Exploiting NKG2C on adaptive NK cells for Immunotherapy

利用适应性 NK 细胞上的 NKG2C 进行免疫治疗

• 批准号: 9920000
• 负责人: Emily Chiu
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920000
• 关键词: Acute Myelocytic Leukemia; Adaptor Signaling Protein; Address; Adult; Affect; Age; Binding; C-Type Lectins; CD8-Positive T-Lymphocytes; Cell Line; Cells; Cessation of life; Clinical; Clinical Trials; Cytolysis; Cytomegalovirus; Development; Disease; Disease-Free Survival; Equilibrium; Exclusion; Exhibits; FCGR3B gene; Family; Family member; Frequencies; HLA Antigens; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hepatitis B e Antigens; Human; Immunotherapy; In Vitro; Individual; Ink; Interleukin-15; K-562; K562 Cells; KLRD1 gene; Knowledge; Ligation; Link; Major Histocompatibility Complex; Malignant - descriptor; Measurable; Mediating; Membrane; Memory; NK Cell Activation; Natural Killer Cells; Patients; Pattern; Peptides; Phenotype; Physiological; Play; Population; Production; Regimen; Relapse; Role; Signal Transduction; Specificity; Stem cell transplant; Surface; T-Lymphocyte Subsets; TYROBP gene; Technology; Testing; Therapeutic; Transcriptional Regulation; Transplantation; Tumor Burden; Tyrosine; Viral; arm; base; cancer therapy; cancer type; cell transformation; chemoradiation; chemotherapy; conditioning; curative treatments; cytokine; cytotoxicity; dimer; experimental study; extracellular; high risk; immunological synapse; improved; in vivo; induced pluripotent stem cell; member; mortality; mouse model; myeloid leukemia cell; neoplasm immunotherapy; neoplastic cell; neutrophil; new therapeutic target; novel; older patient; peripheral blood; prevent; protein expression; receptor; relapse risk; response; seropositive; trispecific killer engager; tumor

Project Summary/Abstract Acute myeloid leukemia (AML) is a challenging disease to cure. The only known “cure” currently is hematopoietic stem cell transplantation (HSCT) which requires conditioning prior to transplantation. The population that AML affects does not tolerate myeloablative conditioning well so a reduced intensity conditioning is used instead. However, the relapse rate is unacceptably high in patients that receive reduced intensity conditioning. Immunotherapy is a promising new field for treatment of all cancer types and we aim to improve immunotherapy for AML patients. The benefits of targeting NK cells lie within its inherent tumor surveillance ability. Adaptive NK cells, which are expanded after cytomegalovirus (CMV) reactivation, are a subtype of NK cells that display enhanced killing and cytokine production. Adaptive NK cells are considered adaptive because of their memory-like phenotype, showing enhanced activation following re-exposure. Adaptive NK cells are also associated with lower relapse following HSCT. Adaptive NK cells are easily identified as CD57+NKG2C+. The function of NKG2C has not been thoroughly defined on adaptive NK cells. NKG2C binds to HLA-E, a non- classical major histocompatibility complex (MHC) class I molecule, with peptide specificity to activate NK cells. NKG2C+ cells also downregulate NKG2A, a family member of NKG2C that also binds to HLA-E with peptide specificity but inhibits NK cells. Our proposal aims to exploit NKG2C on adaptive NK cells for tumor immunotherapy. In Aim 1, we look at the role of NKG2C specifically on adaptive NK cells. We will use a cell line, K562, that lack MHC class I molecules resulting in potent NK cell activation due to lack of inhibition. We will manipulate these cells to express HLA-E (K562-E) and with these cells to determine whether there is a signaling function through NKG2C that is unique to adaptive NK cells or whether it is the decrease in NKG2A expression and subsequent reduction of inhibition in adaptive NK cells that leads to their enhanced function. We can use peripheral blood NK cells to study the function of NKG2C on adaptive NK cells, but the number of adaptive NK cells in humans is variable. In Aim 2, we will use induced pluripotent stem cell (iPSC) derived NK cells (iNK) with transduced NKG2C to kill tumor cells. We will specifically target tumor cells using a Trispecific Killer Engager that activates NKG2C on the iNK, linking them with tumor target while simultaneously stimulating NKG2C and IL15 receptor. Aim 2 creates a specific translational therapy that can be scaled for clinical trials. Overall, this project aims to exploit NKG2C as a functional receptor for immunotherapy.

项目摘要/摘要 急性髓样白血病（AML）是治愈的挑战疾病。目前唯一已知的“治愈”是 造血干细胞移植（HSCT），需要在移植之前进行调节。这 AML影响的人群不能很好地忍受髓质调节，因此强度降低 改用。但是，在强度降低的患者中，继电器速率不可接受 调理。免疫疗法是一种治疗所有癌症类型的新领域，我们旨在改善 AML患者的免疫疗法。靶向NK细胞的好处在于其继承的肿瘤监测 能力。自适应NK细胞在巨细胞病毒（CMV）重新激活后扩展，是NK的亚型 表现出增强的杀伤和细胞因子产生的细胞。自适应NK细胞被认为是自适应的，因为 它们的记忆状表型，显示重新暴露后的激活增强。自适应NK细胞也是 与HSCT后较低的继电器相关。自适应NK细胞很容易识别为CD57+NKG2C+。这 NKG2C的功能尚未在自适应NK细胞上彻底定义。 NKG2C与HLA-E结合，一种非 - 经典的主要组织相容性复合物（MHC）I类分子，具有肽特异性以激活NK细胞。 NKG2C+细胞还下调NKG2A，NKG2A是NKG2C的家庭成员，它也与胡椒结合HLA-E 特异性但抑制NK细胞。我们的建议旨在利用自适应NK细胞上的NKG2C进行肿瘤 免疫疗法。在AIM 1中，我们研究了NKG2C在自适应NK细胞上的作用。我们将使用一个单元线， K562，缺乏MHC I类分子，导致由于缺乏抑制作用而导致潜在的NK细胞活化。我们将 操纵这些细胞以表达HLA-E（K562-E），并使用这些细胞来确定是否存在信号 通过自适应NK细胞独有的NKG2C函数，还是NKG2A表达的降低 随后减少自适应NK细胞的抑制作用，从而导致其功能增强。我们可以使用 周围血液NK细胞研究NKG2C在自适应NK细胞上的功能，但是自适应NK的数量 人类中的细胞是可变的。在AIM 2中，我们将使用诱导的多能干细胞（IPSC）衍生的NK细胞（墨水） 转导NKG2C杀死肿瘤细胞。我们将使用三特异性杀手参与者专门针对肿瘤细胞 激活墨水上的NKG2C，将它们与肿瘤靶标连接在一起，同时刺激NKG2C和 IL15受体。 AIM 2创建了一种特定的翻译疗法，可以缩放以进行临床试验。总体而言，这 项目旨在利用NKG2C作为免疫疗法的功能接收器。