Control of Cytotoxic Lymphocytes by Polymorphic KIR3DL3

多态性 KIR3DL3 控制细胞毒性淋巴细胞

• 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
• 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

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天然细胞毒性淋巴细胞，包括自然杀伤（NK）细胞，是免疫系统的重要组成部分。 3感染和恶性肿瘤的监测。它们的效应子功能通过多种 淋巴细胞与靶细胞之间的免疫突触上有4对配体-受体。在这方面，杀手细胞 5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杀伤， 6鼓励杀死不表达其配体的不健康细胞。个体和群体在两个方面都有差异， 7存在的KIR基因的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， 9.造血细胞移植和繁殖的成功。KIR 3DL 3的独特之处在于它是唯一一款 10在多个灵长类物种中是保守的，并在每个人类个体中观察到。 尽管KIR 3DL 3的普遍存在强调了它在人类生存中的必要性，但我们缺乏对KIR 3DL 3的基本了解。 12其生物学作用，包括信号传导能力、配体相互作用、对下游效应子功能的影响 13和它可以表达的组织。虽然我们的总体目标是建立功能， 14伴侣和组织分布，我们还将探索其用于免疫治疗的效用。基本 为了证明后者，我们已经鉴定了KIR 3DL 3的配体，其可以用于帮助NK细胞特异性地抑制NK细胞的增殖。 16检测和消除感染或癌细胞。该配体与其他标记物密切相关， 17例已成功用于免疫检查点抑制治疗。在目标1中，我们将描述KIR 3DL 3配体 18并定义这种相互作用在NK细胞生物学中的作用。我们将测量相互作用的动力学性质， 19产生复合物的晶体结构并进行受体后效应子功能的测定 20例结扎。在目标2中，我们将使用生物化学方法来表征KIR 3DL 3细胞内的需求。 21信号传导和定义相关的途径。其他KIR在其结构域中含有两个特定的氨基酸基序。 22介导抑制信号的胞质尾区。KIR 3DL 3只包含其中一个基序，我们的进化 23分析表明，受体聚集是使它们靠近以进行信号传导所必需的。 24在我们的初步分析中，我们在一组组织中发现了KIR 3DL 3的表达，这些与 25我们鉴定了要表达的配体的组织。为了研究组织驻留在 26细胞毒性细胞通过KIR 3DL 3与特定组织细胞相互作用，目的3，我们将在体内测定KIR 3DL 3的表达。 27受体和配体的表达谱。我们还将设计一种嵌合的KIR 3DL 3，它可以指导NK细胞， 28杀死，而不是保留表达配体的不健康细胞。KIR 3DL 3是高度多态的，并且是最重要的基因之一。 29个杂合子人类基因。因此，在我们所有的目标，我们将评估多态性的影响， KIR 3DL 3的30个属性和功能。