Dissecting innate immune mechanisms of resistance to checkpoint blockade therapy in bladder cancer

剖析膀胱癌检查点阻断治疗耐药的先天免疫机制

• 批准号: 10669596
• 负责人: Michelle Alyssa Tran
• 金额: $ 4.61万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-11 至 2026-07-10
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10669596
• 关键词: Accounting; Antigen Presentation; Atlases; Autologous; Biological; Bladder Neoplasm; Blood; CD8-Positive T-Lymphocytes; Cancer Patient; Cell physiology; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Cessation of life; Clinical; Clinical Trials; Coculture Techniques; Coin; Colon Carcinoma; Data; Exclusion; Genes; Genetic Transcription; Genomics; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Hypoxia; Immune; Immunotherapy; In Vitro; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-1 beta; Link; Macrophage; Macrophage Colony-Stimulating Factor; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mediating; Mediator; Modeling; Molecular; Myelogenous; Myeloid Cells; Outcome; PD-1/PD-L1; Pathogenesis; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Play; Population; Primary Neoplasm; Production; Proteins; Proteomics; Publishing; Research; Resistance; Resolution; Resources; Role; Shapes; Signal Transduction; Specimen; System; T-Lymphocyte; Testing; Tumor Immunity; Tumor Promotion; United States; Urine; Work; adaptive immune response; adaptive immunity; angiogenesis; anti-PD1 antibodies; chemokine; clinical biomarkers; clinical predictors; cohort; combinatorial; complement pathway; cytokine; drug candidate; drug testing; exhaustion; experimental study; follow-up; genetic signature; immune checkpoint blockade; immunoregulation; improved; inflammatory modulation; inhibitor; innate immune mechanisms; innate immune pathways; insight; malignant breast neoplasm; marenostrin; monocyte; novel strategies; patient subsets; peripheral blood; predictive marker; predictive signature; resistance gene; resistance mechanism; response; restraint; single cell technology; single-cell RNA sequencing; tool; transcriptome sequencing; transcriptomics; treatment strategy; tumor; tumor growth; tumor progression

PROJECT SUMMARY Bladder cancer is the fifth most common cancer in the United States, accounting for around 47 deaths per day. Promisingly, five PD-1/PD-L1 immune checkpoint blockade (ICB) therapies were approved for bladder cancer in 2016. Although these ICB treatments have achieved durable clinical responses in a subset of patients (15-25%), the majority of patients have still not benefitted from this therapy. This clinical urgency to extend the benefits of ICB to more patients has led to a need to investigate tumor intrinsic mechanisms underlying resistance. Tumor- promoting inflammation, a hallmark of cancer pathogenesis, is known to contribute to cancer growth in multiple ways including restraining antitumor immunity. We discovered a gene signature from pre-treatment tumor associating with myeloid cells that is enriched in inflammation and innate immune genes and predictive of poor ICB outcomes and survival in two ICB clinical trials. I plan to follow up on this work and dissect the innate immune landscape of bladder cancer and investigate mechanisms of myeloid-cell mediated resistance to ICB therapy. Aim 1 seeks to define the landscape of untreated bladder tumors and provide insight into the immune cell subsets underlying ICB resistance. I will construct a transcriptomic and molecular atlas of bladder cancer at a single-cell resolution, a resource that does not currently exist. I will build atlases of patients’ tumor, blood, and urine using single-cell RNA sequencing, Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITEseq), spatial transcriptomics, and O-link proteomics and analyze them using Seurat and other R-based tools. I plan to resolve myeloid cells expressing this resistant gene signature and define their cellular interactions. In Aim 2, I will delve into the transcriptional pathways in myeloid cells that are contributing to ICB resistance. We have identified NLRP3 inflammasome activation and IL-1β signaling in tumor monocyte-macrophages (mono- MΦs) as candidate pathways promoting tumor inflammation and progression. I will model these mono-MΦs by differentiating peripheral blood monocytes into MΦ using GM-CSF and M-CSF under hypoxic conditions with IL- 1β and NLRP3 inflammasome activators., I will test effects on adaptive immunity by co-culturing these mono- MΦs with activated autologous CD8+ T cells. I will also use this model to test drug candidates known to modulate IL-1β and NLRP3 inflammasome activity as potential combinatorial treatments with ICB in bladder cancer. This proposal combines direct ex vivo single cell genomics with in vitro functional experiments for a thorough interrogation of the innate immune contribution to ICB resistance in bladder cancer. Combined, these aims will elucidate innate immune pathway driven resistance to PD-1/PD-L1 ICB therapy in bladder cancer, which can be used to identify critical predictive clinical biomarkers and inform new combinatorial treatment strategies.

项目摘要 膀胱癌是美国第五大最常见的癌症，每天约为47例死亡。 有希望的是，批准了五个PD-1/PD-L1免疫障碍物（ICB）疗法用于Bladeder Cancer 2016年。尽管这些ICB治疗已在一部分患者中达到了持久的临床反应（15-25％），但 大多数患者仍然没有从这种疗法中受益。这种临床紧迫性，以扩大 ICB对更多的患者导致需要研究抗抗性的肿瘤固有机制。瘤- 促进炎症是癌症发病机理的标志，已知有助于多种 包括限制抗肿瘤免疫的方式。我们从治疗肿瘤中发现了一个基因签名 与富含炎症和先天免疫原的髓样细胞相关联，可预测不良 ICB的结果和生存在两项ICB临床试验中。我计划跟进这项工作并剖析先天免疫 膀胱癌的景观并研究了髓样细胞介导的对ICB治疗的抗性的机制。 AIM 1旨在定义未处理的膀胱肿瘤的景观，并提供有关免疫球的洞察力 ICB电阻的基础集。我将在A的膀胱癌的转录组和分子图中构建 单细胞分辨率，当前不存在的资源。我将建立患者肿瘤，血液和 使用单细胞RNA测序，转录组和表位的细胞索引尿液通过测序 （CITESEQ），空间转录组学和O-Link蛋白质组学，并使用Seurat和其他基于R 工具。我计划解决表达这种抗性基因特征并定义其细胞相互作用的髓样细胞。 在AIM 2中，我将深入研究有助于ICB抗性的髓样细胞的转录途径。我们 已经确定了NLRP3炎性体激活和IL-1β信号传导 MφS）作为促进肿瘤注射和进展的候选途径。我将通过 在低氧条件下，使用GM-CSF和M-CSF将外周血单核细胞区分为Mφ 1β和NLRP3炎性体激活剂。 具有活化自体CD8+ T细胞的Mφ。我还将使用此模型测试已知可以调节的候选药物 IL-1β和NLRP3炎性体活性作为膀胱癌中ICB的潜在组合治疗。 该建议将直接的离体单细胞基因组与体外功能实验结合在一起，以进行彻底 对膀胱癌对ICB抗性的先天免疫贡献的询问。结合在一起，这些目标将 阐明先天免疫途径驱动对膀胱癌中PD-1/PD-L1 ICB治疗的抗性，这可以是 用于确定关键的预测性临床生物标志物并为新的组合治疗策略提供信息。