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: 9758122
• 负责人: Emily Chiu
• 金额: $ 4.07万
• 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/9758122
• 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 急性髓系白血病(AML)是一种极具挑战性的疾病。目前唯一已知的“解药”是 造血干细胞移植(HSCT)，需要在移植前进行调理。这个 受急性髓系白血病影响的人群不能很好地耐受清髓性调节，因此降低强度的调节 而不是使用。然而，在接受强度降低的患者中，复发率高得令人无法接受。 条件反射。免疫疗法是治疗所有类型癌症的一个很有前途的新领域，我们的目标是改进 急性髓系白血病患者的免疫治疗。靶向NK细胞的好处在于其固有的肿瘤监视 才能。获得性NK细胞是NK的一种亚型，在巨细胞病毒(CMV)激活后被扩增 表现出增强杀伤力和细胞因子产生的细胞。适应性NK细胞被认为是适应性的，因为 它们的记忆样表型，在再次暴露后表现出增强的激活。适应性NK细胞也是 与HSCT后较低的复发率相关。适应性NK细胞很容易被鉴定为CD57+NKG2C+。这个 NKG2C在适应性NK细胞中的作用尚未完全明确。NKG2C与人类白细胞抗原-E结合 经典的主要组织相容性复合体(MHC)I类分子，具有激活NK细胞的多肽特异性。 NKG2C+细胞还下调NKG2C家族成员NKG2a的表达，NKG2a也通过多肽与人类白细胞抗原E结合 特异性，但抑制NK细胞。我们的建议旨在开发NKG2C用于肿瘤的适应性NK细胞 免疫疗法。在目标1中，我们研究了NKG2C在适应性NK细胞中的作用。我们将使用一种细胞系， K562，缺乏MHC-I类分子，由于缺乏抑制作用而导致强大的NK细胞激活。我们会 操纵这些细胞表达人类白细胞抗原E(K562-E)，并与这些细胞一起确定是否存在信号转导 是通过适应性NK细胞特有的NKG2C发挥作用，还是通过NKG2A表达减少发挥作用 随后减少适应性NK细胞的抑制，导致其功能增强。我们可以利用 外周血NK细胞研究NKG2C对适应性NK细胞的作用，但适应性NK细胞的数量 人类的细胞是多变的。在目标2中，我们将使用诱导多能干细胞(IPSC)来源的NK细胞(INK)和 转导NKG2C杀伤肿瘤细胞。我们将使用三特异性杀伤分子来特异性地靶向肿瘤细胞 激活墨水上的NKG2C，将它们与肿瘤靶点连接起来，同时刺激NKG2C和 IL15受体。AIM 2创造了一种特定的转化疗法，可以扩展到临床试验中。总体而言，这 该项目旨在开发NKG2C作为免疫治疗的功能性受体。