Regulating RNA function by modulating RNA folding with exogenous ligands

通过外源配体调节 RNA 折叠来调节 RNA 功能

• 批准号: 9119049
• 负责人: Nathan Baird
• 金额: $ 24.37万
• 依托单位: UNIVERSITY OF THE SCIENCES PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9119049
• 关键词: Advisory Committees; Affect; Area; Award; Bacterial Genes; Binding; Biochemical; Biological Assay; Biological Models; Biological Process; Biology; Biophysics; Calorimetry; Career Mobility; Career Transition Award; Catalytic RNA; Cell physiology; Cells; Charge; Chemicals; Collaborations; Complex; Development; Divalent Cations; Doctor of Philosophy; Drug Targeting; Education; Educational process of instructing; Effectiveness; Environment; Exposure to; Extramural Activities; Fluorescence; Fluorescence Resonance Energy Transfer; Genes; Genetic Transcription; Glutamine; Glycine; Goals; HIV-1; Head; Human; Hydrogen Bonding; Laboratories; Laboratory Research; Lead; Libraries; Ligand Binding; Ligands; Measures; Mediating; Mentors; Methodology; Methods; MicroRNAs; Modeling; Molecular; Monitor; Morphologic artifacts; Mutation; National Heart, Lung, and Blood Institute; Oncogenic; Pathway interactions; Phase; Positioning Attribute; Postdoctoral Fellow; Process; Professional Competence; Property; Qualifying; RNA; RNA Binding; RNA Folding; RNA Interference; Reading; Regulator Genes; Reporter; Research; Research Personnel; Resources; Roentgen Rays; Role; Scientist; Shapes; Site; Structure; Structure-Activity Relationship; Testing; Titrations; Training; Training Activity; Training and Education; Transcript; Translational Research; United States National Institutes of Health; Universities; Ursidae Family; Viral Genome; Virulence; Virus Replication; Work; X-Ray Crystallography; assay development; base; career; career development; design; drug development; fluorophore; high throughput screening; in vivo; innovation; instrumentation; laboratory equipment; novel; novel therapeutics; nucleocytoplasmic transport; pathogenic bacteria; programs; research and development; research study; screening; small molecule; small molecule libraries; success; synthetic biology; therapeutic target; tool; transcription termination

DESCRIPTION (provided by applicant): The goal of this application is to establish a focused training plan to successfully advance my research and career goals during the award period. The details provided in this proposal include specific action steps for me to gain training in career skills that I have had little exposure to that include, for example, effective grantsmanship mentoring, and laboratory management. Furthermore, to realize my research goals, I am in need of training in new scientific fields, those of high-throughput screening assays and chemical biology. To that end, I have recently established a collaboration with the laboratory of Dr. James Inglese, director of the Assay Development and Screening Technologies Laboratory in the National Center for Advancing Translational Sciences (NCATS), who will serve as my co-mentor as described below. These proposed training activities are necessary to prepare me for a successful transition to an independent investigator position. All aspects of my training will be supported by an Advisory Committee comprised of five leading scientists with distinct scientific and career expertise in both intramural and extramural research. The goal of my research program is to develop an innovative research plan aimed at leveraging the chemical diversity of high-throughput screening (HTS) libraries to explore the modulation of RNA structure by exogenous ligands, a novel methodology I refer to as HTS-MoRSEL. By targeting RNA folding directly, rather than binding, small molecule ligands will be identified that control the folding/function of nearly any structured RNA. This high-impact research program has broad implications for probing structure-function relationships within the expanding field of RNA biology. During the award period, I will identify ligands modulating the folding and function of several RNAs of biomedical significance. For example, I will examine RNAs that regulate genes responsible for viral replication or virulence in pathogenic bacteria. Additionally, I will apply tis methodology to identify ligands capable of modulating the structure of a human oncogenic microRNA polycistron; the structure and folding of this large RNA transcript are responsible for autoregulating its processing by the RNAi pathway machinery. Together, my results will present a new paradigm for the design of therapeutics targeting RNA and the development of synthetic biology tools. While I have demonstrated exceptional research success both in my Ph.D. and post-doctoral studies, the new direction of my proposed research plan necessitates additional training that will build on my existing training. Specifically, only having been introduced recentl to HTS chemical biology methodologies, I am working to establish my proficiency employing this approach. Given the prominence of these assays in my proposed research program, I am in need of focused training in this field during the mentored phase of the award period. I recently established a collaboration with Dr. James Inglese, who has agreed to serve as co-mentor during this award. I will receive focused training in his laboratory towards the development of appropriate HTS assays for my RNA folding studies. Under this Career Transition Award I will also receive necessary additional training in the lab of my mentor, Dr. Ferr�-D'Amar� (RNA Biophysics and Cellular Physiology, NHLBI) for the molecular and atomistic characterization studies of RNA-ligand interactions identified in my HTS studies. The existing state-of-the-art instrumentation resources within these labs are more than sufficient to complete all aspects of this proposed research program. Both Dr. Ferr�-D'Amar� and Dr. Inglese will work together to provide me with necessary career development training for establishing an independent research lab. Uniquely, following this training I will be aptly qualified to perform both HTS assay and molecular characterization of RNA-ligand interactions. Together, these combined approaches will make me a highly qualified candidate to lead an independent research laboratory. The environment within the NHLBI, NCATS, and NIH are exceptionally suited for the development of all aspects of this award application. Along with the scientific resources, the NIH offers extensive career development programs for post-doctoral researchers (e.g. the Career Advancement Toolkit Track) through the Office of Intramural Education and Training, headed by Dr. Sharon Milgram. Furthermore, Dr. Herbert Geller, director of the NHLBI Office of Education, offers many complementary resources and will provide me with individualized career development guidance. In summary, under the NHLBI Career Transition Award, I will receive specific training in research and career development areas in which I have had little exposure to date including, but not limited to, HTS assay development, mentoring, teaching, and lab management. Subsequent to this training I will be excellently positioned to achieve my career goal of establishing a high-impact, independent research laboratory at an academic university.

