CTL-killing capacity and cancer stiffness in cancer immunity and therapy

癌症免疫和治疗中的 CTL 杀伤能力和癌症硬度

• 批准号: 10548120
• 负责人: WEIPING ZOU
• 金额: $ 63.44万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-07 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10548120
• 关键词: Aerobic; Affect; Air Movements; Amino Acid Transporter; Amino Acids; Attention; Biological; Biomechanics; Breast Cancer Cell; CD8-Positive T-Lymphocytes; Cell Proliferation; Cell membrane; Cells; Characteristics; Chemoresistance; Cytoplasmic Granules; DNA Sequence Alteration; Data; Development; Effector Cell; Elasticity; Epigenetic Process; Event; FOXP3 gene; Genetic; Glutamine; Goals; Granzyme; Human; Hypoxia; Immune; Immune Evasion; Immune response; Immune system; Immunity; Immunologic Sensitization; Immunologics; Immunology; Immunotherapeutic agent; Immunotherapy; Impairment; Interferons; Link; Lytic; Malignant Neoplasms; Measurement; Mechanics; Mediating; Mediator; Membrane; Metabolic; Metabolism; Microscope; Molecular; Mus; Mutation; Myeloid-derived suppressor cells; Myosin Type II; Nature; Neoplasm Metastasis; Oncology; Pathology; Pathway interactions; Patient-Focused Outcomes; Patients; Phenotype; Predisposition; Process; Regulation; Regulatory T-Lymphocyte; Research; Resistance; Role; Scanning Probe Microscopes; Signal Pathway; Solid Neoplasm; Spectrometry, Mass, Electrospray Ionization; T-Lymphocyte; Techniques; Testing; Tissues; Tumor Immunity; Warburg Effect; aerobic glycolysis; amino acid metabolism; cancer cell; cancer genetics; cancer stem cell; cancer therapy; cell killing; cytotoxic; cytotoxic CD8 T cells; effector T cell; empowerment; imaging system; immune checkpoint blockade; immune resistance; interdisciplinary approach; laser tweezer; mass spectrometric imaging; migration; neoplasm immunotherapy; neoplastic cell; novel; optic tweezer; perforin; resistance mechanism; stem-like cell; stemness; tool; trafficking; tumor; tumor initiation; tumor metabolism; tumor microenvironment; uptake

Project Summary/Abstract: CD8+ T cells (CTLs) mediate protective tumor immunity. The goal of tumor immune therapy is to engender long-term protective effector T cell immunity and cause tumor eradiation in patients with cancer. To this end, tumor cells must be receptive and susceptible to CTL-mediated tumor killing. However, in addition to the quality and quantity of CTLs, what determines tumor cell receptivity to CTL- mediated tumor killing is poorly understood. It is essential to conduct comprehensive molecular and functional research on the nature of tumor cell receptivity to CTLs in the human tumor microenvironment. Tumor biologists have been working on how tumor metabolism enables tumor cell proliferation, survival, and invasiveness without a comprehensive consideration of the immune involvement. Recent immunological studies have started to examine how tumor metabolism affects the phenotype and function of different immune cell subsets in the tumor microenvironment. Yet, how tumor metabolism affects tumor cell receptivity to CTL- killing is basically unknown. Having established an Optical Tweezers Microscope (OTM) technique and Atomic Force Microscope (AFM) for cell elasticity and stiffness measurements and applied an airflow-assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI), we are able to preliminarily demonstrate that human and mouse breast tumor cells empowered with particular amino acid uptake resulted in reduced myosin II-mediated contractile activation and tumor cell stiffness. Loss of tumor stiffness led to tumor cell membrane resistance to perforin drilling force and pore formation by CTLs, resulting in impaired T cell-mediated tumor killing. Mechanistically, tumor cells were addicted to certain amino acids, including glutamine, via high expression of SLC6A14, a glutamine transporter. Interestingly, tumor SLC6A14 expression was controlled by hypoxia. These data reveal previously unknown mechanisms of connection between specific amino acid metabolism, hypoxia, and T cell immunity in the tumor microenvironment, thereby identifying cancer glutamine transporter(s) as a potential novel immune metabolic checkpoint target. Based on these surprising and novel findings, we hypothesize that cancer cell stiffness determines tumor cell receptivity to CTL-killing, and the interplay between hypoxia and particular amino acid uptake is a novel immune evasion mechanism in the tumor microenvironment. We propose two specific aims and 8 subaims to test our central hypothesis that hypoxia targets tumor amino acid transporters, exemplified by SLC6A14, to alter tumor cell receptivity to CTL-killing and tumor immunity and therapy. Our specific aims are: Aim 1 is to test our hypothesis that hypoxia metabolically weakens tumor receptivity to CTL-killing. Aim 2 is to test our hypothesis that hypoxia mechanically weakens tumor receptivity to CTL-killing.

项目摘要/摘要：CD8+ T细胞（CTL）介导保护性肿瘤免疫。肿瘤的目标 免疫治疗是为了产生长期保护效应T细胞免疫，并导致肿瘤消除 癌症患者。为此，肿瘤细胞必须接受并容易受到CTL介导的肿瘤杀伤的影响。 但是，除了CTL的质量和数量之外，什么决定了肿瘤细胞对CTL-的接受度 介导的肿瘤杀死知之甚少。进行全面的分子和功能至关重要 研究人类肿瘤微环境中肿瘤细胞接受性的性质。 肿瘤生物学家一直在研究肿瘤代谢如何使肿瘤细胞增殖，生存和 侵入性无需全面考虑免疫参与。最近的免疫学 研究开始研究肿瘤代谢如何影响不同免疫的表型和功能 肿瘤微环境中的细胞子集。然而，肿瘤代谢如何影响肿瘤细胞对CTL- 杀人基本上是未知的。建立了光学镊子显微镜（OTM）技术和原子 力显微镜（AFM）用于细胞弹性和刚度测量，并应用了气流辅助 解吸电喷雾电离质谱成像（Afadesi-MSI），我们能够初步 证明由特定氨基酸摄取的人类和小鼠乳腺肿瘤细胞导致 在减少肌球蛋白II介导的收缩激活和肿瘤细胞刚度中。失去肿瘤僵硬导致 肿瘤细胞膜耐药性对穿孔蛋白钻孔作用和CTL形成孔，导致T受损 细胞介导的肿瘤杀死。从机械上讲，肿瘤细胞沉迷于某些氨基酸，包括 谷氨酰胺通过SLC6A14（谷氨酰胺转运蛋白）的高表达。有趣的是，肿瘤SLC6A14表达 由缺氧控制。这些数据揭示了以前未知的特定连接机制 肿瘤微环境中的氨基酸代谢，缺氧和T细胞免疫，从而鉴定 癌症谷氨酰胺转运蛋白作为潜在的新型免疫代谢检查点目标。 基于这些令人惊讶和新颖的发现，我们假设癌细胞的刚度决定了肿瘤细胞 对CTL杀伤的接受性以及缺氧与特定氨基酸摄取之间的相互作用是一种新颖的 肿瘤微环境中的免疫逃避机制。我们提出了两个具体目标和8个subiaims 测试我们的中心假设，即缺氧靶向肿瘤氨基酸转运蛋白，以SLC6A14为例 改变肿瘤细胞的接受性对CTL杀伤和肿瘤免疫和治疗。我们的具体目的是： 目的1是测试我们的假设，即缺氧代谢削弱了肿瘤对CTL杀伤的接受能力。 目的2是测试我们的假设，即缺氧会机械地削弱肿瘤对CTL杀伤的接受能力。