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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要白血病是儿童中最常见的癌症，尽管预后在过去几年中有所改善几十年来，被诊断患有急性淋巴细胞白血病（ALL）的儿童死亡率仍然为 15-20%。尽管有新的治疗方法，但经测试，白血病是儿童疾病相关死亡的主要原因。已显示免疫系统消除白血病的效力，但仍有许多未知之处先前的研究表明钙调神经磷酸酶（Cn）是一种丝氨酸，与白血病的免疫逃逸机制有关。苏氨酸磷酸酶在 T-ALL 中促进白血病发生，但其在 B 细胞 ALL (B-ALL) 中的作用我们的实验室发现，移植了免疫活性的小鼠的存活期延长了。然而，在白血病中，重要的调节亚基 CnB 被敲除，但存活率却不尽如人意。在 Rag1-/- 小鼠（缺乏成熟的 B 和 T 细胞；因此，没有适应性免疫）中延长这些数据表明 Cn 是一种重要的免疫介质。白血病进展过程中的逃避确定 Cn 依赖性免疫逃避的下游影响，我们定义了对照白血病 (shNS) 和 shCnB 细胞的细胞因子谱，发现 IL-12 分泌与shNS白血病相比，shCnB白血病中IL-12是一种白细胞介素产生的显着增加。由抗原呈递细胞诱导 T 细胞和自然杀伤细胞产生干扰素-γ (IFN-γ) (NK) 细胞，导致癌细胞和其他病原体的细胞溶解杀伤。为了进一步探索这一发现，WT。和 Rag1-/- 小鼠被植入 B-ALL 并给予重组 IL-12 (rIL-12)。 Rag1-/- 显示 rIL-12 给药后存活时间延长，这表明先天性的作用明显。免疫系统在白血病的免疫监视中具有重要作用，因此我认为NK细胞在先天免疫中发挥着重要作用。打算检验以下假设：IL-12 分泌受 B-ALL 细胞中的 Cn 调节，并且还有效激活 NK 细胞导致白血病清除在目标 1 中，我们将确定 Cn 如何调节 B- 中的 IL-12 分泌。利用药理学和遗传学方法抑制Cn的ALL细胞探讨IL-12的分泌及其作用下游目标人类和小鼠 B-ALL 细胞以及原代 B-ALL 患者样本中的 NFAT。在目标 2 中，我们将使用 NK 缺陷模型来定义 NK 细胞对抗白血病免疫的贡献以及针对 shNS 和 shCnB 白血病细胞的 NK 细胞激活试验。我们的目标是更好地了解血液学中的免疫逃避。开发新的免疫疗法来控制白血病。