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.

项目概要 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影响。在 特别是，已知需要将信使 RNA (mRNA) 亚细胞定位到细胞突起 对于细胞迁移，但在这种情况下 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嵴细胞中表达，这些细胞是 一种特别迁移的细胞类型，产生了脊椎动物头部的大部分以及神经元， 黑色素细胞和侵袭性癌症，包括黑色素瘤。因此，神经嵴细胞提供了丰富的 揭示细胞迁移中 mRNA 定位的分子机制及其相关性的环境 这与哺乳动物的生理学有关。在这个项目中，我将使用培养的黑色素瘤细胞来识别和 表征 mRNA 定位的反式作用调节因子并阐明 mRNA 定位对 蛋白质功能。我还将使用体内小鼠模型来对神经元过程中发生的 mRNA 定位进行分类。 嵴细胞发育并直接测试特征明确的局部 mRNA Kif1c 在过程中的作用 发育和癌症的发生和进展。由于神经嵴和癌症遗传学领域严重依赖 转录研究，此处描述的转录后过程的机械理解可能 对颅面部出生缺陷和黑色素瘤的原因、诊断和治疗提供独特的见解 发病和进展。 我的职业目标是开展一个学术研究项目，分析亚细胞 RNA 的作用和调节 胚胎发生过程中的定位，特别关注神经嵴细胞及其衍生物。我的志向 就是拥有一个能够在单分子水平上确定分子机制并将这些机制联系起来的实验室 体内生理结果的过程。为此，提出的实验和训练计划是 旨在培养尖端技术的专业知识，例如 TIRF 显微镜、计算图像 分析、基因工程和体内疾病测定。这些先进技术的培训将 得到 UT 西南大学资源丰富和协作环境的直接支持，特别是 Gaudenz Danuser 博士、Ondine Cleaver 博士、Khuloud Jaqaman 博士、Lu Le 博士和 Sean 博士的指导 莫里森，我的顾问委员会成员。独立之路奖将提供时间， 资源和自主权来充分开发和启动这一雄心勃勃的研究计划并积累 2024 年秋季启动一个有影响力、蓬勃发展的研究实验室所需的资源、专业知识和经验。