New modes and mechanisms of negative regulation of LIN-12/Notch in C. elegans

线虫中LIN-12/Notch负调控的新模式和新机制

• 批准号: 9028823
• 负责人: Iva S Greenwald
• 金额: $ 34.3万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-21 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9028823
• 关键词: Address; Adopted; Aging; Animals; Binding; Biochemical; Biological Assay; Caenorhabditis elegans; Cancer Etiology; Cancer Patient; Candidate Disease Gene; Cell Communication; Cell Culture Techniques; Cells; Complex; Development; Diagnostic; Disease; Down-Regulation; Endocytosis; Ensure; Environment; Epidermal Growth Factor Receptor; Experimental Designs; Feedback; Gene Targeting; Genes; Genetic; Goals; Health; Homeostasis; Human; Human Development; Image; Immune System Diseases; Individual; Injury; Insulin; Knowledge; Larva; Lead; Life; Ligands; Lobular Neoplasia; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane; Mission; Mutate; Mutation; Nuclear; Oncogenes; Oncogenic; Pathway interactions; Pattern; Phosphotransferases; Physiology; Positioning Attribute; Property; RNA Interference; Receptor Activation; Refractory; Regulation; Reporter Genes; Resistance; Role; Signal Pathway; Signal Transduction; Somatomedins; Spottings; Stem cells; Surface; Syndrome; System; Tertiary Protein Structure; Textbooks; Therapeutic; Time; Tissues; Trans-Activators; Transgenes; Tumor Suppressor Proteins; United States National Institutes of Health; Work; base; cell fate specification; chemotherapy; developmental disease; gene discovery; genetic analysis; human disease; in vivo; in vivo imaging; insight; mutant; nervous system disorder; notch protein; novel strategies; precursor cell; prevent; public health relevance; receptor; response; stem; tool; trafficking; transcription factor; transcriptomics

 DESCRIPTION (provided by applicant): The overall goal of this application is to elucidate different mechanisms for negative regulation of Notch signaling. Notch is the eponymous receptor in a major signaling system for cell-cell interactions and cell fate specification in animl development. Regulating Notch signaling appropriately--in space, time, strength or duration--is critically important for normal development. Furthermore, aberrant Notch activity has been implicated in many different cancers, as well as in developmental, immune, and neurological disorders. Thus, the proposed work has many implications for human health. The deeper understanding of developmental mechanism has great potential for developing diagnostic and therapeutic tools for human disease, a central mission of the NIH. Our past work on LIN-12/Notch signaling in C. elegans has afforded many fundamental insights into conserved roles, mechanisms, and regulation of Notch signaling. We expect that our continued work in this system will continue to reveal mechanisms of general relevance, and particular aspects of experimental design were chosen to maximize this prospect. Each aim of this application explores different ways LIN-12/Notch activity and stability is modulated in a textbook developmental paradigm in which six multipotential precursor cells adopt one of three distinct fates through LIN-12/Notch-mediated cell-cell interactions. The patterning is precise and robust, reflecting tight spatial and temporal control of LIN-12/Notch activity and multiple modes for integrating LIN-12/Notch activity with other conserved signaling pathways. In Aim 1, we propose to characterize new kinases we identified in a targeted screen for negative regulators and another gene, first identified by a mutation in a cancer patient, we showed acts as a negative regulator in C. elegans. To assess their functional conservation in Notch regulation, we will perform a human cell assay. In Aim 2, we propose to investigate LIN-12/Notch endocytic trafficking and its negative regulation by EGF Receptor activity in developmental patterning. EGF Receptor, like Notch, is an important oncogene, and crosstalk between these pathways occurs in cancer as well as in normal development. In Aim 3, we propose to elucidate how activated nuclear LIN-12/Notch is inhibited by EGF receptor during developmental patterning and by Insulin/Insulin-like Signaling during quiescence in response to unfavorable environment. Quiescence is a fundamental property of critical importance for human health, allowing stem cells to persist over prolonged periods in a competent state so as to be available to repopulate tissues when cells are lost to aging, injury or disease, and for cancer, quiescent stem-like cells are believed to be resistant to standard chemotherapy. Together, these aims will provide much new information about negative regulation of a fundamental signaling pathway in development and disease, in accord with the mission of the NIH.

 描述（由应用程序提供）：此应用程序的总体目标是阐明对Notch信号负面调节的不同机制。 Notch是主要信号系统中用于细胞 - 细胞相互作用和动画开发中细胞脂肪规范的有色接收器。适当地调节Notch信号 - 空间，时间，强度或持续时间 - 至关重要。此外，在许多不同的癌症以及发育，免疫学和神经系统疾病中已经实施了异常的缺口活动。这是对人类健康的许多影响。对发展机制的更深入的理解具有开发人类疾病的诊断和治疗工具的巨大潜力，这是NIH的核心任务。我们过去在秀丽隐杆线虫中对LIN-12/Notch信号传导的工作为保守的作用，机制和Notch信号调节提供了许多基本见解。我们希望我们在该系统中的继续工作将继续揭示一般相关性的机制，并选择了实验设计的特定方面以最大程度地提高该前景。该应用程序的每个目标探讨了LIN-12/Notch活动和稳定性的不同方式，在教科书的发展范式中调节，在该范式中，通过LIN-12/Notch介导的细胞细胞相互作用，六个多能前体细胞采用三个不同的命运之一。模式是精确且健壮的，反映了LIN-12/Notch活性的紧密空间和临时控制，以及多种将LIN-12/Notch活性与其他组成的信号通路集成的多种模式。在AIM 1中，我们建议表征我们在靶向调节剂和另一个基因的靶向筛选中鉴定出的新激酶，最初是由癌症患者突变鉴定出来的，我们表现为秀丽隐杆线虫中的负调节剂。为了评估其在Notch调节中的功能保护，我们将执行人类细胞测定法。在AIM 2中，我们建议研究LIN-12/Notch内吞交易及其对EGF受体活性在发育模式中的负调控。像Notch一样，EGF受体是重要的癌基因，这些途径之间的串扰在癌症以及正常发育中发生。在AIM 3中，我们建议阐明在发育模式期间EGF受体抑制活化的核LIN-12/NOTCH以及在静止期间胰岛素/胰岛素样信号在响应不利的环境中如何抑制。静止是对人类健康至关重要的基本特性，可以使干细胞在合理状态下长时间持续存在，以便在细胞失去衰老，损伤或疾病时可以重新填充组织，并且对癌症，静止干细胞被认为是对标准化学疗法的抗性。总之，这些目标将根据NIH的任务提供有关开发和疾病基本信号通路的负面调节的许多新信息。