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.

自然杀伤 (NK) 细胞是人类淋巴细胞的主要谱系，在先天性和适应性方面具有重要功能对血液癌症的免疫和适应性免疫是由多种介导的。高度多态性 HLA-A、-B 和 -C 分子与同样多态性杀伤性免疫球蛋白的相互作用它们通过 HLA-E 与 CD94:NKG2 的保守相互作用得到补充。编码KIR受体和HLA配体的基因由此独立地分离。在家庭和群体中产生独特且多样化的基因型。功能上不同的 NK 细胞库，这是一种对 HCT 结果产生深远影响的自然变异。迄今为止，还没有一个合适的体内异种模型来研究 NK 细胞的发育和通过 HCT 进行教育或验证人类 NK 细胞的过继转移。研究人类 NK 细胞植入和功能的两大障碍：(1) 人类 NK 细胞重建和生存依赖于共同的  链细胞因子 IL-2、IL-7 和 IL-15，它们之间不存在交叉反应 (2) 人类 NK 细胞功能由抑制性受体教育、调节和维持我们现在已经掌握了与 MHC I 类分子的结合，这些分子在物种之间也不会发生交叉反应。利用尖端腺相关病毒 (AAV) 克服细胞因子和 MHC I 类屏障载体介导的基因递送方法转导编码 HLA-A、-B、-C 和 -E 多肽的基因，如以及某些人类细胞因子，对缺乏小鼠来源的 2- 的高度免疫缺陷的 NSG 小鼠微球蛋白，命名为 NSG-B2M-/- 小鼠在特定目标 (SA)1 中，我们将建立人类免疫系统 (HIS)。表达选择的人类细胞因子和 HLA-A*02 和/或 HLA-E 细胞因子 IL-2、IL-3、IL-6、IL-7、IL-的小鼠。 15和GM-CSF将促进人类NK细胞以及人类T细胞、B细胞、骨髓细胞的发育我们将应用 42 重质谱流式细胞术抗体组来仔细绘制淋巴细胞图谱。 HCT 后的 NK 细胞发育、教育和功能将以前所未有的分辨率进行重建。在 20 周的范围内进行纵向研究在 SA2 中，我们将用荧光素酶标记的肿瘤感染 HIS 小鼠。靶细胞，以评估 NK 细胞教育对抗肿瘤活性和 HCT 结果的影响。将通过质谱流式细胞仪比较来自 HIS 小鼠的 NK 细胞的抗肿瘤功能以及通过我们的 HIS 小鼠模型将为 HLA-E 是否可以提供明确的答案。直接教育 CD94:NKG2A+ 人类 NK 细胞以及这种教育如何影响它们的反应能力在 SA2 中，我们还将测量 CMV 感染（和重新激活）对循环肿瘤的体内影响。培养适应性 NK 细胞及其增强的抗肿瘤反应，最终目标是开发。表达人类细胞因子以及复杂的 HLA I 类单倍型的 HIS 小鼠，我们将能够有效地利用 NK 细胞效应器治疗血液癌症。