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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的：钙/钙调蛋白依赖性蛋白激酶激酶 2 (CaMKK2) 是细胞因子的重要调节因子。代谢。尽管传统上与神经元和巨噬细胞相关，但我们的数据表明 CaMKK2 也与神经元和巨噬细胞相关。在肿瘤浸润自然杀伤 (NK) 细胞中表达，可能调节这些细胞的抗肿瘤活性。先前的出版物表明，CaMKK2 表达通过以下方式增强小鼠乳腺肿瘤的生长：将肿瘤相关巨噬细胞极化为促肿瘤 M2 表型。然而，我们的初步数据表明 CaMKK2 在 NK 细胞中发挥相反的作用：CaMKK2 信号传导似乎增强 NK 细胞的存活，增殖、迁移和抗转移免疫监视。我们假设 CaMKK2 增强 NK 细胞通过激活 AKT1、AMPK 和 mTORC1 途径发挥作用。使用 CaMKK2 基因敲除小鼠模型和常见的体外测定，我们打算坚定地描述 CaMKK2 对各种 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 AKT1、AMPK 和 mTORC1 通路在 CaMKK2 介导中的作用表型。由于 CaMKK2 在肿瘤条件下选择性地在 NK 细胞中表达，因此我们计划使用 ATAC 测序以确定负责 CaMKK2 上调的途径和肿瘤因子。最后，我们将使用 Lox-Cre 系统选择性消融小鼠 NK 细胞中的 CaMKK2 并确定这种消融的总体效果关于肿瘤转移。如果 CaMKK2 的缺失被证明是有害的，我们将改造 NK 细胞系 NK-92 过度表达 CaMKK2 并在输注至具有转移性病变的小鼠体内时测定其抗肿瘤功效。多种 CaMKK2 抑制剂和降解剂正在开发用于癌症治疗，其中一些已经已被证明在小鼠乳腺癌模型中有效。如果我们证明 CaMKK2 的缺失或抑制正如假设的那样抑制 NK 细胞功能，那么我们的研究将有助于确定目标这些新药的患者群体。患有 NK 细胞敏感性肿瘤（例如 MHC I 类阴性）的患者或接受 NK 细胞介导疗法（例如单克隆抗体疗法）的患者应避免使用 CaMKK2 抑制剂。我们的研究还可能促进解耦 CaMKK2 抑制剂对 NK 细胞和其他细胞的影响的策略。细胞类型。我们相信 CaMKK2 抑制剂的大部分治疗功效来自于它们对巨噬细胞，因此应优先考虑 CaMKK2 抑制剂的巨噬细胞特异性递送策略。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。永生化人类 NK 细胞系（例如 NK-92）作为癌症疗法正在积极研究中。如果CaMKK2 表达增强 NK 细胞的抗肿瘤活性，然后可以对 NK 细胞系进行基因改造，人工合成过度表达 CaMKK2，增强 NK 细胞输注的功效。