Contact-dependent determinants of human natural killer cell development

摘要： 人类自然杀伤(NK)细胞传统上被认为是CD56+CD3-大颗粒天然淋巴细胞， 可检测并清除病毒感染和恶性细胞。虽然CD56被用来识别NK细胞，但其 功能还没有完全被理解。CD56或神经细胞黏附分子(NCAM)最早是在 前体细胞，其中它可以与其配体(例如，NCAM， 纤维连接蛋白结构域，硫酸盐蛋白多糖)，并调节神经元的生存、迁移和发育。 有趣的是，异种小鼠发育支持基质(如EL08.1D2和OP9)通常 用于从CD34+祖细胞和造血部位的基质细胞体外分化NK细胞(例如， 骨髓间充质干细胞)与不支持发育的基质相比，表达NCAM。此外， Mace实验室发表的研究表明，抑制CD56(NCAM)介导的相互作用 NK细胞和基质细胞受封闭抗体干扰体外发育导致NK细胞减少 NK细胞数量和迁移。我假设NCAM在支持发育的基质细胞和 NK细胞表面CD56介导NK细胞所需的基质细胞与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的功能角色 在支持发育的基质上的表达。我的初步数据显示，Ncam1的使用 敲除小鼠EL08.1D2间质细胞体外分化NK细胞导致NK细胞减少 发育中间的生存、增殖和迁移。我将确定NCAM，on 支持发育的基质，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CD56在人类NK细胞发育和发育中的要求 迁移。Mace实验室的研究表明，通过封闭抗体抑制NK细胞上的CD56会导致 细胞迁移的减少和肌动蛋白细胞骨架的失调。我将删除人NK细胞中的CD56 并通过转录和表型分析跟踪其体外发育。在一起我们的 研究将揭示一种以前未知的CD56介导的相互作用和信号通路 人类NK细胞在发育过程中增殖、存活和迁移所必需的。

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.