Role of ATRX, a chromatin remodeler, in immunotherapy response

ATRX（染色质重塑剂）在免疫治疗反应中的作用

基本信息

批准号：
10622315
负责人：
Daniel SANGHOON Shin
金额：
--
依托单位：
VA GREATER LOS ANGELES HEALTHCARE SYSTEM
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：
2022-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10622315
关键词：
ATAC-seq ATRX gene Acceleration Advanced Malignant Neoplasm Atlases Binding Biological Assay C57BL/6 Mouse CRISPR/Cas technology Cancer Model Cancer cell line Cell Line Cells ChIP-seq Chromatin Chromatin Structure Clinical Coculture Techniques Colorectal Cancer Complex Coupled Data Development Diagnosis EZH2 gene Elements Embryo Epigenetic Process Exhibits Feedback Fibroblasts Genes Genetic Transcription Genetically Engineered Mouse Genome Genomics Granzyme Health Histone Deacetylase Inhibitor Human IRF1 gene Immune Immune Evasion Immune system Immunocompetent Immunooncology Immunotherapy Interferon Receptor Interferon Type II Interferons JAK1 gene JAK2 gene Janus kinase Kidney Knock-out Lentivirus Vector MC38 Malignant Neoplasms Malignant neoplasm of lung Mediating Melanoma Cell Military Personnel Modeling Morbidity - disease rate Mus Mutation Outcome PD-1 blockade Parents Pathway interactions Patients Phenotype Play Polycomb Process Proteins RNA Sequences Reporting Research Research Design Research Proposals Resistance Rest Role Signal Pathway Signal Transduction Signaling Molecule Sleeping Beauty Small Interfering RNA Specificity System T-Lymphocyte Testing The Cancer Genome Atlas Therapeutic Transcript Transposase Treatment Efficacy Untranslated RNA Veterans XCL1 gene anti-PD1 antibodies anti-PD1 therapy antitumor effect cBioPortal cancer cell cancer immunotherapy chromatin remodeling demethylation design experimental study immune checkpoint blockade improved in vivo inhibitor loss of function loss of function mutation lung cancer cell melanoma mouse model mutant patient subsets perforin polybromo programmed cell death ligand 1 promoter receptor resistance mechanism response screening side effect tool transcriptome transcriptome sequencing treatment response trend tumor tumor growth vector

项目摘要

Background: Cancer immunotherapy is a major breakthrough for many patients with advanced cancer. However, benefits are still limited to a subset of patients, and we need to better understand the mechanisms of response and resistance to improve therapeutic efficacy. We have identified loss of function (LoF) mutations in JAK1 or JAK2 (immediate downstream signaling molecule of interferon receptor) that are associated with resistance to PD-1 blockade. We also found these mutations in human melanoma cell lines by screening PD- L1 expression with IFN-g treatment (48 cell lines). Tumors harbor LoF in JAK1/2, completely lost PD-L1 expression. Interestingly, one of them harbors no mutation with an active signaling pathway, yet lost PD-L1 expression. We explored why some human cancer cells lost adaptive PD-L1 expression even with intact interferon signaling and hypothesized that the epigenetic perturbation is mediating this phenotype. With this approach, we observed reduced PD-L1 expression with ATRX siRNA which was further tested with in vivo mouse models. In vivo mouse experiments with ATRX KO MC38 cells, anti-PD-1 antibody therapy produced either accelerated tumor growth or no effect. The current study is designed to understand the mechanism of resistance mediated by loss of ATRX in cancer immunotherapy. Objective/hypothesis: ATRX, a SWI/SNF-like chromatin remodeler is modulating the accessibility of interferon responsive genes that are associated with immunotherapy response. Specific aims: I have two aims for this study. The first aim is to interrogate the mechanisms of immune evasion with loss of ATRX using various tools to probe epigenetic state. The second aim is to establish in vivo tumor growth with ATRX KO using various murine cancer models. Study design: Aim 1. Subaim1) Generate ATRX KO B16 cells followed by Assess epigenetics state with IFN-g stimulation (both MC38 and B16 ATRX KO clones) using ChIP/ATAC-seq. Subaim 2) Correlate genomic studies (ChIP/ATAC) with Chromatin-Associated RNA-sequence (ChAR). Subaim 3) Assess the impact of epigenetic modifiers in IFN-g response in ATRX KO clones (using various epigenetic modifiers, such as HDAC inhibitor, demethylating agents and EZH2 inhibitor). Subaim 4) Coculture assay with murine T cells with ATRX wild-type parent cells and KO clones. Aim 2. Subaim 1) In vivo experiments with MC38 and B16 models with ATRX KO. Subaim2) Kras mutant murine lung cancer cell line models with ATRX KO. Subaim 3) ATRX KO in lung cancer and melanoma genetically engineered mouse models using sleeping beauty transposase vector system. Relevant to Military health: Improving treatment of many types of advanced cancers is critically important to the health of Veterans. Many Veterans suffer from significant morbidity when they are diagnosed with cancer that limits their therapeutic options. Immunotherapy is generally well tolerated and has a significant potential for durable response, however, the benefit is limited to a subset of patients (and Veterans). Therefore, it is imperative to improve therapeutic efficacy of immunotherapy by understanding the mechanisms of response and resistance. The proposed research is designed to understand the mechanisms of how cancer cells evade the immune system by modulating the chromatin state, focusing on the role of ATRX.

