"Post-translational modification of non-histone proteins as a mechanism of pMHC-I neo-ligand generation"

“非组蛋白蛋白的翻译后修饰作为 pMHC-I 新配体生成的机制”

• 批准号: 10435184
• 负责人: ROBERT K BRIGHT
• 金额: $ 7.65万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-03 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10435184
• 关键词: Acetylation; Address; Affect; Affinity; Amino Acids; Antibody Response; Antigens; Autoantigens; Autoimmune; Avidity; Behavior; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Vaccines; Cell model; Cells; Data; Deacetylase; