Nitric oxide immune dependent resistance mechanisms to anti-PD-1 therapy

一氧化氮免疫依赖性抗 PD-1 疗法的耐药机制

• 批准号: 10214904
• 负责人: JOSEPH MARKOWITZ
• 金额: $ 24.88万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214904
• 关键词: Antigen Presentation; Antigen-Presenting Cells; Award; Basic Science; Bioinformatics; Biological Response Modifiers; Biostatistics Core; CD4 Positive T Lymphocytes; Cancer Center; Cells; Cessation of life; Clinical; Clinical Trials; Critical Pathways; Cutaneous Melanoma; Data; Dendritic Cells; Development; Doctor of Philosophy; Ensure; Environment; Equine mule; Event; Faculty; Failure; Flow Cytometry; Foundations; Generations; Human; Immune; Immune Tolerance; Immune checkpoint inhibitor; Immune response; Immunologic Surveillance; Immunology; Immunosuppressive Agents; Immunotherapeutic agent; Immunotherapy; Infrastructure; Interferon-alpha; Interferons; Janus kinase; Lead; Ligands; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Medical Oncologist; Melanoma Cell; Mentors; Mentorship; Messenger RNA; Methods; Modeling; Mus; Myeloid-derived suppressor cells; Nitrates; Nitric Oxide; Nivolumab; Oncologist; Pathway interactions; Patients; Pattern; Peripheral Blood Mononuclear Cell; Physicians; Population; Positioning Attribute; Proteins; Proteomics; Records; Regimen; Research; Research Personnel; Resistance; STAT protein; STAT1 gene; Scientist; Seasons; Series; Signal Transduction; Specimen; Suppressor-Effector T-Lymphocytes; T-Cell Receptor; T-Lymphocyte; Therapeutic; Therapeutic Intervention; Tissues; Toxic effect; Translational Research; Tumor Immunity; Tumor-infiltrating immune cells; United States; Unresectable; Up-Regulation; Work; anti-CTLA-4 therapy; anti-PD-1; anti-PD1 therapy; career; cost; effective therapy; experience; experimental study; improved; interest; melanoma; member; nitration; nitroaspirin; novel therapeutic intervention; patient response; pembrolizumab; programmed cell death ligand 1; programmed cell death protein 1; release factor; resistance mechanism; response; success; therapeutic development; tool; tumor

Project Summary/Abstract I, Joseph Markowitz, MD PhD, am a junior medical oncologist at Moffitt Cancer Center interested in developing immune-based therapeutics for the benefit of melanoma patients. Moffitt is the ideal environment for me to develop into an independent physician scientist given the dedicated Moffitt’s Melanoma and Skin Cancers Center of Excellence, superb flow cytometry core, extensive clinical trial support infrastructure, and highly supportive bioinformatics and biostatistics core. Immune-based therapy has had mounting successes in the past few years with the introduction of checkpoint inhibitors. However, many patients do not respond to therapy, and I intend to dedicate my career into turning these failures into successes. I will have the mentorship to adapt and grow with the project. My mentorship team (Drs. Mulé, Sondak, Conejo-Garcia, Tarhini, and Berglund) was carefully selected to include translational and clinical mentors to ensure that I can mature into a successful immuno- oncologist for therapeutic development in melanoma. Melanoma cells protect themselves from immune attack by releasing factors that stimulate development of immune suppressor cells. Immune suppressor cells, such as myeloid-derived suppressor cells (MDSCs), release large amounts of nitric oxide (NO) that inactivate proteins that normally help immune cells sense and respond to cancer. Hypothesis: Anti-PD-1 resistance in melanoma results, in part, from reduction of DC antigen presentation to T cells via a NO dependent mechanism. Furthermore, NO causes nitration of multiple proteins including STAT1 and NFκB as well as MHC and T-cell receptor (TCR) molecules expressed by immune cells such as dendritic cells (DC), making them less effective for transmitting signals crucial for antigen presentation and rejection of tumors. Therefore, this proposal outlines a series of experiments that will: 1) use murine and ex-vivo models to measure antigen presentation from DC to T cells to elucidate the operative mechanism(s) by which nitration of antigen presentation proteins such as STAT1 results in immune tolerance to melanoma and resistance to anti-PD-1 therapy and to follow the levels of MDSCs, nitric oxide, and nitric oxide producing MDSCs over the course of treatment using multidimensional flow cytometry, and 2) access to well-annotated tissue specimens from melanoma patients receiving anti-PD-1 treatment (unresectable stage III/IV) to quantify the effects of NO and correlate this with changes in immune cell responses to interferon, as well as mRNA and proteomics profiles. Work completed in this proposal will potentially enable the development of new therapeutic strategies to overcome the inhibitory effects of nitric oxide and enhance patient immune responses to melanoma with anti-PD-1 therapy (and potentially other immune- based therapies). In addition, this proposal may lead to new quantitative mass spectrometry and bioinformatics methods to predict who will respond to anti-PD-1 therapy and spare patients from unnecessary toxicity if they are unlikely to respond. The work will also provide me with the necessary research and clinical foundation to become an independent investigator.

项目摘要/摘要 我，约瑟夫·马科维茨（Joseph Markowitz），医学博士，是莫菲特癌症中心（Moffitt Cancer Center）的初级医学肿瘤学家 对黑色素瘤患者益处的免疫理论。莫菲特是我的理想环境 鉴于专门的莫菲特黑色素瘤和皮肤癌中心，发展成为独立的物理科学家 卓越，出色的流式细胞仪核心，广泛的临床试验支持基础设施和高度支持 生物信息学和生物统计学核心。在过去的几年中，基于免疫的疗法取得了成功 随着检查点抑制剂的引入。但是，许多患者对治疗没有反应，我打算 将我的职业转变为将这些失败变成成功。我将拥有适应和成长的精神职位 项目。我的Mentalship Team（Mulé博士，Sondak，Conejo-Garcia，Tarhini和Berglund）小心翼翼 选择包括翻译和临床指导者，以确保我可以成熟成成功的免疫 黑色素瘤热发育的肿瘤学家。黑色素瘤细胞免受免疫攻击 通过释放刺激免疫抑制细胞发展的因素。免疫抑制细胞，例如 髓样衍生的抑制细胞（MDSC），释放大量的一氧化氮（NO），使蛋白质失活 这通常有助于免疫细胞感知并应对癌症。假设：黑色素瘤中的抗PD-1抗性 结果部分从通过无依赖机制的DC抗原呈递减少到T细胞。 此外，无引起多种蛋白质的硝化，包括STAT1和NFκB以及MHC和T-Cell 免疫细胞（例如树突状细胞（DC））表达的受体（TCR）分子，使其有效降低 传输信号对于抗原表现和排斥肿瘤至关重要。因此，该提议概述了 一系列实验将：1）使用鼠和前体模型测量从DC到 T细胞阐明了操作机制，抗原表现蛋白的硝化作用（例如） STAT1导致对黑色素瘤的免疫耐受性和对抗PD-1治疗的抗性，并遵循 MDSC，一氧化氮和一氧化氮在治疗过程中使用多维流量 细胞仪和2）从接受抗PD-1的黑色素瘤患者获得通知的组织标本 治疗（不可切除的第三阶段/IV）来量化NO的影响并将其与免疫细胞的变化相关 对干扰素以及mRNA和蛋白质组学的反应。在本提案中完成的工作将 有可能使新的治疗策略的发展能够克服一氧化氮的抑制作用 并通过抗PD-1治疗增强患者对黑色素瘤的免疫调查（以及其他免疫复杂 - 基于疗法）。此外，该建议可能会导致新的定量质谱和生物信息学 预测谁将对抗PD-1疗法做出反应的方法，并使患者免于不必要的毒性 不太可能做出回应。这项工作还将为我提供必要的研究和临床基础 成为独立研究者。