描述（由申请人提供）：本申请的目标是制定一个有针对性的培训计划，以在奖励期间成功推进我的研究和职业目标。本提案中提供的详细信息包括我获得职业技能培训的具体行动步骤。我很少接触过这些领域，例如有效的资助指导和实验室管理，此外，为了实现我的研究目标，我需要在新的科学领域、高通量筛选分析和化学生物学领域接受培训。为此，我有。最近与国家转化科学促进中心 (NCATS) 检测开发和筛选技术实验室主任 James Inglese 博士的实验室建立了合作，他将担任我的共同导师，如下所述。我的研究目标是由五位在校内和校外研究领域具有独特科学和职业专业知识的领先科学家组成的咨询委员会为我成功过渡到独立研究者职位做好准备。计划是制定一项创新研究计划，旨在利用高通量筛选（HTS）文库的化学多样性来探索外源配体对 RNA 结构的调节，我将这种新方法称为 HTS-MoRSEL，通过直接靶向 RNA 折叠，而不是。将鉴定出控制几乎所有结构化 RNA 的折叠/功能的结合小分子配体。识别配体调节几种具有生物医学意义的 RNA 的折叠和功能，例如，我将研究调节负责病原细菌中病毒复制或毒力的基因的 RNA。此外，我将应用这种方法来识别能够调节人类结构的配体。致癌的 microRNA 多顺反子；这种大 RNA 转录物的结构和折叠负责通过 RNAi 途径机制自动调节其加工，我的研究结果将为靶向 RNA 的治疗设计和合成的开发提供一个新的范例。虽然我在博士和博士后研究中都取得了非凡的研究成功，但我提出的研究计划的新方向需要在我现有的培训基础上进行额外的培训。 HTS 化学生物学方法，我正在努力建立我使用这种方法的熟练程度，鉴于这些分析在我提出的研究计划中的突出地位，我需要在我最近设立的奖励期间进行该领域的集中培训。与合作James Inglese 博士同意在该奖项期间担任联合导师，我将在他的实验室接受集中培训，为我的 RNA 折叠研究开发适当的 HTS 检测方法。在我的导师 Ferr�-D'Amar� 博士（RNA 生物物理学和细胞生理学，NHLBI）的实验室接受培训，进行我的 HTS 研究中发现的 RNA-配体相互作用的分子和原子表征研究。这些实验室内现有的最先进的仪器资源足以完成该拟议研究计划的各个方面，Ferr�-D'Amar� 博士和 Inglese 博士将共同努力为我提供必要的职业生涯。独特的是，经过本次培训，我将具备执行 HTS 测定和 RNA-配体相互作用的分子表征的资格，这些组合方法将使我成为领导独立研究的合格候选人。实验室内的环境。 NHLBI、NCATS 和 NIH 非常适合该奖项申请的各个方面的发展，除了科学资源外，NIH 还通过办公室为博士后研究人员提供广泛的职业发展计划（例如职业发展工具包轨道）。由 Sharon Milgram 博士领导的校内教育和培训学院此外，NHLBI 教育办公室主任 Herbert Geller 博士提供了许多补充资源，并将为我提供个性化的职业发展指导。 NHLBI 职业转型奖，我将接受迄今为止我很少接触过的研究和职业发展领域的具体培训，包括但不限于 HTS 检测开发、指导、教学和实验室管理。能够很好地实现我的职业目标，即在一所学术大学建立一个高影响力的独立研究实验室。