Embryonic natural killer cell development and function

项目摘要/摘要 我们研究的总体目标是了解胚胎自然的起源、发展和功能 杀伤(NK)细胞。NK细胞是先天免疫系统的淋巴细胞，目前是细胞的靶群。 以过继免疫疗法为基础。然而，NK细胞疗法已被证明只在 患者的亚群，可能部分是由于在 捐赠者。相比之下，从人类多能干细胞(HPSCs)产生NK细胞是一个特别重要的问题 令人兴奋的替代方案，因为它将消除捐赠者来源之间的遗传变异，确保高度可重复性 抗肿瘤NK细胞的产生。而成人的NK细胞来自造血干细胞(HSC)- 作为原始淋巴祖细胞，NK细胞的发育起源目前仍知之甚少。我们有 证明在小鼠早期胚胎中，HSC非依赖的造血发生在多波中， 包括多潜能的“红系祖细胞”(EMP)。同样，我们已经开发出一种体外hPSC 阶段特异性分化方法概括了胚胎造血的许多方面 发展。结合这些系统，我们的初步平行研究表明，不依赖于HSC， “胚胎外”的红系祖细胞可以产生NK细胞，在形态上和 在功能上不同于“胚胎内”hPSC衍生的最终祖细胞，或新生儿捐赠者。焦点 这项建议的目的是将小鼠发育生物学和hPSC指导的专业知识结合起来 分化作为互补模型系统用于鉴定胚胎NK细胞的来源(S)及其 翻译潜力。我们假设这些胎儿NK细胞是临床上相关的NK细胞来源， 强大的抗肿瘤能力。我们将通过三个具体目标来检验这一假设。在目标1中，我们将 鉴定小鼠胚胎和hPSC来源的NK细胞的个体起源，建立其谱系 来自多潜能髓系祖细胞的规范和转录调控程序。在目标2中，我们 将建立胚胎NK细胞对成人淋巴细胞库的贡献，并在功能上 鉴定HSC非依赖性NK细胞过继转移抗肿瘤的潜能。圆满完成 这些研究将为我们提供对哺乳动物造血的更全面的了解 发展。这对于我们理解人的先天发展是至关重要的。 免疫系统和NK细胞的体外生成，用于广泛的再生医学应用， 包括抗癌治疗。

PROJECT SUMMARY / ABSTRACT The overall goal of our research is to understand the origin, development and function of embryonic Natural Killer (NK) cells. NK cells are innate immune system lymphocytes and a current target population for cell- based adoptive immunotherapies. However, NK cell therapy has been demonstrated to be effective in only a subset of patients, possibly due in part to the highly heterogeneous nature of NK cell subsets found across donors. In contrast, the generation of NK cells from human pluripotent stem cells (hPSCs) is a particularly exciting alternative, as it would eliminate genetic variability across donor sources, ensuring highly reproducible generation of anti-tumor NK cells. While NK cells in the adult are derived from hematopoietic stem cell (HSC)- derived lymphoid progenitors, the developmental origin of NK cells remains poorly understood. We have demonstrated that HSC-independent hematopoiesis occurs in multiple waves in the early murine embryo, including multipotential “erythro-myeloid progenitors” (EMP). Similarly, we have developed an in vitro hPSC stage-specific differentiation approach that recapitulates many aspects of embryonic hematopoietic development. Combining these systems, our preliminary parallel studies indicate that HSC-independent, “extra-embryonic” erythro-myeloid progenitors can generate NK cells, which are morphologically and functionally distinct from “intra-embryonic” hPSC-derived definitive progenitors, or neonatal donors. The focus of this proposal is to bring together expertise in mouse developmental biology and hPSC directed differentiation, as complementary model systems, to identify the origin(s) of embryonic NK cells, and their translational potential. We hypothesize that these fetal NK cells are a clinically-relevant source of NK cells, with robust antitumor capacity. We will test this hypothesis across three Specific Aims. In Aim 1, we will characterize the ontogenic origins of murine embryonic and hPSC-derived NK cells, establishing their lineage specification from a multipotential myeloid progenitor and the transcriptional regulatory programs. In Aim 2, we will establish the contribution of embryonic NK cells to the adult lymphocyte repertoire, and functionally characterize the adoptive transfer antitumor potential of HSC-independent NK cells. The successful completion of these studies will provide us with a more comprehensive understanding of mammalian hematopoietic development. This is of fundamental importance to our understanding of the development of the innate immune system and the in vitro generation of NK cells for a wide array of regenerative medicine applications, including anti-cancer therapy.