Embryonic natural killer cell development and function

项目总结/摘要 我们研究的总体目标是了解胚胎天然的起源、发育和功能， 杀伤（NK）细胞。NK细胞是先天性免疫系统淋巴细胞，目前是细胞免疫的目标群体。 基于过继免疫疗法。然而，NK细胞疗法已被证明是有效的，只有在一个。 这可能部分是由于在不同的患者中发现的NK细胞亚群的高度异质性。 捐助者。相比之下，从人多能干细胞（hPSC）产生NK细胞是一个特别有利的方法。 令人兴奋的替代方案，因为它将消除供体来源之间的遗传变异性，确保高度可重复性 产生抗肿瘤NK细胞。虽然成人的NK细胞来源于造血干细胞（HSC）， 尽管NK细胞是由淋巴样祖细胞衍生的，但对NK细胞的发育起源仍然知之甚少。我们有 证明了HSC非依赖性造血在早期鼠胚胎中以多波发生， 包括多能“红骨髓祖细胞”（EMP）。同样，我们已经开发了一种体外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，HSC独立， “胚外”红骨髓祖细胞可以产生NK细胞，其在形态上和 在功能上不同于“胚胎内”hPSC衍生的定形祖细胞或新生供体。重点 这项提案的目的是汇集小鼠发育生物学和hPSC指导的专业知识， 分化，作为互补模型系统，以鉴定胚胎NK细胞的起源，及其 平移势我们假设这些胎儿NK细胞是NK细胞的临床相关来源， 强大的抗肿瘤能力。我们将在三个具体目标中检验这一假设。在目标1中，我们 表征小鼠胚胎和hPSC衍生的NK细胞的个体发生起源，建立它们的谱系 从多潜能骨髓祖细胞和转录调控程序的规范。在目标2中， 将建立胚胎NK细胞对成人淋巴细胞库的贡献， 表征HSC非依赖性NK细胞的过继转移抗肿瘤潜力。圆满完成 这些研究将为我们提供更全面的了解哺乳动物造血 发展这对于我们理解先天性发育是至关重要的。 免疫系统和NK细胞的体外产生，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.