Effects of entinostat and neoantigen vaccination on bladder cancer

恩替司他和新抗原疫苗接种对膀胱癌的影响

基本信息

批准号：
10751492
负责人：
Wolfgang Beckabir
金额：
$ 4.61万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
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/10751492
关键词：
Affect Antigens Automobile Driving Bioinformatics CD8-Positive T-Lymphocytes Cancer Model Cancer Patient Caring Cessation of life Clinical Trials Collaborations Computer software Development Epigenetic Process Flow Cytometry Funding Future Genetic Genitourinary system Genomics Growth HDAC1 gene Histone Deacetylase Inhibitor Human Immune checkpoint inhibitor Immunofluorescence Immunologic Immunologics Immunology In Vitro Knowledge Malignant Neoplasms Malignant neoplasm of urinary bladder Measures Migration Assay Modeling Mus Oncology Patients Physicians Population Positioning Attribute Prognosis Regulatory T-Lymphocyte Research Sampling Scientist Single Nucleotide Polymorphism Source Specificity Stains Statistical Models Survival Rate T cell response T-Cell Immunologic Specificity T-Lymphocyte Testing Training Translational Research Tumor Antigens Tumor Biology Tumor Volume United States Work anti-PD-1 cancer diagnosis cancer immunobiology career effective therapy enzyme linked immunospot assay experience immune checkpoint blockade immunogenic immunogenicity improved in vivo men migration mouse model neoantigen vaccination neoantigens response single-cell RNA sequencing subcutaneous transcriptome sequencing transcriptomics treatment effect tumor tumor immunology tumor microenvironment ultrasound

项目摘要

Project Summary/Abstract Bladder cancer is a prevalent and deadly cancer, with over 80,000 new cases and 17,000 deaths annually in the United States. Advanced bladder cancer has only a 15% 5-year survival rate. One of the most effective treatments for advanced bladder cancer is immune checkpoint blockade (ICB), but only 20-30% of patients with advanced bladder cancer respond and most responses are not enduring. A promising new treatment identified by our group to improve bladder cancer ICB response is the selective class 1 histone deacetylase inhibitor entinostat. In a mouse model, entinostat plus anti-PD-1 (αPD-1) ICB induced complete, enduring responses in 67% of mice. Entinostat decreased intratumoral M-MDSC and Treg populations, decreased tumor single- nucleotide variant (SNV) neoantigen burden in vivo, increased expression of some SNV neoantigens in vitro, and increased T cell specificity for these neoantigens in vivo. However, much of the mechanism behind response to entinostat plus αPD-1 is unknown, particularly how entinostat decreases immunosuppressive populations and affects expression of the neoantigen landscape. Understanding this mechanism is important to predict which patients will respond and to potentially improve responses through antigen-directed therapy. We hypothesize that entinostat-induced ICB response is driven by increased M-MDSC differentiation, decreased M-MDSC migration, and increased expression of suppressed immunogenic neoantigens, augmenting response to neoantigen vaccination. I will investigate two components driving response to entinostat plus αPD-1: M-MDSCs and neoantigens. The training in computational and wet lab immunology, tumor biology and genetics, orthotopic murine tumor models, and translational research with a clinical trial, will assist me in becoming an independently funded physician-scientist leading a cancer immunology research lab and caring for bladder cancer patients. In our M-MDSC-focused Aim 1, I will perform flow cytometry and transwell migration assays with M-MDSCs from orthotopic bladder cancer model tumors to assess whether M-MDSC differentiation and migration are affected by entinostat treatment. I will conduct immunofluorescence staining of human tumors from the LCCC1827 entinostat window trial (NCT03978624) to assess whether adding entinostat to ICB treatment decreases M- MDSCs in humans. The Vincent Lab has developed LENS, a software platform to identify neoantigens from multiple genomic sources. In our neoantigen expression-focused Aim 2, I will use LENS to identify all the neoantigens in 3 murine bladder cancer lines developed by the Kim Lab, test T cell neoantigen specificity by high-throughput ELISPOT, and use statistical modeling to predict neoantigens immunogenicity. I will also validate whether entinostat-induced immunoediting occurs in human tumors from LCCC1827. In our neoantigen vaccination-focused Aim 3, I will test whether neoantigen vaccination improves tumor response to entinostat plus αPD-1. I will treat orthotopic tumors, measure their growth, and perform flow cytometry and single cell RNAseq to measure non-exhausted neoantigen-specific CD8+ T cell abundance.

项目概要/摘要膀胱癌是一种流行且致命的癌症，每年有超过 80,000 例新发病例和 17,000 例死亡美国晚期膀胱癌的 5 年生存率仅为 15%，是最有效的治疗方法之一。晚期膀胱癌的治疗方法是免疫检查点阻断 (ICB)，但只有 20-30% 的患者晚期膀胱癌有反应，但大多数反应并不持久。我们小组改善膀胱癌的 ICB 反应是选择性 1 类组蛋白脱乙酰酶抑制剂在小鼠模型中，恩替司他加抗 PD-1 (αPD-1) ICB 可诱导完全、持久的反应。 67% 的小鼠恩替司他减少了肿瘤内 M-MDSC 和 Treg 群体，减少了肿瘤单细胞。体内核苷酸变异（SNV）新抗原负荷，体外某些SNV新抗原表达增加，并增加了体内这些新抗原的 T 细胞特异性。然而，反应背后的大部分机制。恩替司他加 αPD-1 的作用尚不清楚，特别是恩替司他如何减少免疫抑制群体和影响新抗原景观的表达对于预测哪种机制很重要。患者将做出反应，并可能通过抗原导向治疗改善反应。恩替司他诱导的 ICB 反应是由 M-MDSC 分化增加、M-MDSC 减少驱动的迁移，并增加受抑制的免疫原性新抗原的表达，增强对我将研究驱动恩替司他加 αPD-1 反应的两个成分：M-MDSC。计算和湿实验室免疫学、肿瘤生物学和遗传学、原位培训。小鼠肿瘤模型以及临床试验的转化研究将帮助我成为一名独立的资助的医师科学家领导癌症免疫学研究实验室并照顾膀胱癌患者。在我们以 M-MDSC 为重点的目标 1 中，我将使用来自原位膀胱癌模型肿瘤评估 M-MDSC 分化和迁移是否受到影响我将对 LCCC1827 的人类肿瘤进行免疫荧光染色。恩替司他窗口试验 (NCT03978624) 评估在 ICB 治疗中添加恩替司他是否会降低 M- Vincent 实验室开发了 LENS，这是一个用于识别人类 MDSC 中新抗原的软件平台。在我们以新抗原表达为重点的目标 2 中，我将使用 LENS 来识别所有基因组来源。 Kim 实验室开发的 3 种小鼠膀胱癌细胞系中的新抗原，通过以下方法测试 T 细胞新抗原特异性：我还将使用高通量 ELISPOT 并使用统计模型来预测新抗原的免疫原性。在我们的新抗原中验证恩替司他诱导的免疫编辑是否发生在人类肿瘤中。以疫苗接种为重点的目标 3，我将测试新抗原疫苗接种是否可以改善肿瘤对恩替司他的反应加上 αPD-1，我将治疗原位肿瘤，测量其生长，并进行流式细胞术和单细胞分析。 RNAseq 用于测量未耗尽的新抗原特异性 CD8+ T 细胞丰度。