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

多囊肾脏疾病（PKD）的特征是形成囊肿，囊肿起源于上皮肾单位的管。囊肿的进行性生长会导致功能肾脏的损害和丧失，最终导致末期肾衰竭。 PKD的病理生理学尚不完全理解，只有如今，存在一种由FDA批准的治疗（Tolvaptan）。长的非编码RNA（LNCRNA） - 由长度定义 > 200个核苷酸和长期开放式阅读框架 - 是一类非蛋白质编码RNA 在各种疾病中实施。 LNCRNA参与PKD的性质和程度以前不是调查。利用两种独立的PKD鼠标模型，适用的识别的Hoxb3os，高度保守的lncRNA，在小鼠和人ADPKD中被下调。肾脏中的HOXB3OS删除细胞导致MTOR及其下游靶标的磷酸化增加。与激活一致 MTOR信号传导，HOXB3OS突变细胞显示出增加的氧化磷酸化，细胞增加扩散，并改善自噬。 HOXB3OS突变表型对重新进行了反应在基因敲除细胞中表达野生型HOXB3O。重要的是，在野生型小鼠中删除Hoxb3OS 概括了体外分子表型，导致MTOR磷酸化增加，并且随后增加了细胞增殖和自噬缺陷。该K01的总体假设项目的下调Hoxb3OS的下调加剧了囊肿形成和/或疾病进展功能失调多个途径，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，识别新型HOXB3OS调节途径的机制，并确定Hoxb3OS对ADPKD小鼠模型中囊肿形成的贡献。申请人有组建了一个高级调查员的跨学科团队，以指导拟议的研究并提供在过渡到独立性期间的遗产。他将完全访问大学的共享资源（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精通，生成和分析数据，发布和提出结果，完成课程（在小鼠遗传学，生物统计学，RNA生物学，计算方法，生物信息学）并发展其他必要的技能（在领导力，赠款建议中开发）需要作为独立研究者蓬勃发展。拟议研究的结果将形成在本职业发展奖的第四年提交R01申请的基础。这申请人的长期目标是将自己的职业献给囊性的基础和翻译研究肾脏疾病是学术机构的独立研究者。作为一名肾脏研究员他在分子生物学方面的兴趣和经验证的记录，他在回答问题的问题方面处于独特状态在该提案中的第四，并迅速提高对PKD发病机理中LNCRNA的机械理解