领导力、拨款提案）发展）作为一名独立研究者的发展所需要的，拟议研究的结果将形成该职业发展奖第四年提交的 R01 申请的基础。申请人的长期目标是致力于推进囊性的基础和转化研究作为一名学术机构的独立研究者，对肾脏疾病有深入的研究。他对分子生物学感兴趣并拥有良好的记录，因此在回答所设问题方面具有独特的优势在此提案中提出并快速推进对 lncRNA 在 PKD 发病机制中的机制理解