DEFINING THE ROLE OF CELL MIGRATION IN HUMAN NK CELL DIFFERENTIATION

定义细胞迁移在人类 NK 细胞分化中的作用

• 批准号: 10190802
• 负责人: Emily Margaret Mace
• 金额: $ 37.52万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-02 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10190802
• 关键词: Adhesions; Adhesiveness; Adhesives; Antigen-Presenting Cells; Behavior; Binding; Biochemical; Cell Communication; Cell Differentiation process; Cell Extracts; Cell Line; Cell Maturation; Cell Therapy; Cell surface; Cells; Clinical; Data; Development; Evaluation; Extracellular Matrix; Goals; Health; Human; Image Analysis; Immune; Immune response; Immunotherapy; Impairment; In Situ; In Vitro; Knowledge; Ligands; Lymphocyte Biology; Lymphoid; Lytic; Malignant Neoplasms; Measurement; Measures; Mechanics; Modeling; Molecular; Molecular Biology; Mutation; Natural Killer Cells; Nature; Outcome; Patients; Phenotype; Physical Restraint; Physiological; Play; Precursor Natural Killer Cell; Process; Property; Publishing; Regulation; Resolution; Role; Scientific Advances and Accomplishments; Shapes; Signal Transduction; Site; Stromal Cells; Structure; Supporting Cell; Synapses; System; Technology; Testing; Therapeutic; Transplantation; Virus Diseases; Work; behavior in vitro; cell motility; cellular imaging; design; human imaging; human tissue; in vivo; innovation; mechanotransduction; migration; monolayer; negative affect; novel; quantitative imaging; restraint; stem cells; success; transplantation therapy

PROJECT SUMMARY This proposal aims to define the poorly understood yet critical relationship between human NK cell migration and differentiation. By integrating our validated system of human NK cell differentiation and highly quantitative imaging and image analysis, we will 1) define the requirement for migration in human NK cell differentiation, and 2) dissect the molecular interactions between NK cell developmental intermediates and stromal cells. Natural killer (NK) cells are required for our body's defense against viral infection and malignancy; they also shape the outcome and success of transplantation. Despite their documented requirement in human health, NK cell development and acquisition of function is poorly understood. Interactions with accessory cells such as lymphoid stromal cells are required for human NK cell development, yet the molecular biology behind these cell-cell interactions is not known and has never been visualized. Using in vitro differentiation of human NK cells and quantitative imaging, we have described the contacts formed between developing NK cells and stromal cells, which we termed the developmental synapse. In addition, we have defined distinct migratory phenotypes associated with the progressive maturation of human NK cells on stromal cells. To dissect the relationship between NK cell development and migration on stromal cells, we will pursue the following specific aims: 1) determine the requirement for migration on stroma in human NK cell development; using both mechanical restraint and NK precursors from patients with rare mutations that impair NK cell migration, we will quantitatively measure the effect of inhibiting cell migration on human NK cell differentiation. To further delineate the role that adhesion and migration plays in human NK cell development, we will: 2) define the physical contribution of stromal cells to NK cell development. This will include the evaluation alternative substrates to determine the contribution of adhesiveness and substrate stiffness on NK cell development. Finally, we will 3) define the structural and functional relationship between the NK cell uropod and developmental synapse. This will delineate the mechanism of signal transduction that occurs in cells conjugated through the developmental synapse and define how this is related to the uropod formed in migrating cells. This final aim will be performed in situ and in vitro to correlate the physiological developmental synapse with that formed in culture. Through these aims we will advance our knowledge of the contact-dependent requirements for human NK cell development, a field in which few scientific advances have been made in the past ten years. Our ultimate goal is to identify necessary and sufficient mechanical and biochemical signals required to recapitulate human NK cell development to efficiently generate therapy cells without the use of stromal cell feeders. By understanding the requirement for cell migration in this process, we will generate findings of both clinical and scientific impact.

项目总结 这项提议旨在定义人类NK细胞迁移之间知之甚少但却至关重要的关系 和差异化。通过集成我们验证的人类NK细胞分化系统和高度定量的 成像和图像分析，我们将1)定义人类NK细胞分化中的迁移要求， 2)分析NK细胞发育中间体与基质细胞之间的分子相互作用。 自然杀伤(NK)细胞是我们身体抵御病毒感染和恶性肿瘤所必需的；它们还 塑造移植的结果和成功。尽管它们对人类健康的要求有据可查， NK细胞的发育和获得功能知之甚少。与附属细胞的相互作用，如 淋巴基质细胞是人类NK细胞发育所必需的，但这些细胞背后的分子生物学 细胞之间的相互作用是未知的，也从未被可视化。人NK细胞体外分化的应用 细胞和定量成像，我们已经描述了发育中的NK细胞和 基质细胞，我们称之为发育性突触。此外，我们还定义了不同的迁徙 与人NK细胞在基质细胞上逐渐成熟相关的表型。要仔细分析 NK细胞发育与基质细胞迁移的关系，我们将追求以下特异性 目的：1)确定人类NK细胞发育中基质迁移的要求；使用两者 机械约束和NK前体来自罕见突变损害NK细胞迁移的患者，我们将 定量检测抑制细胞迁移对人NK细胞分化的影响。为了进一步 描述黏附和迁移在人类NK细胞发育中的作用，我们将：2)定义 基质细胞对自然杀伤细胞发育的生理贡献。这将包括评价备选方案。 以确定粘附性和底物硬度对NK细胞发育的贡献。 最后，我们将3)确定NK细胞尾足和尾足之间的结构和功能关系 发育性突触。这将描述发生在细胞中的信号转导机制 通过发育中的突触共轭，并定义这与在 迁移细胞。这一最终目标将在原位和体外进行，以关联生理发育 与在文化中形成的突触。 通过这些目标，我们将促进我们对人类NK细胞接触依赖要求的了解 发展，这是一个在过去十年中几乎没有取得科学进展的领域。我们的终极目标 是确定必要和足够的机械和生化信号，以重述人类NK细胞 细胞培养，无需使用基质细胞饲养器，即可高效产生治疗细胞。通过理解 在这个过程中，细胞迁移的要求，我们将产生临床和科学影响的结果。