Mitochondrial Regulation of Interferon Response in Melanoma

黑色素瘤中干扰素反应的线粒体调节

• 批准号: 10752523
• 负责人: Melissa Johnson
• 金额: $ 4.05万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752523
• 关键词: ATAC-seq; Acute; Adaptive Immune System; Affect; Antigen Presentation Pathway; Antioxidants; Antitumor Response; Autoimmune; CD8-Positive T-Lymphocytes; CRISPR screen; Cellular Metabolic Process; ChIP-seq; Chromatin; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Dependence; Development; Down-Regulation; Effectiveness; Electron Transport; Event; Expression Profiling; Fatty Acids; Fluorescence Microscopy; Heart; Immune Evasion; Immune response; Immune system; Immunosuppression; Immunotherapy; In Vitro; Innate Immune System; Interferon Type I; Interferon Type II; Interferon alpha; Interferon-beta; Interferons; Knock-out; Link; Major Histocompatibility Complex; Malignant Neoplasms; Mediating; Melanoma Cell; Membrane Potentials; Metabolic; Metabolism; Mitochondria; Mus; Oxidative Phosphorylation; Pathway interactions; Phenotype; Phosphorylation; Play; Process; Production; Proteins; RNA; Reaction; Reactive Oxygen Species; Regulation; Role; Screening Result; Signal Pathway; Signal Transduction; Skin Cancer; Surface; Testing; Therapeutic; Transcriptional Activation; Tumor Immunity; Up-Regulation; Western Blotting; Work; anti-tumor immune response; cancer cell; cancer immunotherapy; cancer therapy; cell type; desensitization; fatty acid metabolism; fatty acid oxidation; improved; in vivo; insight; melanoma; metabolomics; mitochondrial membrane; mitochondrial metabolism; neoplastic cell; novel; pharmacologic; programmed cell death ligand 1; response; therapy development; transcription factor; transcriptomics; tumor; tumor metabolism; tumor microenvironment; tumor progression; tumor-immune system interactions

PROJECT SUMMARY Interferons (IFNs) are central orchestrators of tumor immunity and can elicit both pro-tumor and anti-tumor responses depending on the cancer cell type, the type of IFN produced (e.g. a, b or g), duration of the signal, and other factors in the tumor microenvironment (TME). In their anti-tumor role, IFNs induce expression of major histocompatibility complex I (MHC-I) on the surface of tumor cells that mediates CD8+ T cell recognition and killing. Paradoxically, IFNs also upregulate expression of CD8+ T cell inhibitory surface molecules such as programmed death ligand 1 (PD-L1) to temper the immune response and avoid an autoimmune reaction. Thus, IFNs play a dual and opposing role in cancer development, making a more complete understanding of the contexts in which their pro- or anti-tumor functions predominate important for effective cancer therapy development. Immune evasion can occur when malignant cells lose MHC-I and antigen processing and presentation (APP) machinery or otherwise become desensitized to IFN signaling. Thus, finding ways to reinvigorate these pathways has significant therapeutic potential. Interestingly, recent work from the sponsor’s lab and others has shown that mitochondrial electron transport chain activity is required for IFN-induced MHC-I expression. In addition, preliminary data show that chronic IFN stimulation in vitro reduces mitochondrial oxidative phosphorylation (OXPHOS) in melanoma cells, suggesting that IFN signaling can also influence tumor mitochondrial metabolism. To probe this novel regulatory link between the metabolic state of tumor cells and their responses to IFN, a metabolism-targeted CRISPR knock-out screen was performed in mouse melanoma cells. Results from this screen not only confirmed a requirement for mitochondrial OXPHOS in regulating IFN signaling, but also implicated fatty acid metabolism and ROS. Based on these preliminary data, it is proposed that mitochondrial OXPHOS, specifically mitochondrial fuel utilization and ROS production, can directly regulate key IFN signaling steps, and that chronic IFN exposure leads to changes in mitochondrial metabolism that facilitate immune evasion in melanoma. This overall hypothesis will be tested through completion of two specific aims. Aim 1 is to determine which steps in the IFN signaling pathway are subject to mitochondrial OXPHOS- mediated regulation and the precise mitochondrial metabolic signals involved. Aim 2 is to determine how reduced mitochondrial OXPHOS and/or increased mitochondrial ROS are generating a suppressive tumor microenvironment and if chronic type I IFN signaling elicits similar immunosuppressive effects as chronic type II IFN exposure. This project will provide important new insights into the relationship between mitochondrial metabolism and IFN responses during tumor progression that might be exploited to improve or reactivate anti- tumor immune responses for better cancer treatment and augment cancer immunotherapy.

项目总结 干扰素(IFN)是肿瘤免疫的中枢协调器，既能诱导促肿瘤免疫，又能诱导抗肿瘤免疫 根据癌细胞类型、产生的干扰素的类型(例如A、B或G)、信号的持续时间、 以及肿瘤微环境(TME)中的其他因素。在其抗肿瘤作用中，IFN诱导主要 肿瘤细胞表面的组织相容性复合体I(MHC-I)介导CD8+T细胞识别和 杀戮。矛盾的是，IFN还上调CD8+T细胞抑制表面分子的表达，如 程序化死亡配体1(PD-L1)，以缓和免疫反应，避免自身免疫反应。因此， IFN在癌症的发展中扮演着双重和相反的角色，使人们对 它们的促肿瘤或抗肿瘤功能对有效的癌症治疗非常重要的背景 发展。当恶性细胞失去MHC-I和抗原处理时，就会发生免疫逃避 呈现(APP)机制或以其他方式对干扰素信号脱敏。因此，想方设法 重振这些通路具有显著的治疗潜力。有趣的是，赞助商的最新作品 Lab等人已经证明，干扰素诱导的MHC-I需要线粒体电子传输链的活性 表情。此外，初步数据显示，体外慢性干扰素刺激会减少线粒体 黑色素瘤细胞中的氧化磷酸化(OXPHOS)，提示干扰素信号也可以影响肿瘤 线粒体代谢。为了探索肿瘤细胞代谢状态和肿瘤细胞之间的这种新的调控联系 他们对干扰素的反应是在小鼠黑色素瘤中进行的，这是一种针对代谢的CRISPR基因敲除筛查 细胞。这一筛选的结果不仅证实了线粒体OXPHOS在调节干扰素方面的需求 信号转导，但也与脂肪酸代谢和ROS有关。基于这些初步数据，提出了 线粒体OXPHOS，特别是线粒体的燃料利用和ROS的产生，可以直接调节 关键的干扰素信号步骤，以及长期接触干扰素会导致线粒体代谢的变化，从而 促进黑色素瘤的免疫逃逸。这一总体假设将通过完成两个具体的 目标。目的1是确定干扰素信号通路中的哪些步骤受到线粒体OXPHOS的影响。 介导的调节和精确的线粒体代谢信号。目标2是确定如何减少 线粒体OXPHOS和/或线粒体ROS增加正在产生抑制性肿瘤 微环境和慢性I型干扰素信号诱导的免疫抑制效应与慢性II型相似 干扰素暴露。这一项目将为线粒体之间的关系提供重要的新见解。 肿瘤进展过程中的代谢和干扰素反应可能被利用来改善或重新激活抗- 肿瘤免疫反应，以更好地治疗癌症，并加强癌症免疫治疗。