背景：癌症免疫疗法对于许多晚期癌症患者来说是一项重大突破。然而，获益仍然仅限于一小部分患者，我们需要更好地了解其机制反应和抵抗，以提高治疗效果。我们已经发现了功能丧失（LoF）突变 JAK1 或 JAK2（干扰素受体的直接下游信号分子）与对 PD-1 阻断具有抵抗力。我们还通过筛选 PD-在人类黑色素瘤细胞系中发现了这些突变 IFN-g 处理后的 L1 表达（48 个细胞系）。肿瘤在 JAK1/2 中含有 LoF，PD-L1 完全丢失表达。有趣的是，其中一个不存在具有活跃信号通路的突变，但丢失了 PD-L1 表达。我们探讨了为什么一些人类癌细胞即使在完整的 PD-L1 表达的情况下也失去了适应性 PD-L1 表达。干扰素信号传导并假设表观遗传扰动正在介导这种表型。有了这个方法中，我们观察到 ATRX siRNA 降低了 PD-L1 表达，并进一步进行了体内测试鼠标模型。 ATRX KO MC38细胞体内小鼠实验，产生抗PD-1抗体疗法要么加速肿瘤生长，要么没有效果。目前的研究旨在了解其机制癌症免疫治疗中 ATRX 缺失介导的耐药性。目标/假设：ATRX，一种类似 SWI/SNF 的染色质重塑剂正在调节与免疫治疗反应相关的干扰素反应基因。具体目标：我这项研究有两个目标。第一个目标是探究免疫机制使用各种工具探测表观遗传状态，通过丢失 ATRX 来逃避。第二个目标是建立体内使用各种小鼠癌症模型通过 ATRX KO 进行肿瘤生长。研究设计：目标 1. Subaim1) 生成 ATRX KO B16 细胞，然后使用 IFN-g 评估表观遗传学状态使用 ChIP/ATAC-seq 进行刺激（MC38 和 B16 ATRX KO 克隆）。 Subaim 2) 关联基因组染色质相关 RNA 序列 (ChAR) 的研究 (ChIP/ATAC)。 Subaim 3) 评估影响 ATRX KO 克隆中 IFN-g 反应的表观遗传修饰剂（使用各种表观遗传修饰剂，例如 HDAC 抑制剂、去甲基化剂和 EZH2 抑制剂）。 Subaim 4) 使用 ATRX 与小鼠 T 细胞共培养测定野生型亲代细胞和 KO 克隆。目标 2. Subaim 1) 使用 ATRX KO 进行 MC38 和 B16 模型的体内实验。 Subaim2) Kras 突变体 ATRX KO 小鼠肺癌细胞系模型。 Subaim 3) ATRX KO 在肺癌和黑色素瘤中的应用使用睡美人转座酶载体系统的基因工程小鼠模型。与军事健康相关：改善多种晚期癌症的治疗对于军事健康至关重要退伍军人的健康。许多退伍军人在被诊断患有癌症时患有严重的疾病这限制了他们的治疗选择。免疫疗法通常具有良好的耐受性，并且具有巨大的潜力然而，持久的反应，其益处仅限于一小部分患者（和退伍军人）。因此，它是通过了解反应机制来提高免疫疗法的疗效势在必行和阻力。拟议的研究旨在了解癌细胞如何逃避的机制免疫系统通过调节染色质状态，重点关注ATRX的作用。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Enhanced CTLA-4 blockade anti-tumor immunity with APG-157 combination in a murine head and neck cancer.

在小鼠头颈癌中联合 APG-157 增强 CTLA-4 阻断抗肿瘤免疫力。

DOI：
10.1002/cam4.7212
发表时间：
2024-04-30
期刊：
Cancer medicine
影响因子：
4
作者：
Daniel Sanghoon Shin;S. Basak;M. Veena;B. Comin;Arjun Bhattacharya;Tien S Dong;Albert Y Ko;Philip Han;Jonathan Jacobs;Neda A Moatamed;Luis Avila;Matteo Pellegrini;Marilene B Wang;E. Srivatsan
通讯作者：
E. Srivatsan