Control of Cytotoxic Lymphocytes by Polymorphic KIR3DL3

多态性 KIR3DL3 控制细胞毒性淋巴细胞

基本信息

批准号：
10469872
负责人：
Paul John Norman
金额：
$ 36万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-07 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10469872
关键词：
Alleles Allergic Amino Acid Motifs Autoimmune Diseases Autoimmunity Binding Biochemical Biological Biological Assay Biological Process Biology Cancerous Cell Line Cell surface Cells Cellular Assay Cellular biology Complex Crystallization Crystallography Cytoplasmic Tail Data Decidua Dependence Detection Dimerization Disease Drug or chemical Tissue Distribution Duodenum Effector Cell Engineering Family Gene Family Genes Genetic Polymorphism Goals Health Human Human Herpesvirus 4 ITIM Immune Immune Targeting Immunity Immunoglobulins Immunologic Surveillance Immunotherapeutic agent Immunotherapy In Vitro Individual Infection Infection Control Inflammatory KLRA1 gene Killer Cells Kinetics Ligands Ligation Lung Lymphocyte Malignant Neoplasms Measures Mediating NK cell receptor NKB1 Natural Killer Cells Natural Selections Pathway interactions Phosphoric Monoester Hydrolases Placentation Population Predisposition Primates Property RNA Interference Receptor Aggregation Receptor Gene Receptor Signaling Regulation Reporter Reproduction Role Signal Transduction Specificity Stains Stimulus Structure Surface Plasmon Resonance Testing Therapeutic Tissue Sample Tissues Variant cell killing checkpoint inhibition cytokine cytotoxic cytotoxicity dimer genetic variant hematopoietic cell transplantation immune checkpoint immunological synapse in vivo mutant receptor recruit reproductive system disorder response single-cell RNA sequencing success

项目摘要

1 Summary 2 Innate cytotoxic lymphocytes, including natural killer (NK) cells, are essential components of immune 3 surveillance for infection and malignancy. Their effector functions are modulated through interaction of multiple 4 ligand-receptor pairs at the immune synapse between lymphocyte and target cell. In this regard, killer cell 5 immunoglobulin-like receptors (KIR) can suppress killing of any healthy cells that express their ligand and 6 encourage killing of unhealthy cells that do not express their ligand. Individuals and populations vary both in the 7 number of KIR genes present and the specific alleles of those genes. This extreme polymorphism of the KIR 8 gene family is implicated in susceptibility to infectious, allergic, inflammatory, and autoimmune diseases, and in 9 the success of hematopoietic cell transplantation and reproduction. KIR3DL3 is unique in being the only KIR that 10 is conserved through multiple primate species and observed in every human individual. 11 Whereas the ubiquity of KIR3DL3 underlines its necessity in human survival, we lack a basic understanding of 12 its biological role, including signalling capability, ligand interactions, influence on downstream effector functions 13 and the tissues where it can be expressed. While our overarching goal is thus to establish the function, binding 14 partner and tissue distribution of KIR3DL3, we will also explore its utility for immunotherapy. Substantially 15 justifying the latter, we have identified a ligand for KIR3DL3 that could be utilized to aid NK cells in specific 16 detection and elimination of infected or cancerous cells. The ligand is closely related to other markers that have 17 been successfully used for immune checkpoint inhibition therapy. In Aim 1, we will describe the KIR3DL3 ligand 18 and define the role of this interaction in NK cell biology. We will measure the kinetic properties of the interaction, 19 generate a crystal structure of the complex and perform assays of the effector functions following receptor 20 ligation. In Aim 2, we will use biochemical approaches to characterize the requirements for KIR3DL3 intracellular 21 signalling and define the associated pathways. Other KIR contain two specific amino acid motifs in the 22 cytoplasmic tail that mediate inhibitory signals. KIR3DL3 contains only one of these motifs, and our evolutionary 23 analysis suggests that receptor aggregation is necessary to bring them into proximity for signalling. 24 In our preliminary analysis we identified KIR3DL3 expression in a subset of tissues, and these are the same as 25 the tissues where we identified the ligand to be expressed. To investigate the hypothesis that tissue resident 26 cytotoxic cells interact with specific tissue cells through KIR3DL3, in Aim 3, we will determine the in vivo 27 expression profile of receptor and ligand. We will also engineer a chimeric KIR3DL3 that can direct NK cells to 28 kill, rather than spare unhealthy cells that express the ligand. KIR3DL3 is highly polymorphic and one of the most 29 heterozygous human genes. Thus, throughout all our aims, we will assess the impact of polymorphism on the 30 properties and functions of KIR3DL3.

1 总结 2 先天细胞毒性淋巴细胞，包括自然杀伤 (NK) 细胞，是免疫的重要组成部分 3 监测感染和恶性肿瘤。它们的效应器功能通过多个相互作用来调节淋巴细胞和靶细胞之间的免疫突触处有 4 个配体-受体对。对此，杀伤细胞 5 种免疫球蛋白样受体 (KIR) 可以抑制任何表达其配体的健康细胞的死亡 6 鼓励杀死不表达其配体的不健康细胞。个体和人群在以下方面都有所不同 7 存在的 KIR 基因数量以及这些基因的特定等位基因。 KIR 的这种极端多态性 8 基因家族与感染性、过敏性、炎症性和自身免疫性疾病的易感性有关，并且与 9.造血细胞移植和繁殖成功。 KIR3DL3 的独特之处在于它是唯一一个 10 在多种灵长类动物中是保守的，并且在每个人类个体中都观察到。 11 尽管 KIR3DL3 的普遍存在强调了它在人类生存中的必要性，但我们对 12 其生物学作用，包括信号传导能力、配体相互作用、对下游效应功能的影响 13 及其可以表达的组织。虽然我们的首要目标是建立功能，但绑定 14 KIR3DL3的伙伴和组织分布，我们还将探索其在免疫治疗中的效用。实质上 15 为了证明后者的合理性，我们已经鉴定出 KIR3DL3 的配体，可用于帮助 NK 细胞进行特定的治疗。 16 检测并消除感染细胞或癌细胞。该配体与其他标记物密切相关 17已成功用于免疫检查点抑制治疗。在目标 1 中，我们将描述 KIR3DL3 配体 18 并定义了这种相互作用在 NK 细胞生物学中的作用。我们将测量相互作用的动力学特性， 19 生成复合物的晶体结构并进行受体后效应子功能的测定 20结扎。在目标 2 中，我们将使用生化方法来表征 KIR3DL3 细胞内的要求 21 信号传导并定义相关途径。其他 KIR 中含有两个特定的氨基酸基序 22 介导抑制信号的细胞质尾部。 KIR3DL3 仅包含这些基序之一，而我们的进化 23 分析表明，受体聚集对于使它们接近信号传导是必要的。 24 在我们的初步分析中，我们确定了一部分组织中的 KIR3DL3 表达，这些表达与 25 我们鉴定出要表达的配体的组织。调查组织驻留的假设 26种细胞毒性细胞通过KIR3DL3与特定组织细胞相互作用，在目标3中，我们将确定体内图27受体和配体的表达谱。我们还将设计一个嵌合 KIR3DL3，它可以引导 NK 细胞 28 杀死而不是保留表达配体的不健康细胞。 KIR3DL3 具有高度多态性，是最常见的多态性之一。 29个杂合的人类基因。因此，在我们所有的目标中，我们将评估多态性对 KIR3DL3的30个属性和功能。