Investigating splicing-derived antigens in the context of Rbm10-mutant lung adenocarcinoma

研究 Rbm10 突变肺腺癌中的剪接衍生抗原

基本信息

批准号：
10606839
负责人：
Nicolas Mathey-Andrews
金额：
$ 5.27万
依托单位：
HARVARD MEDICAL SCHOOL
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：
2023-09-01 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10606839
关键词：
Adaptive Immune System Adenocarcinoma Advanced Malignant Neoplasm Animals Antibodies Antigen Presentation Antigens Binding Bioinformatics Biological Markers CD8-Positive T-Lymphocytes CD8B1 gene CRISPR/Cas technology Cancer Etiology Cells Cessation of life Common Neoplasm Development Dose Epitopes Event Exhibits Genes Genetically Engineered Mouse Genome engineering Genotype Goals Growth Histologic Human Immune Immune response Immune system Immunocompetent Immunoprecipitation Immunotherapy Infiltration Knock-out Lung Lung Adenocarcinoma Lung Neoplasms Lymphocyte Malignant Neoplasms Malignant neoplasm of lung Mass Spectrum Analysis Mediating Methods Modeling Mutation Patients Pattern Peptide/MHC Complex Peptides Predisposition Productivity Protein Fragment Proteins Publishing RNA Splicing Recurrence Role Somatic Mutation Suppressor Mutations T cell response T-Cell Depletion T-Lymphocyte Testing Transcript Translating Translations Tumor Antigens Tumor Burden Tumor Immunity Work adaptive immune response anti-PD-1 anti-tumor immune response antigen-specific T cells cancer cell cancer genome cancer immunotherapy clinical predictors cohort driver mutation immune cell infiltrate immune checkpoint blockade immunogenic immunogenicity in vivo Model innovation loss of function mutation mRNA sequencing mouse model mutant neoantigens neoplastic cell novel novel therapeutics patient stratification patient subsets predicting response programmed cell death ligand 1 research study response restraint standard of care transcriptome sequencing tumor

项目摘要

Project summary: The adaptive immune system can specifically recognize and kill cancer cells. Therapies that reinvigorate tumor-specific CD8 T cells cure a subset of patients with lung cancer, prompting their FDA approval and use as standard of care. Tumor-reactive T cells recognize cancer cell antigens -- displayed fragments of proteins that are unique to tumor cells. To date, study of anti-tumor immunity has primarily focused on antigens derived from somatic mutations in the cancer genome, which are insufficient to fully explain the anti-tumor immune response. Moreover, mutation-derived neoantigens are generally unique to each tumor, and targeting them might require bespoke treatments for each patient. The long-term goal of this proposal is to expand the paradigm of cancer-specific antigens. Recent work has uncovered a novel class of cancer-restricted antigens produced by abnormal splicing events. While several cancers harbor recurrent driver mutations in splicing factors, few studies have explored the relationship between dysregulated splicing and tumor immunogenicity. Notably, ~10% of lung adenocarcinomas feature loss-of-function mutations in RBM10, a critical splicing factor. This work will examine splicing-derived antigens in Rbm10-deficient tumors. The central hypothesis is that Rbm10 deficiency will generate aberrantly spliced transcripts, which will elicit an adaptive immune response if translated and presented as novel antigens by tumor cells. This hypothesis will be tested in a well- characterized murine model of lung adenocarcinoma by pursuing two specific aims. In Aim 1, mRNA sequencing will be performed to characterize aberrant splicing events that occur in the context of Rbm10 deficiency. Mass spectrometry will be used to validate that aberrantly spliced RNAs produce antigens presented to the immune systems. In Aim 2, CRISPR/Cas9 will be used to introduce Rbm10 mutations in an immunocompetent, autochthonous murine model. Histologic and flow cytometric analyses will be performed on infiltrating lymphocytes in Rbm10-deficient and proficient tumors to assess endogenous anti-tumor immunity. This research study is innovative in that it proposes to empirically test the role of splicing-derived antigens in anti-tumor immunity using genome engineering in autochthonous mouse models. It is significant because it may identify new, tumor-restricted antigens that are targetable by cancer immunotherapies. While T-cell targeted immunotherapies provide durable cures for patients with advanced cancer, prior studies emphasize a limited set of cancer-specific T cell antigens. The ultimate objective of this work is to uncover novel mechanisms to potentiate immune recognition of cancer cells, leading to the development of new therapies for lung cancer.

项目概要：适应性免疫系统可以特异性识别并杀死癌细胞。恢复活力的疗法肿瘤特异性 CD8 T 细胞治愈了一部分肺癌患者，促使 FDA 批准和使用它们作为护理标准。肿瘤反应性 T 细胞识别癌细胞抗原——展示的蛋白质片段是肿瘤细胞特有的。迄今为止，抗肿瘤免疫的研究主要集中在抗原来源来自癌症基因组中的体细胞突变，这不足以完全解释抗肿瘤免疫回复。此外，突变衍生的新抗原通常对每种肿瘤都是独特的，并且针对它们可能需要为每位患者定制治疗。该提案的长期目标是扩大癌症特异性抗原的范例。最近的工作发现了一类新型癌症限制抗原由异常剪接事件产生。虽然几种癌症在剪接中存在反复的驱动突变因素，很少有研究探讨剪接失调与肿瘤免疫原性之间的关系。值得注意的是，大约 10% 的肺腺癌具有 RBM10（一种关键剪接因子）的功能丧失突变。这项工作将检查 Rbm10 缺陷肿瘤中剪接衍生的抗原。中心假设是 Rbm10 缺陷会产生异常剪接转录本，从而引发适应性免疫如果肿瘤细胞翻译并呈递新抗原，则会产生反应。这个假设将在一个良好的环境中得到检验通过追求两个特定目标来表征肺腺癌的小鼠模型。在目标 1 中，mRNA 将进行测序以表征 Rbm10 背景下发生的异常剪接事件不足。质谱法将用于验证异常剪接的 RNA 是否会产生抗原呈现给免疫系统。在目标 2 中，CRISPR/Cas9 将用于在细胞中引入 Rbm10 突变。具有免疫功能的本土小鼠模型。将进行组织学和流式细胞术分析在 Rbm10 缺乏和丰富的肿瘤中浸润淋巴细胞，以评估内源性抗肿瘤免疫。这项研究的创新之处在于，它提出以实证方式测试剪接衍生抗原在在本地小鼠模型中使用基因组工程的抗肿瘤免疫。这很重要，因为它可能会识别出癌症免疫疗法可靶向的新的肿瘤限制性抗原。而T细胞靶向免疫疗法为晚期癌症患者提供持久治愈，先前的研究强调有限的癌症特异性 T 细胞抗原。这项工作的最终目标是发现新颖的增强癌细胞免疫识别的机制，导致新疗法的开发肺癌。