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: 10676832
• 负责人: Everardo Hegewisch Solloa
• 金额: $ 2.73万
• 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 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10676832
• 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; Phenotype; Play; Precursor Natural Killer Cell; Process; Production; Proliferating; Proteins; Proteoglycan; Publishing; Role; Shapes; Signal Pathway; Signal Transduction; Site; Skin; Stromal Cells; Sulfate; Synapses; Testing; Therapeutic; Tissues; Uterus; Viral; Visualization; Work; cancer cell; cell fixing; cell motility; cellular imaging; human migration; improved; interest; live cell imaging; migration; nerve stem cell; neural model; neuronal survival; 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.

抽象的： 传统上，人类天然杀手（NK）细胞被识别为CD56+CD3-大的颗粒状淋巴细胞 可以检测并消除病毒感染和恶性细胞。尽管CD56用于识别NK细胞，但 功能尚未完全理解。首先在神经中描述了CD56或神经细胞粘附分子（NCAM） 祖细胞可以与其配体进行同型和异质相互作用（例如，NCAM， 纤连蛋白结构域，硫酸盐蛋白聚糖）并调节神经元的存活，迁移和发育。 有趣的是，特种鼠在发育中的辅助基质（例如EL08.1D2和OP9）都通常 用于体外NK细胞与CD34+祖细胞和基质细胞分化的造血部位（例如， 当比较非发育支持基质时，骨髓MSC）表达NCAM。此外， 在MACE实验室发表的工作表明，抑制CD56（NCAM）介导的相互作用 NK细胞和基质细胞通过阻断抗体在体外NK细胞发育中导致下降 NK细胞编号和迁移。我假设NCAM在发育中支持性的基质细胞和 NK细胞上的CD56介导NK细胞所需的基质细胞和NK细胞之间的相互作用 发展，生存和增殖。我的第一个目的是表征NCAM的功能作用 关于发育支持性基质的表达。我的初步数据表明使用NCAM1 用于体外NK细胞分化的敲除鼠EL08.1D2基质细胞导致NK细胞降低 发展中间生存，增殖和迁移。我将确定ncam是否在 发育支持基质，介导其支持NK细胞开发，生存和 迁移。我的第二个目的是确定CD56对人NK细胞开发的需求， 迁移。 MACE实验室的工作表明，通过阻断抗体，CD56对NK细胞的抑制导致 肌动蛋白细胞骨架的细胞迁移和失调减少。我将在人类NK单元中删除CD56 祖细胞并通过转录组和表型分析在体外遵循其发育。在一起 研究将使先前未知的CD56介导的相互作用和信号通路的光线 人类NK细胞增殖，生存和通过发育迁移所必需的。