Malignant Cell Engagement in Immune Circuits of Human Microsatellite-stable and Microsatellite-instable Colorectal Cancer

恶性细胞参与人类微卫星稳定和微卫星不稳定结直肠癌的免疫回路

• 批准号: 10469715
• 负责人: Karin Pelka
• 金额: $ 24.9万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-08 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10469715
• 关键词: Malignant; Cell; Engagement; Immune; Circuits

Project Summary An important question in cancer immunology is how malignant cells and T cells communicate and impact the efficacy of anti-tumor immunity. To generate hypotheses for how these cells interact, we have applied single cell RNA sequencing to profile the transcriptomes of >700,000 malignant, immune, and stromal cells in 2 cohorts: (1) 65 patients with two types of primary untreated CRC (31 MSS and 34 MSI tumors) which are characterized by vastly different immunotherapy response rates; (2) pre- and post-treatment specimens from 20 BRAF-mutant metastatic CRC patients treated with BRAFi/MEKi/anti-PD1. These data led us to formulate the central hypothesis for this proposal: malignant cells that express interferon-stimulated genes (ISGs) acutely promote anti-tumor immunity, while chronic interferon responses in malignant cells impair anti-tumor immunity and therapeutic responses. The proposed work is anticipated to reveal fundamental insights into human tumor immunology and provide a novel perspective for the design of robust biomarkers and new therapeutic strategies. We will address three major questions. First, is the MSI/responder-associated ISG program in malignant cells part of an intra-tumoral feed-forward loop that is driving anti-tumor immunity? This will be answered by spatially mapping the intratumoral cellular interaction network between ISG+ malignant cells and T cells in untreated primary CRC specimens, and in post-treatment specimens of patients treated with BRAFi/MEKi/anti-PD-1, which will reveal association of this spatial network with tumor regression. Second, what are the drivers of malignant ISGs in human CRC, and can interferons impact immune responses and responses to targeted therapy by epigenetically reprogramming human CRC cells? To address this, we will use interferons and innate immune stimuli to induce ISGs in CRC organoids, map the transcriptional and epigenetic signatures to the signatures found in freshly isolated CRC cells, and nominate ISG-inducing upstream regulators. IFN-stimulated and subsequently rested organoids will be assessed for epigenetic memory and for modulated secondary immune and drug responses. Third, how do pre-existing ISG signatures in malignant cells impact CTL-mediated anti-tumor responses? Using peptide-cognate T cell lines, we will test the in vitro killing of ISG+ and ISG- peptide-loaded CRC organoids. We will furthermore test how ISGs modulate in vivo tumor killing by temporally controlling malignant ISG expression in transplantable tumor models. In summary, we use scRNAseq to predict malignant cell – T cell interactions, and then test these hypotheses with cutting-edge technologies such as spatial profiling, single cell epigenomics, and human organoid T cell co-cultures. The results should elucidate how malignant cells that respond to interferons impact anti-tumor immunity and resistance, generate foundational results and model systems for an R01 proposal, and prepare the candidate for an independent career to study the cellular networks that drive human tumor immunity.

项目摘要 癌症无限学中的一个重要问题是，恶性细胞和T如何交流和影响 抗肿瘤免疫的功效。 细胞RNA测序以介绍2个> 700,000个恶性，免疫和基质细胞的转录。 队列：（1）65例患有两种类型的原发性CRC（31 MS和34个MSI肿瘤）的患者是 以截然不同的反反应率为特征； 20个用BRAFI/MEKI/抗PD1治疗的BRAF突变转移性CRC患者。 该弹性的中心假设：表达干扰素刺激基因（ISGS）敏捷的恶性细胞 促进抗肿瘤免疫，而恶性细胞中的慢性干扰素反应会损害抗肿瘤IMUNITY 和治疗的反应。 免疫学并为鲁棒生物标志物设计和新的治疗性设计提供了一种新颖的观点 策略。 恶性细胞的一部分是朝向前循环的一部分，该环路正在驱动抗肿瘤的进口？ 通过空间绘制ISG+恶性细胞和 未经治疗的原代CRC标本中的T细胞，以及与接受治疗的患者的治疗后样品中的T细胞。 BRAFI/MEKI/抗PD-1，该空间网络将与肿瘤退化的关联。 人CRC中恶性ISG的驱动因素是什么，可以干扰免疫反应和 我们会通过表观遗传重新编程的人CRC细胞对靶向治疗的反应？ 使用干扰素和先天的免疫刺激在CRC器官中诱导ISGS，绘制转录和 在新鲜分离的CRC细胞中发现的特征的表观遗传学特征，并提名ISG诱导 上游调节剂。 记忆和模块化的次级免疫和药物反应。 在恶性细胞中，CTL介导的抗肿瘤反应？ ISG+和ISG肽的体外杀死CRC器官。 通过暂时控制可移植肿瘤中的恶性ISG表达来杀死体内肿瘤的模块化酸盐 总而言 带有尖端技术的假设，例如空间分析，单细胞表观基因组学和人类 有机体T细胞共培养。结果应阐明对干扰素响应的恶性细胞 抗肿瘤的免疫力和抵抗力，为R01提案生成基础结果和模型系统， 并准备独立职业的候选人研究驱动人肿瘤的细胞网络 免疫。