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: 9897729
• 负责人: Emily Margaret Mace
• 金额: $ 6.19万
• 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/9897729
• 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; Stem cells; 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; success

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.

项目摘要 该提议旨在定义人类NK细胞迁移之间知之甚少但关键的关系 和分化。通过整合我们经过验证的人类NK细胞分化系统和高度定量的系统 成像和图像分析，我们将1）定义人类NK细胞分化中迁移的要求， 2）剖析NK细胞发育中间体和基质细胞之间的分子相互作用。 天然杀手（NK）细胞是我们人体防御病毒感染和恶性肿瘤所必需的；他们也是 塑造移植的结果和成功。尽管他们对人类健康的要求有所记录，但 NK细胞的发展和功能的获取知之甚少。与辅助单元的相互作用，例如 淋巴样细胞是人NK细胞发育所必需的，但是这些分子生物学背后的分子生物学 细胞 - 细胞相互作用尚不清楚，也从未被可视化。使用人类NK的体外分化 细胞和定量成像，我们描述了发展中NK细胞和 基质细胞，我们称之为发育突触。此外，我们定义了不同的迁徙 与基质细胞上人NK细胞进行性成熟相关的表型。剖析 NK细胞发育与基质细胞上迁移之间的关系，我们将追求以下特定 目的：1）确定人类NK细胞发育中基质迁移的要求；两者都使用 来自罕见突变的患者的机械约束和NK前体会损害NK细胞的迁移，我们将 定量测量抑制细胞迁移对人NK细胞分化的影响。进一步 描述了粘附和迁移在人类NK细胞发育中发挥作用的作用，我们将：2）定义 基质细胞对NK细胞发育的物理贡献。这将包括评估替代方案 底物确定粘附性和底物刚度对NK细胞发育的贡献。 最后，我们将3）定义NK细胞Uropod和 发展突触。这将描述在细胞中发生的信号转导机理 通过发育突触结合并定义了这与在 迁移细胞。最终目标将在原位和体外进行以使生理发育相关联 与文化形成的突触。 通过这些目标，我们将提高人们对人类NK单元的联系依赖性要求的了解 发展，在过去十年中很少有科学进步的领域。我们的最终目标 是为了确定概括人NK所需的必要和足够的机械和生化信号 细胞发育可有效地产生治疗细胞，而无需使用基质细胞喂食器。通过理解 在此过程中，我们将对细胞迁移的要求产生临床和科学影响的发现。