Contact-dependent determinants of human natural killer cell development

人类自然杀伤细胞发育的接触依赖性决定因素

• 批准号: 10315490
• 负责人: Everardo Hegewisch Solloa
• 金额: $ 4.6万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10315490
• 关键词: Actins; Adhesions; Binding; Blocking Antibodies; Bone Marrow; CD3 Antigens; CD34 gene; CRISPR/Cas technology; Cell Count; Cell Differentiation process; Cell Line; Cell Maturation; Cell Proliferation; Cell Survival; Cell physiology; Cells; Coculture Techniques; Cues; Cytoskeleton; Data; Development; Event; Extracellular Matrix; Fetal Liver; Fibronectins; Flow Cytometry; Frequencies; Gene Expression; Genes; Hematopoiesis; Hematopoietic; Human; Immune; Immunotherapy; In Situ; In Vitro; Intestines; Knock-out; Ligands; Light; Liver; Lymphocyte; Lymphoid; Lymphoid Tissue; Mediating; Mesenchymal; Metabolic; Modeling; Mus; Myristica fragrans; NCAM1 gene; Natural Killer Cells; Neural Cell Adhesion Molecules; Neurons; Phenotype; Play; Precursor Natural Killer Cell; Process; Production; Proteins; Proteoglycan; Publishing; Role; Shapes; Signal Pathway; Signal Transduction; Site; Skin; Stromal Cells; Sulfate; Synapses; Testing; Therapeutic; Tissues; Uterus; Viral; Visualization; Work; base; cancer cell; cell motility; cellular imaging; human migration; improved; interest; live cell imaging; migration; nerve stem cell; neural model; receptor; stem cells; tool; transcriptome; transcriptome sequencing; transcriptomics

Abstract: Human natural killer (NK) cells are traditionally identified as CD56+CD3- large granular innate lymphocyte that can detect and eliminate virally infected and malignant cells. Although CD56 is used to identify NK cells, its function is not fully understood. CD56 or neural cell adhesion molecule (NCAM) was first described in neural progenitor cells where it can engage in homotypic and heterophilic interactions with its ligands (e.g., NCAM, fibronectin domains, sulfate proteoglycans) and regulates neuron survival, migration, and development. Interestingly, both xenogenic murine developmentally supportive stroma (e.g., EL08.1D2 and OP9) commonly used for in vitro NK cell differentiation from CD34+ progenitors and stromal cells in sites of hematopoiesis (e.g., bone marrow MSC) express NCAM when compared non-developmentally supportive stroma. Furthermore, work published in the Mace Lab has revealed that inhibition of CD56 (NCAM) mediated interactions between NK cells and stromal cells by a blocking antibody perturbs in vitro NK cell development resulting in decreased NK cell number and migration. I hypothesize that Ncam on developmentally supportive stromal cells and CD56 on NK cells mediate interactions between stromal cells and NK cells that are required for NK cell development, survival and proliferation. My first aim is to characterize the functional role of Ncam expression on developmentally supportive stroma. My preliminary data suggests that the use of Ncam1 knockout murine EL08.1D2 stromal cells for in vitro NK cell differentiation leads to decreased NK cell developmental intermediate survival, proliferation, and migration. I will determine whether Ncam, on developmentally supportive stroma, mediates their ability to support NK cell development, survival, and migration. My second aim is to define the requirement of CD56 on human NK cells for their development and migration. Work by the Mace lab has shown that inhibition of CD56 on NK cells, via blocking antibody, leads to a decrease in cell migration and dysregulation of the actin cytoskeleton. I will delete CD56 in human NK cell progenitors and follow their development in vitro by transcriptomic and phenotypic analysis. Together our study will bring light to a previously unknown CD56 mediated interactions and signaling pathways required for human NK cell proliferation, survival and migration through development.

摘要： 人自然杀伤（NK）细胞传统上被鉴定为CD 56 + CD 3-大颗粒先天性淋巴细胞， 可以检测和清除病毒感染和恶性细胞。虽然CD 56用于识别NK细胞，但其 功能尚未完全理解。CD 56或神经细胞粘附分子（NCAM）首次在神经细胞粘附中被描述。 其中它可以与其配体进行同型和异嗜性相互作用（例如，NCAM， 纤连蛋白结构域、硫酸蛋白聚糖）并调节神经元存活、迁移和发育。 有趣的是，异种鼠发育支持基质（例如，EL08.1D2和OP 9）通常 用于从造血部位的CD 34+祖细胞和基质细胞体外分化NK细胞（例如， 骨髓MSC）表达NCAM。此外，委员会认为， 梅斯实验室发表的工作表明，CD 56（NCAM）的抑制介导了CD 56与CD 56之间的相互作用。 NK细胞和基质细胞通过阻断抗体干扰体外NK细胞发育，导致NK细胞增殖减少。 NK细胞数量和迁移。我假设Ncam对发育支持基质细胞和 NK细胞上的CD 56介导NK细胞活化所需的基质细胞和NK细胞之间的相互作用。 发展、生存和扩散。我的第一个目标是描述NCAM的功能作用 在发育支持性基质上表达。我的初步数据表明，使用Ncam 1 用于体外NK细胞分化的敲除鼠EL08.1D2基质细胞导致NK细胞减少 发育中间存活、增殖和迁移。我将决定是否NCAM，在 发育支持性基质，介导其支持NK细胞发育、存活和 迁移我的第二个目标是确定人类NK细胞发育对CD 56的需求， 迁移梅斯实验室的工作表明，通过阻断抗体抑制NK细胞上的CD 56， 细胞迁移减少和肌动蛋白细胞骨架失调。我将删除人类NK细胞中的CD 56 祖细胞，并通过转录组学和表型分析跟踪它们的体外发育。我们一起 这项研究将揭示以前未知的CD 56介导的相互作用和信号通路 人类NK细胞增殖、存活和迁移所必需的。