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（由申请人提供）：本申请的总体目标是阐明Notch信号传导负调节的不同机制。 Notch 是动物发育中细胞间相互作用和细胞命运规范的主要信号系统中的同名受体。适当调节Notch信号——空间、时间、强度或持续时间——对于正常发育至关重要。此外，Notch 活性异常与许多不同的癌症以及发育、免疫和神经系统疾病有关。因此，拟议的工作对人类健康有很多影响。对发育机制的更深入了解对于开发人类疾病的诊断和治疗工具具有巨大潜力，这是美国国立卫生研究院的核心使命。 我们过去对线虫 LIN-12/Notch 信号传导的研究为 Notch 信号传导的保守作用、机制和调节提供了许多基本见解。我们期望我们在该系统中的持续工作将继续揭示普遍相关的机制，并且选择实验设计的特定方面来最大化这一前景。该应用的每个目标都探索了在教科书发育范例中调节 LIN-12/Notch 活性和稳定性的不同方式，其中六种多能前体细胞通过 LIN-12/Notch 介导的细胞间相互作用采取三种不同命运之一。该图案精确而稳健，反映了对 LIN-12/Notch 活性的严格空间和时间控制以及将 LIN-12/Notch 活性与其他保守信号通路整合的多种模式。 在目标 1 中，我们建议表征我们在负调节因子的靶向筛选中发现的新激酶，以及首先通过癌症患者的突变发现的另一个基因，我们表明它在秀丽隐杆线虫中充当负调节因子。为了评估它们在 Notch 调节中的功能保守性，我们将进行人体细胞测定。在目标 2 中，我们建议研究 LIN-12/Notch 内吞运输及其在发育模式中 EGF 受体活性的负调控。 EGF 受体与 Notch 一样，是一种重要的癌基因，这些通路之间的串扰发生在癌症和正常发育中。在目标 3 中，我们建议阐明活化的核 LIN-12/Notch 在发育模式期间如何被 EGF 受体抑制，以及在响应不利环境的静止期间如何被胰岛素/胰岛素样信号传导抑制。静止是对人类健康至关重要的基本特性，它允许干细胞长时间保持在活性状态，以便在细胞因衰老、损伤或疾病而丧失时能够重新填充组织，而对于癌症来说，静止的干细胞被认为对标准化疗具有抵抗力。根据 NIH 的使命，这些目标将共同提供有关发育和疾病中基本信号通路负调控的许多新信息。