Metabolic adaptation enables cisplatin resistance and inhibits tumor immunity

代谢适应使顺铂耐药并抑制肿瘤免疫

• 批准号: 10707169
• 负责人: VLAD C SANDULACHE
• 金额: $ 25.6万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707169
• 关键词: Ablation; Aggressive behavior; Amino Acids; Anabolism; Automobile Driving; Biological Markers; Carbon; Catabolism; Cell Line; Cell Survival; Cells; Chemicals; Cisplatin; Clinic; Clinical; Coupled; Data; Development; Effectiveness; Enzymes; Equilibrium; FADH2; GPX2 gene; Generations; Genetic Transcription; Genomics; Genotoxic Stress; Glucose; Glutamine; Glutathione; Head and Neck Squamous Cell Carcinoma; Human; Image; Immune; Immune checkpoint inhibitor; Immunocompetent; Immunologic Surveillance; Immunologics; In Vitro; Incidence; Individual; Link; Lung; Malignant Neoplasms; Measurement; Measures; Mediating; Metabolic; Metabolism; Mitochondria; Modeling; Molecular Target; Myeloid-derived suppressor cells; NF-kappa B; Natural regeneration; Paracrine Communication; Pathway interactions; Patients; Peroxidases; Peroxides; Phenotype; Platinum; Platinum Compounds; Population; Pre-Clinical Model; Production; Prostaglandin Production; Prostaglandins; Research; Resistance; Resistance development; Role; Signal Transduction; Solid Neoplasm; Stress; Testing; Time; Treatment Failure; Tumor Immunity; Tumor-Infiltrating Lymphocytes; Work; amino acid metabolism; cancer type; chemokine; chemotherapy; cytokine; cytotoxic; defined contribution; design; disorder control; experimental study; immunoregulation; in vivo; knock-down; mouse model; neoplastic cell; overexpression; patient biomarkers; patient response; permissiveness; personalized approach; rapid detection; response; small hairpin RNA; therapy resistant; treatment response; tumor; tumor behavior; tumor growth; tumor-immune system interactions

Project 1 SUMMARY A clearer understanding of how tumor cells survive the stress of platinum agents and evolve into therapy resistant populations is essential to overcome treatment failure and maximize both disease control and survival. Our data demonstrate that HNSCC cell lines with acquired cisplatin resistance reduce glycolytic and mitochondrial energy production while increasing carbon flux into anabolic pathways. This results in an enhanced reductive potential (glutathione, NAD(P)H, FADH2) via both glucose and glutamine catabolism coupled to increased glutathione (GS) peroxidase 2 (GPX2) activity coordinated at a genomic and transcriptional level, partially through the KEAP1-NRF2 pathway. Hyperactivation of GPX2 concomitantly likely inhibits NF-κB activation, decreasing chemokine and prostaglandin production by tumor cells— leading to a suppressive tumor immune microenvironment (TIME) enriched for myeloid derived suppressor cells (MDSCs) and depleted of cytotoxic tumor infiltrating lymphocytes (TILs). It is our central hypothesis that this metabolic adaptation is a permissive and required step for acquisition of cisplatin resistance. We further postulate that this metabolic shift transitions some tumors to an immunologically silent phenotype, which reduces immune surveillance to compound the aggressive behavior and cross-therapy resistance of CDDP-treated tumors We will first define the critical metabolic steps required for generation of an enhanced reductive state that supports cisplatin resistance in Aim 1. We will utilize in vitro and in vivo orthotopic HNSCC models to measure the contribution of glucose and glutamine to GS synthesis in cisplatin resistant tumors and, using chemical inhibition coupled to shRNA blockade of individual transporters and enzymes, identify the critical rate limiting metabolic steps for GS synthesis and cisplatin resistance in HNSCC. In Aim 2 we will determine how GS synthesis and utilization (by GPX2 and non GPX means) are coordinated transcriptionally at least in part through Nrf2 in order to support cisplatin resistance. In Aim 3 we will test the impact of GS metabolism via canonical (e.g. NF-kB) and metabolic paracrine signaling on development of a suppressive TIME. Completion of the proposed experiments will: 1) identify suitable targets for ablation of the enhanced reductive state driving cisplatin resistance in HNSCC and 2) identify metabolic biomarkers which can be coupled to 13C flux imaging-based measurements to generate real-time readout of tumor treatment response, and inform a more personalized approach to targeting metabolism to overcome CDDP resistance. By identifying the critical mechanistic underpinnings of metabolic adaptation, we can generate a paradigm shift in our capability to both rapidly detect acquisition of resistance to genotoxic stress and to overcome it using multiple clinically viable approaches. It will further shed light on how acquisition of cisplatin resistance can impact response to immunomodulatory approaches such as immune checkpoint inhibitors (ICIs) currently being combined with chemotherapy in the clinic.

项目1概要 更清楚地了解肿瘤细胞如何在铂类药物的压力下生存并进化成 治疗耐药人群对于克服治疗失败和最大限度地控制疾病和 生存。我们的数据表明，具有获得性顺铂耐药性的 HNSCC 细胞系会减少糖酵解和 线粒体能量产生，同时增加进入合成代谢途径的碳通量。这导致 通过葡萄糖和谷氨酰胺分解代谢增强还原电位（谷胱甘肽、NAD(P)H、FADH2） 与基因组协调的谷胱甘肽 (GS) 过氧化物酶 2 (GPX2) 活性增加相结合 转录水平，部分通过KEAP1-NRF2途径。 GPX2 同时过度激活 可能抑制 NF-κB 激活，减少肿瘤细胞产生的趋化因子和前列腺素——领先 富含骨髓源性抑制细胞的抑制性肿瘤免疫微环境 (TIME) (MDSC) 并耗尽细胞毒性肿瘤浸润淋巴细胞 (TIL)。我们的中心假设是，这 代谢适应是获得顺铂耐药性的允许且必需的步骤。我们 进一步假设这种代谢转变将一些肿瘤转变为免疫沉默表型， 这减少了免疫监视，从而加剧了攻击行为和交叉治疗耐药性 CDDP治疗的肿瘤 我们将首先定义生成增强还原态所需的关键代谢步骤 支持目标 1 中的顺铂耐药性。我们将利用体外和体内原位 HNSCC 模型来 测量葡萄糖和谷氨酰胺对顺铂耐药肿瘤中 GS 合成的贡献，并使用 化学抑制与单个转运蛋白和酶的 shRNA 阻断相结合，确定了关键的 HNSCC 中 GS 合成和顺铂耐药性的限速代谢步骤。在目标 2 中，我们将 确定 GS 合成和利用（通过 GPX2 和非 GPX 方式）如何在转录上协调 至少部分通过 Nrf2 来支持顺铂耐药。在目标 3 中，我们将测试 GS 的影响 通过规范（例如 NF-kB）的代谢和代谢旁分泌信号对抑制性药物的发展 时间。 完成所提出的实验将：1）确定合适的消融目标 增强的还原态驱动 HNSCC 中的顺铂耐药性，2) 识别代谢生物标志物，这些标志物可以 与基于 13C 通量成像的测量相结合，生成肿瘤治疗的实时读数 响应，并提供更个性化的方法来靶向代谢以克服 CDDP 耐药性。 通过确定代谢适应的关键机制基础，我们可以生成一个范例 我们快速检测对基因毒性应激的抵抗力的获得并克服它的能力的转变 使用多种临床可行的方法。这将进一步阐明如何收购顺铂 耐药性可能会影响对免疫调节方法（例如免疫检查点抑制剂）的反应 （ICIs）目前在临床上与化疗相结合。