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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要膀胱癌是美国第五大常见癌症，每天约有 47 人死亡。令人鼓舞的是，五种 PD-1/PD-L1 免疫检查点阻断 (ICB) 疗法在 2019 年被批准用于治疗膀胱癌 2016. 尽管这些 ICB 治疗在一小部分患者 (15-25%) 中取得了持久的临床反应，大多数患者仍未从这种疗法中受益，临床迫切需要扩大这种疗法的益处。对更多患者进行 ICB 导致需要研究肿瘤耐药性的内在机制。促进炎症是癌症发病机制的一个标志，已知会导致多种癌症的生长我们发现了治疗前肿瘤的基因特征。与富含炎症和先天免疫基因的骨髓细胞相关，可预测不良反应我计划跟进这项工作并剖析先天免疫在两项 ICB 临床试验中的结果和生存率。膀胱癌的概况并研究骨髓细胞介导的 ICB 治疗耐药机制。目标 1 旨在定义未经治疗的膀胱肿瘤的情况并提供对免疫细胞的深入了解我将构建膀胱癌的转录组和分子图谱。单细胞分辨率，目前尚不存在的资源，我将建立患者的肿瘤、血液和细胞图谱。使用单细胞 RNA 测序、通过测序对转录组和表位进行细胞索引来检测尿液 (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 的软件对其进行分析我计划解析表达这种耐药基因特征的骨髓细胞，并定义它们的细胞相互作用。在目标 2 中，我将深入研究骨髓细胞中导致 ICB 耐药的转录途径。已经确定了肿瘤单核巨噬细胞（单核细胞）中的 NLRP3 炎性体激活和 IL-1β 信号传导 MΦs）作为促进肿瘤炎症和进展途径的候选者，我将通过以下方式对这些单 MΦs 进行建模。在低氧条件下使用 GM-CSF 和 M-CSF 使用 IL-将外周血单核细胞分化为 MΦ 1β和NLRP3炎性体激活剂。，我将通过共培养这些单-来测试对适应性免疫的影响我还将使用该模型来测试已知可调节的候选药物。 IL-1β 和 NLRP3 炎性体活性作为膀胱癌中 ICB 的潜在联合治疗。该提案将直接离体单细胞基因组学与体外功能实验相结合，以进行彻底的研究探究先天免疫对膀胱癌 ICB 耐药性的贡献，这些目标将结合起来。阐明先天免疫途径驱动的膀胱癌对 PD-1/PD-L1 ICB 治疗的耐药性，这可以用于识别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。