Defining the role of Drip27, a novel long noncoding RNA, in erythropoiesis

定义 Drip27（一种新型长非编码 RNA）在红细胞生成中的作用

基本信息

批准号：
8968039
负责人：
Vikram R. Paralkar
金额：
$ 15.68万
依托单位：
UNIVERSITY OF PENNSYLVANIA
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：
2015-07-27 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8968039
关键词：
Advisory Committees Anemia Area Binding Bioinformatics Biological Biological Models Biological Process Biology Biotin Cell Count Cell Cycle Cell Cycle Arrest Cell Cycle Kinetics Cell Cycle Progression Cell Cycle Stage Cell Line Cell division Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Complex Cyclin-Dependent Kinases DNA Data Development Development Plans Disease Dysmyelopoietic Syndromes ES Cell Line Environment Erythroblasts Erythrocytes Erythroid Erythroid Cells Erythropoiesis Faculty Flow Cytometry Foundations Future Gene Expression Gene Targeting Genes Genetic Genetic Materials Genetic Transcription Genomics Goals Grant Hematological Disease Hematology Hematopoiesis Hematopoietic Hematopoietic Neoplasms Human Image Introns Knockout Mice Knowledge Laboratories Laboratory Research Length Longitudinal Studies Malignant - descriptor Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Mediator of activation protein Mentors Mentorship Messenger RNA Modeling Molecular Molecular Conformation Mus Myeloproliferative disease Names Pattern Pediatric Hospitals Pennsylvania Philadelphia Physicians Physiological Play Polyadenylation Positioning Attribute Process Protein Binding Proteins Publishing RNA RNA Sequences RNA Splicing RNA-Binding Proteins RNA-Protein Interaction Reporting Research Role Saint Jude Children&apos s Research Hospital Scientist Signal Transduction Site Spleen Staining method Stains Structure Techniques Technology Testing Training Transcript Transcription Initiation Site Universities Untranslated RNA Western Blotting Work base career development embryonic stem cell experience inhibitor/antagonist insight novel oncology public health relevance single molecule skills symposium tool transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): This application describes the candidate's research on the genetics of red blood cell development (erythropoiesis) to be performed within the context of a 5-year mentored career development plan. His ultimate goal is to become an independent physician-scientist in the area of laboratory-based, academic hematology- oncology. Under the guidance of his primary research mentor Dr. Gerd Blobel at the Children's Hospital of Philadelphia (CHOP), and his co-mentor Dr. Mitchell Weiss at St. Jude Children's Research Hospital, Dr. Paralkar has developed a structured training plan consisting of intensive laboratory research, coursework in RNA biology and bioinformatics, regular participation at scientific conferences, and oversight by an experienced faculty advisory committee. Drs. Blobel and Weiss have extensive experience in training scientists and physician-scientists, and the educational/training environments at CHOP and the University of Pennsylvania are outstanding. Dr. Paralkar's laboratory work centers on long non-coding RNAs (LncRNAs), a recently appreciated class of genetic material that modulates gene expression and regulates diverse aspects of normal and pathological mammalian development. Thousands of LncRNAs are believed to exist, but only few have been studied in detail. Dr. Paralkar has identified a novel LncRNA named Drip27 that regulates the critical cell cycle inhibitor p27. This proposal investigates the hypothesis that Drip27 LncRNA directly activates p27 transcription, and thereby regulates cell division and replication during erythropoiesis. The long term goal of this research is to define the role of Drip27 in normal hematopoietic development and associated diseases such as blood cancers. In preliminary studies, Dr. Paralkar used comprehensive RNA-Sequencing and bioinformatic tools to identify hundreds of novel LncRNAs in mouse and human erythroid precursors. He chose a novel conserved LncRNA named `Drip27,' and he used CRISPR technology to delete it in an erythroid cell line. Deletion led to a significant reduction i the expression of the crucial cell cycle inhibitor p27. This proposal aims to study the effect of Drip27 transcript loss on erythropoiesis by generating mouse ES cells with truncated Drip27 transcript and differentiating them into erythroblasts (Aim 1). The mechanisms by which Drip27 functions will be examined by imaging the subcellular localization of Drip27 RNA molecules and by assessing for the presence of chromosomal looping between the Drip27 and the p27 loci (Aim 2). Published work demonstrates that many LncRNAs regulate gene expression through physical interactions with transcription factors/co-factors/chromatin modifying complexes that orchestrate global gene expression. To further define the mechanism by which Drip27 acts on the p27 locus, proteins binding to Drip27 will be identified (Aim 3). These three aims, put together, will establish the role played by Drip27 in erythropoiesis and the mechanism by which it regulates its target gene. If successful, this work will illuminate novel regulatory mechanisms in erythropoiesis and paradigms for noncoding RNA function. In turn, these findings are likely to produce insights into various blood diseases including anemias, myeloproliferative disorders and myelodysplastic syndromes. Performing this research in the context of a rigorous formal training plan within a highly supportive academic enviroment will provide the candidate essential data and professional tools for advancement into an independent position as a research oriented academic physician-scientist.

