Harnessing NK cell effector function for immunotherapies

利用 NK 细胞效应功能进行免疫治疗

基本信息

批准号：
9296856
负责人：
Amir Horowitz
金额：
$ 25.43万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-18 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9296856
关键词：
Activities of Daily Living Address Adoptive Transfer Alleles Alpha Cell Antibodies Antigens Autoimmunity B-Lymphocytes Blood CD34 gene CD4 Positive T Lymphocytes CD8B1 gene Cell Differentiation process Cell physiology Cells Chromosomes Clinical Coculture Techniques Complement Complex Cytokine Signaling Cytomegalovirus Infections Cytometry Dependovirus Development Donor Selection Education Effector Cell Engraftment Environment FCGR3B gene Family Gene Delivery Gene Transfer Genes Genotype Goals Granulocyte-Macrophage Colony-Stimulating Factor HLA Class I Genes HLA-A gene HLA-DR1 Antigen Haplotypes Hematologic Neoplasms Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Histocompatibility Antigens Class I Human Immune Immune response Immune system Immunity Immunization Immunogenetics Immunophenotyping Immunotherapy Infection Interleukin-15 Interleukin-2 Interleukin-3 Interleukin-6 Interleukin-7 KLRD1 gene Kinetics Label Ligands Longitudinal Studies Luciferases Lymphocyte MHC Class I Genes Malignant Neoplasms Maps Measures Mediating Modeling Mus Myelogenous Names Natural Immunity Natural Killer Cells Outcome Phenotype Population Predisposition Process Receptor Cell Resolution Sampling Secondary to Study models Survival Rate T-Lymphocyte Therapeutic Time Transplantation Tumor Burden Vaccines Variant Venous Virus Diseases adaptive immunity adeno-associated viral vector base bioluminescence imaging cytokine improved in vivo in vivo Model killer immunoglobulin-like receptor leukemia macrophage mouse model pathogen polypeptide receptor reconstitution reproductive success response treatment strategy tumor

项目摘要

Natural Killer (NK) cells are a major lineage of human lymphocytes with vital functions in innate and adaptive immunity and adaptive immunity to hematologic cancers. NK cell functions are mediated by the diverse interactions of highly polymorphic HLA-A, -B, and -C molecules with equally polymorphic killer immunoglobulin- like receptors (KIR). They are complemented by the conserved interactions of HLA-E with CD94:NKG2 heterodimers. The genes encoding KIR receptors and HLA ligands segregate independently, thereby generating unique and diverse genotypes within families and populations. Phenotypically, this produces functionally distinct NK cell repertoires, a natural variation that has profound effects on the outcome of HCT. To date, there has never been an appropriate in vivo xenogeneic model for studying NK-cell development and education through HCT or for validating the adoptive transfer of human NK cells. Historically, there have been two major barriers to study human NK-cell engraftment and function: (1) human NK-cell reconstitution and survival is dependent upon common -chain cytokines IL-2, IL-7 and IL-15 that are not cross reactive between species; and (2) human NK-cell function is educated, regulated and maintained by inhibitory receptor engagement with MHC class I molecules that are also not cross-reactive between species. We have now overcome the cytokine and MHC class I barrier by utilizing a cutting-edge adeno-associated virus (AAV) vector-mediated gene delivery approach to transduce genes encoding HLA-A, -B, -C and –E polypeptides, as well as certain human cytokines, to highly immunodeficient NSG mice that lack mouse-derived 2- microglobulin, named NSG-B2M-/- mice. In Specific Aim (SA)1 we will establish human immune system (HIS) mice expressing select human cytokines and HLA-A*02 and/or HLA-E. The cytokines IL-2, IL-3, IL-6, IL-7, IL- 15 and GM-CSF will promote development of human NK cells along with human T cells, B cells, myeloid macrophages and DCs. We will apply a 42-plex mass cytometry antibody panel to closely map lymphocyte reconstitution following HCT at an unprecedented resolution. NK cell development, education and function will be studied longitudinally over a range of 20 weeks. In SA2 we will infect HIS mice with luciferase-labeled tumor target cells in order to evaluate the impact of NK cell education on anti-tumor activity and outcome of HCT. We will compare anti-tumor function of NK cells from HIS mice by mass cytometry as well as tumor burden by bioluminescence imaging. Our HIS mouse model will provide a definitive answer on whether HLA-E can directly educate CD94:NKG2A+ human NK cells and how this education impacts their capacity to respond to circulating tumors. In SA2, we will also measure the in vivo effects of CMV infection (and reactivation) on the education of adaptive NK cells and their enhanced anti-tumor response. With the ultimate goal of developing HIS mice expressing human cytokines along with complex HLA class I haplotypes, we will be able to effectively harness NK cell effector for treatment of hematologic cancers.

天然杀手（NK）细胞是人类淋巴细胞的主要谱系，在先天和适应性中具有重要功能对血液癌的免疫和适应性免疫。 NK细胞功能是由多样的高度多态HLA -A，-b和-c分子与同样多态杀手型免疫球蛋白的相互作用像接收器（KIR）。它们是由HLA-E与CD94的配置交互完成的：NKG2 异二聚体。编码KIR受体和HLA配体的基因独立隔离，从而在家庭和人群中产生独特而多样的基因型。从情景上讲，这会产生在功能上不同的NK细胞库中，一种自然变异对HCT的结果具有深远的影响。到日期，从来没有一个适合研究NK细胞开发和通过HCT教育或验证人NK细胞的适应性转移。从历史上看，有研究人类NK细胞植入和功能的两个主要障碍：（1）人类NK细胞重建和生存依赖于常见的链细胞因子IL-2，IL-7和IL-15，它们在物种; （2）抑制接收器对人类NK细胞功能进行教育，调节和维护与物种之间也没有交叉反应的MHC I类分子的互动。我们现在有通过使用尖端腺相关病毒（AAV）来克服细胞因子和MHC I类屏障向量介导的基因输送方法，用于传播编码HLA -A，-b，-c和 - E多肽的基因以及某些人类细胞因子，高度免疫缺陷的NSG小鼠缺乏小鼠衍生的2- 微球蛋白，称为NSG-B2M - / - 小鼠。在特定目标（SA）1中，我们将建立人类免疫系统（他）表达精选人类细胞因子和HLA-A*02和/或HLA-E的小鼠。细胞因子IL-2，IL-3，IL-6，IL-7，IL- 15和GM-CSF将促进人NK细胞以及人T细胞，B细胞，髓样巨噬细胞和DCS。我们将应用42个质量质量细胞仪抗体面板以密切映射淋巴细胞 HCT以空前的决议进行重建。 NK细胞开发，教育和功能将在20周的范围内纵向研究。在SA2中，我们将用荧光素酶标记的肿瘤感染他的老鼠靶细胞以评估NK细胞教育对HCT抗肿瘤活性和结果的影响。我们将通过质量细胞仪以及肿瘤烧伤的NK细胞的抗肿瘤功能生物发光成像。我们的鼠标模型将为HLA-E是否可以提供明确的答案直接教育CD94：NKG2A+人类NK细胞，以及该教育如何影响其应对能力循环肿瘤。在SA2中，我们还将测量CMV感染（和重新激活）对体内影响自适应NK细胞及其抗肿瘤反应的增强。以发展的最终目标他的老鼠表达人类细胞因子以及复杂的HLA I类单倍型，我们将能够有效线束NK细胞效应子治疗血液学癌症。