Delineating novel functions of Semaphorin 3E in regulating natural killer cell biology

描述 Semaphorin 3E 在调节自然杀伤细胞生物学中的新功能

• 批准号: RGPIN-2022-04504
• 负责人: Kung, SamKP
• 金额: $ 2.33万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=757195
• 关键词: Delineating; novel; functions; Semaphorin; 3E

Natural killer (NK) cells are important immune cells that regulate innate immunity. NK cells are activated by receptor recognition of target cells, cytokines or dendritic cells (DC) in their microenvironments. How specific factor(s) found in their local microenvironments regulate NK cell biology remains to be defined. My NSERC-funded research program seeks to achieve a molecular understanding of how factors found in microenvironments regulate NK-cell development, recruitment and/or NK cell effector functions. One of the key findings I have obtained in the last funding cycle is the novel role of Semaphorin 3E (Sema3E), a protein originally identified for its roles in neurodevelopment, in regulating NK-cell migration in the NK-DC crosstalk. The objective of my current NSERC-DG proposal is to delineate novel functions of the Sema3E/PlexinD1 pathway in regulating various aspects of NK cell biology under the experimental microenvironments we defined by cytokines and NK-DC crosstalk. Global Hypothesis: The Sema3E/Plexin D1 axis is a novel signaling pathway that regulates NK cell maturation, effector functions (cytotoxicity, cytokine production) and/or migratory properties. Specific Objectives: 1. To examine the maturation program of NK cells in Sema3E deficient mice. I will quantify immature and mature NK cell subsets in spleen and bone marrow. I will examine proliferation, apoptosis, exhaustion phenotypes and cell cycle in the ex vivo NK and activated NK cells. 2. To delineate molecular mechanism underlying Sema3E regulation of NK cell functions. I will determine whether Sema3E deficiency impacts migratory properties of NK cells under different defined microenvironments in vitro. I will elucidate signaling networks that are impacted by Sema3E deficiency, and how they affect NK cell effector functions. 3. To determine whether Sema3E deficiency directly regulate NK cell development and functions via the cognate Sema3E receptor (Plexin D1) on NK cells. I will use recombinant Sema3E in in vitro NK cultures to examine whether direct activation of the PlexinD1 receptor on NK cells modulates NK cell functions. I will create a novel NK specific Plexin D1 KO mouse model to examine NK cell development and functions in our established assays. Significance: Discoveries from this proposed work will advance our understanding of how this novel Sema3E/PlexinD1 pathway regulates NK cell lymphocyte biology in both sexes, and provide new insights into how this and other novel pathways operate in specific microenvironments. The novelty of the research projects covered in this cycle, the Kung laboratory that supports equity, diversity and inclusion in research, the cutting edge technologies and the experimental approaches involved will train and prepare my HQP well to lead research projects in either academic or private sectors.

自然杀伤（NK）细胞是调节先天免疫的重要免疫细胞。NK细胞通过其微环境中的靶细胞、细胞因子或树突状细胞（DC）的受体识别而被激活。在其局部微环境中发现的特定因子如何调节NK细胞生物学仍有待确定。我的NSERC资助的研究计划旨在实现微环境中发现的因素如何调节NK细胞发育，募集和/或NK细胞效应功能的分子理解。我在上一个资助周期中获得的关键发现之一是Semaphorin 3E（Sema 3E）的新作用，Sema 3E是一种最初因其在神经发育中的作用而被鉴定的蛋白质，在NK-DC串扰中调节NK细胞迁移。我目前的NSERC-DG建议的目的是描绘新的功能Sema 3E/丛蛋白D1途径在调节NK细胞生物学的各个方面下，我们定义的细胞因子和NK-DC串扰的实验微环境。 全局假设：Sema 3E/丛状蛋白D1轴是一种新的信号传导途径，可调节NK细胞成熟、效应子功能（细胞毒性、细胞因子产生）和/或迁移特性。具体目标：1。研究Sema 3E缺陷小鼠NK细胞的成熟程序。我将量化脾脏和骨髓中未成熟和成熟的NK细胞亚群。我将检查体外NK细胞和活化NK细胞的增殖、凋亡、耗竭表型和细胞周期。2.阐明Sema 3E调节NK细胞功能的分子机制。我将确定Sema 3E缺陷是否影响NK细胞在不同定义的体外微环境下的迁移特性。我将阐明受Sema 3E缺陷影响的信号网络，以及它们如何影响NK细胞效应功能。3.确定Sema 3E缺陷是否通过NK细胞上的同源Sema 3E受体（丛状蛋白D1）直接调节NK细胞发育和功能。我将在体外NK培养中使用重组Sema 3E来检查NK细胞上的丛蛋白D1受体的直接激活是否调节NK细胞功能。我将创建一个新的NK特异性丛状蛋白D1 KO小鼠模型，以检查NK细胞的发育和功能，在我们建立的测定。重要性：这项工作的发现将促进我们对这种新型Sema 3E/PlexinD 1通路如何调节两性NK细胞淋巴细胞生物学的理解，并为这种和其他新型通路如何在特定微环境中运作提供新的见解。本周期所涵盖的研究项目的新奇，支持研究公平，多样性和包容性的Kung实验室，所涉及的尖端技术和实验方法将训练和准备我的HQP，以领导学术或私营部门的研究项目。