描述（由应用程序提供）：本应用程序描述了候选人关于在为期5年的指导职业发展计划的背景下进行的关于红细胞发育（erythropoiesis）遗传学（erythropoiesis）的研究。他的最终目标是成为基于实验室的学术血液肿瘤学领域的独立身体科学家。在他在费城儿童医院（CHOP）的主要研究导师Gerd Blobel博士的指导下，以及他在圣裘德儿童研究医院的Mitchell Weiss博士的指导，Paralkar博士制定了一项结构化培训计划，该计划由RNA生物学和BioInformics的课程研究，经验丰富的委员会，经验丰富的委员会，并经验丰富的委员会，并与众不同地委员会。 Blobel博士和Weiss博士在培训科学家和医师科学家方面拥有丰富的经验，Chop和Pennsylvania大学的教育/培训环境非常出色。 Paralkar博士的实验室工作集中于长期非编码RNA（LNCRNA），这是一种最近欣赏的遗传物质，可调节基因表达并调节正常和病态哺乳动物发育的潜水员方面。据信数千个LNCRNA存在，但只有很少的研究详细研究了。 Paralkar博士的实验室工作集中于长期非编码RNA（LNCRNA），这是一种最近欣赏的遗传物质，可调节基因表达并调节正常和病态哺乳动物发育的潜水员方面。据信数千个LNCRNA存在，但只有很少的研究详细研究了。 Paralkar博士确定了一种名为DRIP27的新型lncrna，该LNCRNA调节关键细胞周期抑制剂p27。该提案调查了以下假设：滴滴27 lncRNA直接激活p27转录，从而调节红细胞生成期间的细胞分裂和复制。这项研究的长期目标是确定滴滴27在正常造血性发育和相关疾病（如血液癌）中的作用。在初步研究中，Paralkar博士使用了全面的RNA测序和生物信息学工具来识别小鼠和人类红细胞前体中数百种新型LNCRNA。他选择了一种名为“ Drip27”的小说构成的lncrna，他使用CRISPR技术在红系细胞系中删除了它。缺失导致显着降低I的表达至关重要的细胞周期抑制剂p27。该提案旨在通过生成带有截短的滴滴27转录物的小鼠ES细胞并将其区分成粉红细胞的小鼠ES细胞来研究DRIP27转录物损失对红细胞生成的影响（AIM 1）。通过对DRIP27 RNA分子的亚细胞定位以及评估滴水27和P27局部染色体环的存在（AIM 2），将检查滴水27函数的机制（AIM 2）。已发表的工作表明，许多LNCRNA通过与转录因子/co因子/染色质修饰的复合物的物理相互作用来调节基因表达，从而调节了策划全球基因表达的复合物。为了进一步定义DRIP27在P27基因座上作用的机制，将鉴定出蛋白质与DRIP27的结合（AIM 3）。总结的这三个目标将确定Drip27在红细胞生成中的作用以及调节其靶基因的机制。如果成功的话，这项工作将阐明红细胞生成的新型调节机制和非编码RNA功能的范例。反过来，这些发现可能会对各种血液疾病产生见解，包括贫血，骨髓增生性疾病和骨髓增生综合症。在高度支持的学术环境中进行严格的正式培训计划的背景下，将提供这项研究，将为候选人的基本数据和专业工具提供晋升为独立职位，作为研究为导向研究的学术医师科学家。