Delineating the role of calcineurin in modulating the immune response in acute lymphoblastic leukemia

描述钙调神经磷酸酶在调节急性淋巴细胞白血病免疫反应中的作用

• 批准号: 10361198
• 负责人: Alisha Desiree Hunter
• 金额: $ 3.33万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10361198
• 关键词: Acute Lymphocytic Leukemia; Address; Antibodies; Antigen-Presenting Cells; B-Cell Acute Lymphoblastic Leukemia; Biological Assay; Biological Response Modifiers; CAR T cell therapy; Calcineurin; Cell Death; Cell Line; Cell-Mediated Cytolysis; Cells; Cellular immunotherapy; Cessation of life; Child; Childhood; Childhood Leukemia; Clinical Trials; Complement; Cyclosporine; Data; Diagnosis; Disease; Enzyme-Linked Immunosorbent Assay; Exhibits; Flow Cytometry; Genetic; Goals; Hematologic Neoplasms; Hodgkin Disease; Human; Human Cell Line; Immune; Immune Evasion; Immune response; Immune system; Immunity; Immunocompetent; Immunocompromised Host; Immunologic Surveillance; Immunotherapy; In Vitro; Inflammatory; Innate Immune System; Interferon Type II; Interleukin-12; Interleukins; Knock-out; Knockout Mice; Leukemic Cell; Licensing; Malignant Childhood Neoplasm; Mature B-Lymphocyte; Mediating; Methods; Modeling; Mus; NK Cell Activation; Natural Immunity; Natural Killer Cells; Outcome; PPP3CA gene; Patients; Peptides; Pharmacology; Play; Production; Prognosis; Proliferating; Protein Serine/Threonine Phosphatase; Rag1 Mouse; Recombinant Interleukin-12; Recombinants; Regulation; Relapse; Role; Sampling; Signal Transduction; Solid Neoplasm; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Transplantation; Western Blotting; Wild Type Mouse; acute lymphoblastic leukemia cell; adaptive immunity; cancer cell; chemotherapy; cytokine; cytotoxicity; defined contribution; human model; immune checkpoint; immunogenicity; improved; in vivo; in vivo Model; inhibitor; knock-down; leukemia; leukemogenesis; mortality; novel; novel therapeutics; nuclear factors of activated T-cells; overexpression; pathogen; response; small hairpin RNA; targeted treatment; therapy development; Acute Lymphocytic Leukemia; Address; Antibodies; Antigen-Presenting Cells; B-Cell Acute Lymphoblastic Leukemia; Biological Assay; Biological Response Modifiers; CAR T cell therapy; Calcineurin; Cell Death; Cell Line; Cell-Mediated Cytolysis; Cells; Cellular immunotherapy; Cessation of life; Child; Childhood; Childhood Leukemia; Clinical Trials; Complement; Cyclosporine; Data; Diagnosis; Disease; Enzyme-Linked Immunosorbent Assay; Exhibits; Flow Cytometry; Genetic; Goals; Hematologic Neoplasms; Hodgkin Disease; Human; Human Cell Line; Immune; Immune Evasion; Immune response; Immune system; Immunity; Immunocompetent; Immunocompromised Host; Immunologic Surveillance; Immunotherapy; In Vitro; Inflammatory; Innate Immune System; Interferon Type II; Interleukin-12; Interleukins; Knock-out; Knockout Mice; Leukemic Cell; Licensing; Malignant Childhood Neoplasm; Mature B-Lymphocyte; Mediating; Methods; Modeling; Mus; NK Cell Activation; Natural Immunity; Natural Killer Cells; Outcome; PPP3CA gene; Patients; Peptides; Pharmacology; Play; Production; Prognosis; Proliferating; Protein Serine/Threonine Phosphatase; Rag1 Mouse; Recombinant Interleukin-12; Recombinants; Regulation; Relapse; Role; Sampling; Signal Transduction; Solid Neoplasm; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Transplantation; Western Blotting; Wild Type Mouse; acute lymphoblastic leukemia cell; adaptive immunity; cancer cell; chemotherapy; cytokine; cytotoxicity; defined contribution; human model; immune checkpoint; immunogenicity; improved; in vivo; in vivo Model; inhibitor; knock-down; leukemia; leukemogenesis; mortality; novel; novel therapeutics; nuclear factors of activated T-cells; overexpression; pathogen; response; small hairpin RNA; targeted treatment; therapy development

