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）：本申请的目标是建立一个有针对性的培训计划，以在获奖期间成功推进我的研究和职业目标。本建议书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，这些技能我几乎没有接触过，例如，有效的granecommunication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的培训。为此，我最近与James Inglese博士的实验室建立了合作关系，他是国家推进转化科学中心（NCATS）检测开发和筛选技术实验室的主任，他将担任我的共同导师，如下所述。这些拟议的培训活动对于我顺利过渡到独立调查员职位是必要的。我的培训的各个方面将由一个咨询委员会的支持，该委员会由五位在校内和校外研究方面具有独特科学和职业专长的顶尖科学家组成。我的研究计划的目标是开发一个创新的研究计划，旨在利用高通量筛选（HTS）库的化学多样性来探索外源配体对RNA结构的调节，这是一种新的方法，我称之为HTS-MoRSEL。通过直接靶向RNA折叠，而不是结合，将鉴定控制几乎任何结构化RNA的折叠/功能的小分子配体。这个高影响力的研究计划对探索RNA生物学不断扩大的领域内的结构-功能关系具有广泛的影响。在获奖期间，我将确定配体调节折叠和功能的几个RNA的生物医学意义。例如，我将检查RNA，调节基因负责病毒复制或致病菌的毒力。此外，我将应用此方法来鉴定能够调节人类致癌microRNA多顺反子结构的配体;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。总之，我的研究结果将为靶向RNA的治疗方法的设计和合成生物学工具的开发提供新的范例。虽然我在博士学位和博士学位上都取得了非凡的研究成功，和博士后研究，我提出的研究计划的新方向需要额外的培训，将建立在我现有的培训。具体来说，最近才被介绍到高温超导化学生物学方法，我正在努力建立我的熟练使用这种方法。鉴于这些检测在我提议的研究计划中的重要性，我需要在奖励期间的指导阶段进行该领域的重点培训。我最近与James Inglese博士建立了合作关系，他同意在这个奖项期间担任共同导师。我将在他的实验室接受重点培训，为我的RNA折叠研究开发适当的HTS测定。在这个职业过渡奖下，我还将在我的导师Ferr-D 'Amar博士（RNA生物物理学和细胞生理学，NHLBI）的实验室接受必要的额外培训，用于在我的HTS研究中确定的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这项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发展培训，以建立一个独立的研究实验室。独特的是，经过这次培训，我将有资格进行HTS测定和RNA-配体相互作用的分子表征。总之，这些综合方法将使我成为一个高素质的候选人，领导一个独立的研究实验室。NHLBI，NCATS和NIH内的环境非常适合该奖项应用程序的各个方面的开发。沿着科学资源，NIH通过由Sharon Milgram博士领导的校内教育和培训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）。此外，NHLBI教育办公室主任赫伯特盖勒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。总之，根据NHLBI职业过渡奖，我将接受研究和职业发展领域的特定培训，我迄今为止几乎没有接触过这些领域，包括但不限于HTS检测开发、指导、教学和实验室管理。在这次培训之后，我将非常有能力实现我的职业目标，即在一所学术大学建立一个高影响力的独立研究实验室。