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: 10537272
• 负责人: Michelle Alyssa Tran
• 金额: $ 4.66万
• 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/10537272
• 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 Colony-Stimulating Factor; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mediating; Mediator of activation protein; 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; Signal Transduction; Specimen; System; T-Lymphocyte; Testing; Tumor Immunity; United States; Urine; Work; adaptive immune response; adaptive immunity; angiogenesis; anti-PD1 antibodies; base; 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; inhibitor; innate immune mechanisms; innate immune pathways; insight; macrophage; malignant breast neoplasm; marenostrin; monocyte; novel strategies; patient subsets; peripheral blood; predictive marker; predictive signature; resistance gene; resistance mechanism; response; 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.

项目总结