Project Summary Leukemia is the most common cancer in children, and although prognosis has improved over the last several decades, the mortality rate is still 15-20% in children diagnosed with acute lymphoblastic leukemia (ALL). Consequently, leukemia is a leading cause of disease-related deaths in children. Although new therapies have shown the potency of the immune system for leukemia elimination, much is still unknown regarding immune escape mechanisms in leukemia. Previous studies have implicated calcineurin (Cn), a serine threonine phosphatase, in the promotion of leukemogenesis in T-ALL, yet its role in B cell-ALL (B-ALL) has not been well established. Our lab discovered extended survival in immune-competent mice engrafted with leukemia in which the essential regulatory subunit, CnB was knocked down. Yet, survival was not as prolonged in Rag1-/- mice (lacking mature B and T cells; thus, no adaptive immunity) engrafted with calcineurin-deficient leukemia (shCnB). These data implicate Cn as an important mediator of immune evasion during leukemia progression. To identify downstream effectors of Cn-dependent immune evasion, we defined the cytokine profiles from control leukemia (shNS) and shCnB cells and found that IL-12 secretion in shCnB leukemia was significantly increased compared to shNS leukemia. IL-12 is an interleukin produced by antigen-presenting cells that induces interferon-gamma (IFN-γ) production by both T and natural killer (NK) cells, resulting in cytolytic killing of cancer cells and other pathogens. To further explore this finding, WT and Rag1-/- mice were engrafted with B-ALL and administered recombinant IL-12 (rIL-12). Both WT and Rag1-/- exhibited a prolonged survival with rIL-12 administration. This points to a clear role of the innate immune system in immune surveillance of leukemia. NK cells have a major role in innate immunity thus I intend to test the hypothesis that IL-12 secretion is regulated by Cn in B-ALL cells and also potently activates NK cells resulting in leukemia clearance. In Aim 1, we will determine how Cn regulates IL-12 secretion in B- ALL cells by using pharmacologic and genetic methods to inhibit Cn to explore IL-12 secretion and also its downstream target NFAT in both human and murine B-ALL cells and also primary B-ALL patient samples. In Aim 2, we will define the contributions of NK cells to anti-leukemia immunity using models of NK deficiency and complementation assays. NK cell activation in response to shNS and shCnB leukemia cells will also be examined using cytotoxicity assays. Our goal is to better understand immune evasion in hematological malignancies to develop novel immunotherapies to control leukemia.

项目摘要 白血病是儿童中最常见的癌症，尽管预后有所改善 几十年来，被诊断出患有急性淋巴细胞白血病的儿童的死亡率仍然为15-20％（全部）。 因此，白血病是儿童与疾病有关的主要原因。虽然新疗法 已经显示了消除白血病的免疫学系统的效力，关于 白血病中的免疫逃生机制。先前的研究已经实施了钙调神经酶（CN），这是一系列的 苏氨酸磷酸酶，在T-ALL中促进白血病发生时，其在B细胞中的作用（B-All）具有 还没有建立。我们的实验室发现了与免疫能力的小鼠的延长生存期 基本调节亚基CNB的白血病被击倒。但是，生存并不那么 长时间在rag1 - / - 小鼠（缺乏成熟的B和T细胞；因此，没有适应性免疫史） 缺乏钙的白血病（SHCNB）。这些数据暗示CN是免疫的重要介体 白血病进展过程中的逃避。为了确定CN依赖性免疫进化的下游效应， 我们定义了对照白血病（SHN）和SHCNB细胞的细胞因子谱，发现IL-12分泌 与SHNS白血病相比，在SHCNB中，白血病显着增加。 IL-12是白介素产生的 通过抗原呈递细胞，影响干扰素 - γ（IFN-γ）的产生。 （NK）细胞，导致癌细胞和其他病原体的细胞溶解杀死。为了进一步探索这一发现，WT 将RAG1 - / - 小鼠与B-ALL和施用的重组IL-12（RIL-12）植入。 wt和 RAG1 - / - 通过RIL-12给药暴露了长时间的存活率。这表明先天的明确作用 白血病免疫监测中的免疫系统。 NK细胞在先天免疫中具有重要作用 打算检验以下假设：IL-12分泌受B-All细胞中CN调节，并且也可能激活 NK细胞导致白血病清除。在AIM 1中，我们将确定CN如何调节B-中的IL-12分泌 通过使用药物学和遗传学方法来抑制CN探索IL-12分泌及其的所有细胞 人和鼠类B-所有细胞的下游靶NFAT以及原发性B-ALL患者样品。 在AIM 2中，我们将使用NK缺乏模型来定义NK细胞对抗白血病免疫的贡献 和完成分析。响应SHN和SHCNB白血病细胞的NK细胞激活也将是 使用细胞毒性评估检查。我们的目标是更好地了解血液学中的免疫驱逐 开发新型免疫疗法以控制白血病的损害。