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: 10274980
• 负责人: WEIPING ZOU
• 金额: $ 64.74万
• 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/10274980
• 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; Immunization; Immunologics; Immunology; Immunotherapeutic agent; Immunotherapy; Impairment; Interferons; Light; Link; Lytic; Malignant Neoplasms; Measurement; Mechanics; Mediating; Mediator of activation protein; 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; 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; Tumor-infiltrating immune cells; Warburg Effect; aerobic glycolysis; amino acid metabolism; base; cancer cell; cancer genetics; cancer stem cell; cancer therapy; cell killing; cytotoxic; cytotoxic CD8 T cells; effector T cell; empowered; imaging system; immune checkpoint blockade; immune resistance; interdisciplinary approach; laser tweezer; mass spectrometric imaging; neoplasm immunotherapy; neoplastic cell; novel; perforin; resistance mechanism; stem-like cell; stemness; tool; tumor; 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 免疫治疗是产生长期保护性效应的T细胞免疫，并导致肿瘤的放射治疗 癌症患者。为此，肿瘤细胞必须对CTL介导的肿瘤杀伤具有接受性和易感性。 然而，除了CTL的质量和数量外，决定肿瘤细胞对CTL- 人们对介导的肿瘤杀伤知之甚少。进行全面的分子和功能研究是必不可少的 人类肿瘤微环境中肿瘤细胞对CTL敏感性的研究。 肿瘤生物学家一直致力于研究肿瘤代谢如何使肿瘤细胞增殖、存活和 侵袭性没有全面考虑免疫的参与。最新免疫学 研究已经开始研究肿瘤新陈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亚群。然而，肿瘤代谢如何影响肿瘤细胞对CTL的接受性- 杀戮基本上是未知的。建立了光钳显微镜(OTM)技术和原子 力显微镜(AFM)用于测量细胞弹性和硬度，并应用了气流辅助 解吸电喷雾电离质谱仪(AFADESI-MSI)，我们能够初步 证明人类和小鼠乳腺肿瘤细胞摄取特定氨基酸的结果 在减少肌球蛋白II介导的收缩激活和肿瘤细胞僵硬方面。肿瘤僵硬的丧失导致 肿瘤细胞膜对穿孔素钻孔力的抵抗和CTL的孔洞形成导致T细胞受损 细胞介导的肿瘤杀伤。从机理上讲，肿瘤细胞对某些氨基酸上瘾，包括 谷氨酰胺，通过谷氨酰胺转运蛋白SLC6A14的高表达。有趣的是，肿瘤SLC6A14的表达 受低氧控制。这些数据揭示了以前未知的特定基因之间的联系机制 肿瘤微环境中的氨基酸代谢、缺氧和T细胞免疫，从而识别 癌症谷氨酰胺转运体(S)作为潜在的新的免疫代谢检查点靶点。 基于这些令人惊讶和新奇的发现，我们假设癌细胞的硬度决定了肿瘤细胞 对CTL杀伤的感受性，以及低氧和特定氨基酸摄取之间的相互作用是一个新的 肿瘤微环境中的免疫逃逸机制。我们提出了两个具体目标和八个分目标来 测试我们的中心假设，即缺氧以肿瘤氨基酸转运体为目标，以SLC6A14为例，以 改变肿瘤细胞对CTL杀伤和肿瘤免疫及治疗的敏感性。我们的具体目标是： 目的1是验证我们的假设，即低氧代谢会削弱肿瘤对CTL杀伤的耐受性。 目的2是验证我们的假设，即低氧机械地削弱肿瘤对CTL杀伤的敏感性。