The role of CaMKK2 in tumor-infiltrating natural killer cells

CaMKK2 在肿瘤浸润自然杀伤细胞中的作用

• 批准号: 10375395
• 负责人: Patrick Juras
• 金额: $ 3.88万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10375395
• 关键词: AKT1 gene; Ablation; Affect; Apoptosis; Attention; Binding; Biological Assay; Breast Cancer Model; Breast Epithelial Cells; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Cell Line; Cell Survival; Cell Therapy; Cell physiology; Cells; Cellular biology; Clinical; Cre lox recombination system; Data; Development; Disease; Engineering; Enzymes; Genetic; Genetic Engineering; Genetic Transcription; Hepatocyte; Human; Immune Evasion; Immunocompromised Host; Immunologic Surveillance; Infiltration; Investigation; Knockout Mice; Lead; Major Histocompatibility Complex; Malignant Neoplasms; Mammary Neoplasms; Mediating; Metabolic; Metabolism; Methods; Modification; Monoclonal Antibody Therapy; Mus; Natural Killer Cells; Neoplasm Metastasis; Neurons; Outcome; Pathway interactions; Patients; Pharmacology; Phenotype; Phosphotransferases; Pilot Projects; Play; Process; Proteins; Publications; Regulation; Reporting; Research; Role; Signal Pathway; Signal Transduction; Solid Neoplasm; Stress; Transfusion; Treatment Efficacy; Tumor Burden; Tumor-associated macrophages; Up-Regulation; XCL1 gene; antibody-dependent cell cytotoxicity; cancer therapy; cell motility; cell type; clinical application; conditioning; cytotoxic; cytotoxicity; engineered NK cell; fighting; genetic regulatory protein; in vitro Assay; inhibitor; insight; interest; macrophage; migration; mouse model; neoplastic cell; novel strategies; novel therapeutics; overexpression; patient population; phosphoproteomics; response; selective expression; small molecule inhibitor; therapeutic target; transcription factor; tumor; tumor growth

Abstract: Calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is an important regulator of cellular metabolism. Though classically associated with neurons and macrophages, our data show that CaMKK2 is also expressed in tumor-infiltrating natural killer (NK) cells and may regulate the anti-tumor activity of these cells. Previous publications have shown that CaMKK2 expression enhances breast tumor growth in mice by polarizing tumor-associated macrophages toward a pro-tumor M2 phenotype. However, our preliminary data suggest that CaMKK2 plays the opposite role in NK cells: CaMKK2 signaling seems to enhance NK cell survival, proliferation, migration, and anti-metastatic immune surveillance. We postulate that CaMKK2 enhances NK cell functions by activating the AKT1, AMPK, and mTORC1 pathways. Using a genetic knock-out mouse model of CaMKK2 and common in vitro assays, we intend to firmly delineate the effects of CaMKK2 on a variety of NK cellular functions. Using small-molecule inhibitors or activators, we will determine the role of the AKT1, AMPK, and mTORC1 pathways in CaMKK2-mediated phenotypes. Because CaMKK2 is selectively expressed in NK cells under tumor conditions, we plan to use ATAC sequencing to identify pathways and tumor factors responsible for CaMKK2 upregulation. Finally, we will use the Lox-Cre system to selectively ablate CaMKK2 in murine NK cells and determine the overall effect of this ablation on tumor metastasis. If the loss of CaMKK2 proves detrimental, we will engineer the NK cell line NK-92 to overexpress CaMKK2 and assay its anti-tumor efficacy when transfused into mice with metastatic lesions. A variety of CaMKK2 inhibitors and degraders are being developed for cancer treatment, and some have already proven efficacious in murine breast cancer models. If we demonstrate that loss or inhibition of CaMKK2 suppresses NK cell function as hypothesized, then our research will be instrumental in determining the target patient population for these new drugs. Patients with NK-cell sensitive tumors (e.g. MHC Class I-negative) or patients taking NK-cell mediated therapies (e.g. monoclonal antibody therapies) should avoid CaMKK2 inhibitors. Our research may also prompt strategies for decoupling the effects of CaMKK2 inhibitors on NK cells and other cell types. We believe that much of the therapeutic efficacy of CaMKK2 inhibitors comes from their effects on macrophages, so macrophage-specific delivery strategies for CaMKK2 inhibitors should be prioritized. Furthermore, our research suggests a novel strategy for enhancing the activity of NK cell transfusions. Immortalized human NK cell lines such as NK-92 are under active investigation as cancer therapies. If CaMKK2 expression enhances NK cell anti-tumor activity, then an NK cell line could be genetically modified to artificially overexpress CaMKK2, enhancing the efficacy of the NK cell transfusion.

摘要： 钙/钙调蛋白依赖性蛋白激酶激酶2（CaMKK 2）是一种重要的细胞凋亡调节因子， 新陈代谢.虽然CaMKK 2与神经元和巨噬细胞有着传统的联系，但我们的数据显示，CaMKK 2也是一种神经元和巨噬细胞。 在肿瘤浸润性自然杀伤（NK）细胞中表达，并可调节这些细胞的抗肿瘤活性。 先前的出版物已经表明，CaMKK 2表达通过以下方式促进小鼠乳腺肿瘤生长： 使肿瘤相关巨噬细胞向促肿瘤M2表型极化。然而，我们的初步数据显示， 提示CaMKK 2在NK细胞中起相反的作用：CaMKK 2信号传导似乎增强NK细胞存活， 增殖、迁移和抗转移免疫监视。我们推测CaMKK 2可以增强NK细胞的杀伤活性， 通过激活AKT 1、AMPK和mTORC 1通路发挥作用。 使用CaMKK 2基因敲除小鼠模型和常见的体外试验，我们打算坚定地 描述了CaMKK 2对多种NK细胞功能的影响。使用小分子抑制剂或 激活剂，我们将确定AKT 1，AMPK和mTORC 1通路在CaMKK 2介导的细胞凋亡中的作用。 表型由于CaMKK 2在肿瘤条件下选择性地在NK细胞中表达，我们计划使用ATAC 测序以鉴定负责CaMKK 2上调的途径和肿瘤因子。最后，我们将使用 Lox-Cre系统选择性消融鼠NK细胞中的CaMKK 2并确定该消融的总体效果 对肿瘤转移的影响如果CaMKK 2的丢失被证明是有害的，我们将改造NK细胞系NK-92， 过量表达CaMKK 2并测定其在输注到具有转移性病变的小鼠中时的抗肿瘤功效。 正在开发各种CaMKK 2抑制剂和降解剂用于癌症治疗，其中一些已经 已经在小鼠乳腺癌模型中证明有效。如果我们证明CaMKK 2的缺失或抑制 抑制NK细胞功能的假设，那么我们的研究将有助于确定目标 这些新药的患者群体。NK细胞敏感性肿瘤患者（例如，MHC I类阴性）或 服用NK细胞介导疗法（例如单克隆抗体疗法）的患者应避免CaMKK 2抑制剂。 我们的研究还可能提示将CaMKK 2抑制剂对NK细胞和其他细胞的影响解耦的策略。 细胞类型。我们认为，CaMKK 2抑制剂的大部分治疗功效来自于它们对 巨噬细胞，因此CaMKK 2抑制剂的巨噬细胞特异性递送策略应优先考虑。 此外，我们的研究提出了一种增强NK细胞输注活性的新策略。 永生化人NK细胞系如NK-92作为癌症疗法正在积极研究中。如果CaMKK 2 表达增强NK细胞的抗肿瘤活性，那么NK细胞系可以被遗传修饰以人工地 过表达CaMKK 2，增强NK细胞输注的功效。