The role of Notch signaling in human natural killer cell functional maturation

项目摘要/摘要 我们在这个项目中的长期目标是阐明人类自然杀伤细胞发育的机制 目的：探讨NK细胞生物学过程在肿瘤治疗中的应用。自然杀伤(NK)细胞识别 恶性细胞表面缺乏自身MHC分子，因此以穿孔素为靶点 细胞毒性。NK细胞在对恶性肿瘤的先天免疫反应中扮演着重要的角色，事实上 癌症，如急性髓系白血病(AML)，已被证明损害了NK细胞的发育和 功能。因此，了解目前尚不清楚的调节NK细胞成熟的机制 正常和疾病环境将改善癌症治疗的转化研究努力。我们正在进行的工作 研究次级淋巴组织(SLT；包括扁桃体)中的NK细胞发育途径 和淋巴结)。在我们目前的研究中，我们观察到Notch信号通路的激活导致 从非功能表型到功能成熟的NK细胞发育过程中的关键转变。此外， 我们在新鲜的成熟NK细胞中检测到了Notch转录活性区域的组成性表达 从扁桃体中分离出来，从而支持Notch在调节功能表型中的生理学重要性 成熟的自然杀伤细胞。我们假设SLT微环境中的Notch信号调节 从未成熟的NK细胞发育到具有功能的NK细胞。我们进一步假设，通过 我们观察到在AML中激活的芳烃受体(AHR)改变Notch信号以抑制NK细胞 发展。因此，我们的目标是1)确定Notch在SLT中被激活以促进 2)确定AHR如何调节Notch，以对抗NK细胞的成熟。 为了验证这一假说，我们将进行体外培养研究和体内领养移植实验。 从人扁桃体中分离出NK和其他SLT驻留细胞。我们将确定激活的直接基因靶点 以确定Notch如何调节NK细胞功能的关键分子的转录。 最后，我们将确定AHR如何调节NK细胞中的Notch信号。这个项目的总体意义 将解决SLT中关于调节人类NK细胞成熟的相关知识空白 微环境。该项目代表了癌症免疫学的一种新颖和创新的方法，因为它 结合了对重要信号通路的机制研究，并试图确定癌症是如何 例如急性髓系白血病可以逃避人类的免疫系统。成功完成这些目标将提高我们的 了解自然杀伤细胞发育的正常过程，目标是将这些发现应用于 基于免疫的癌症新疗法。

PROJECT SUMMARY/ABSTRACT Our long-term goal in this project is to elucidate the mechanisms of human natural killer cell development in order to apply the processes of NK cell biology in the treatment of cancer. Natural killer (NK) cells recognize malignant cells lacking self-MHC molecules on their surface, thus targeting them for perforin-mediated cytotoxicity. NK cells comprise an important role in the innate immune response to malignancy, and indeed cancers such as acute myeloid leukemia (AML) have been shown to have impaired NK cell development and function. Therefore, understanding the yet unknown mechanisms regulating NK cell maturation in both the normal and disease settings will improve translational research efforts for cancer therapy. Our ongoing work investigates NK cell developmental pathways occurring in secondary lymphoid tissues (SLTs; including tonsils and lymph nodes). In our current studies, we observed that activation of the Notch signaling pathway results in a key transition during development from a non-functional phenotype to a functionally mature NK cell. Moreover, we detected constitutive expression of the transcriptionally-active domain of Notch in mature NK cells freshly isolated from tonsils, thus supporting the physiologic importance of Notch in regulating the functional phenotype of mature NK cells. We hypothesize that Notch signaling in the SLT microenvironment regulates the developmental transition from immature to functional NK cells. We further hypothesize that signaling through the aryl hydrocarbon receptor (AHR), which we observe is activated in AML, alters Notch signaling to inhibit NK cell development. Thus our aims are 1) to determine the mechanism by which Notch is activated in SLTs to promote NK cell functional maturation; and 2) to determine how AHR modulates Notch to antagonize NK cell maturation. To test this hypothesis, we will perform in vitro culture studies and in vivo adoptive transfer experiments using NK and other SLT-resident cells isolated from human tonsils. We will identify the direct gene targets of activated Notch in NK cells to determine how Notch regulates the transcription of critical molecules for NK cell function. Finally, we will identify how AHR modulates Notch signaling in NK cells. The overall significance of this project will address a relevant gap in knowledge regarding the regulation of human NK cell maturation in the SLT microenvironment. This project represents a novel and innovative approach to cancer immunology because it combines both mechanistic studies of an important signaling pathway, and also seeks to identify how cancers such as AML can evade the human immune system. Successful completion of these aims will improve our understanding of the normal processes of NK cell development with the goal of applying these discoveries toward novel immune-based therapies for cancer.