Regulation and function of subcellular RNA localization in neural crest cells and their derivatives

神经嵴细胞及其衍生物亚细胞RNA定位的调控和功能

• 批准号: 10739280
• 负责人: Megan L Norris
• 金额: $ 13.24万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-20 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739280
• 关键词: Address; Advisory Committees; Affect; Attention; Award; Biological Assay; Catalogs; Cell physiology; Cells; Co-Immunoprecipitations; Congenital Abnormality; Data; Defect; Development; Diagnosis; Disease; Elements; Embryonic Development; Environment; Galium; Genes; Genetic Engineering; Genetic Transcription; Goals; Head; Human; Image Analysis; In Vitro; Individual; Intervention; Invaded; Kinesin; Learning; Link; Location; Malignant Neoplasms; Mammalian Cell; Melanoma Cell; Mentorship; Messenger RNA; Modeling; Molecular; Motor; Mus; Mutation; Neural Crest; Neural Crest Cell; Neurologic; Neurons; Outcome; Pathway interactions; Phenotype; Physiological; Physiology; Post-Transcriptional Regulation; Prevention; Process; Proteins; RNA; Regulation; Research; Resources; Role; Route; Running; Schwann Cells; System; Techniques; Technology; Testing; Time; Tissues; Total Internal Reflection Fluorescent; Training; Trans-Activators; Transcription Process; Translations; cancer genetics; career; cell behavior; cell motility; cell type; collaborative environment; craniofacial; design; experience; experimental study; falls; gain of function mutation; in vivo; insight; interest; light microscopy; loss of function; melanocyte; melanoma; migration; mouse model; novel; posttranscriptional; programs; protein function; protein protein interaction; single molecule; trafficking; transcriptomics; tumor progression

PROJECT SUMMARY The long-term objective of this project is to determine the general principles and causal mechanisms underlying subcellular RNA localization, and the physiological implications during development and disease. In particular, the subcellular localization of messenger RNA (mRNA) to cell protrusions is known to be required for cell migration, but the mechanisms by which mRNA localization regulates protein function in this setting are unclear. Many genes implicated in mRNA localization are expressed in neural crest cells, which are a particularly migratory cell type that gives rise to much of the vertebrate head, as well as neurons, melanocytes, and aggressive cancers including melanoma. As such, neural crest cells provide a rich environment to uncover molecular mechanisms of mRNA localization in cell migration as well as the relevance there of to mammalian physiology. In this project, I will use cultured melanoma cells to identify and characterize trans-acting regulators of mRNA localization and elucidate the role of mRNA localization on protein function. I will also use in vivo mouse models to catalog mRNA localization that occurs during neural crest cell development and directly test the role of a well characterized localized mRNA, Kif1c, during development and cancer onset and progression. As neural crest and cancer genetics fields rely heavily on transcriptional studies, the mechanistic understanding of a post-transcriptional process as described here may provide unique insight into the cause, diagnosis and treatment of craniofacial birth defects and melanoma onset and progression. My career goal is to run an academic research program analyzing the role and regulation of subcellular RNA localization during embryogenesis, with special attention to neural crest cells and their derivatives. My ambition is to have a lab that can determine molecular mechanisms at the single-molecule level and causally link those processes to physiological outcomes in vivo. To this end, the proposed experiments and training plan are designed to develop expertise in cutting-edge technologies such as TIRF microscopy, computational image analysis, genetic engineering and in vivo disease assays. Training in these advanced techniques will be directly supported by the resource-rich and collaborative environment at UT Southwestern, and especially mentorship from Dr. Gaudenz Danuser, Dr. Ondine Cleaver, Dr. Khuloud Jaqaman, Dr. Lu Le and Dr. Sean Morrison, who make up my advisory committee. The Pathway to Independence Award will provide the time, resources and autonomy to fully develop and initiate this ambitious research program and accrue the resources, expertise and experience necessary to launch an impactful, thriving research lab in Fall 2024.

项目摘要 该项目的长期目标是确定一般原则和因果机制 潜在的亚细胞RNA定位以及发育和疾病期间的生理意义。在 特别是，已知允许信使RNA（mRNA）的亚细胞定位已知需要 对于细胞迁移，但是在这种情况下mRNA定位调节蛋白质功能的机制是 不清楚。许多与mRNA定位有关的基因在神经rest细胞中表达， 一种特别迁移的细胞类型，引起大部分脊椎动物头以及神经元， 黑色素细胞和包括黑色素瘤在内的侵略性癌症。因此，神经rest细胞提供丰富的 环境发现细胞迁移中mRNA定位的分子机制以及相关性 哺乳动物生理学。在这个项目中，我将使用培养的黑色素瘤细胞识别和 表征mRNA定位的跨作用调节剂并阐明mRNA定位在 蛋白质功能。我还将使用体内小鼠模型来分类神经期间发生的MRNA定位 波峰细胞的发育并直接测试良好表征的局部mRNA KIF1C的作用 发展和癌症发作和进展。随着神经rest和癌症遗传学领域的严重依赖 转录研究，对此处所述的转录后过程的机理理解可能 提供有关颅面先天缺陷和黑色素瘤的原因，诊断和治疗的独特见解 发作和进展。 我的职业目标是开展一项学术研究计划，分析亚细胞RNA的角色和调节 胚胎发生过程中的定位，并特别注意神经rest细胞及其衍生物。我的野心 是要有一个可以在单分子水平上确定分子机制并因果关系的实验室 体内生理结果的过程。为此，拟议的实验和培训计划是 旨在发展诸如TIRF显微镜，计算图像之类的尖端技术方面的专业知识 分析，基因工程和体内疾病分析。这些高级技术的培训将是 在UT西南部的资源丰富和协作环境中直接支持，尤其是 Gaudenz Danuser博士，Ondine Cleaver博士，Khuloud Jaqaman博士，Lu Le博士和Sean博士的指导 组成我的咨询委员会的莫里森。获得独立奖的途径将提供时间， 充分制定和启动这项雄心勃勃的研究计划的资源和自主权，并累积 资源，专业知识和经验在2024年秋季推出一个有影响力的蓬勃发展的研究实验室。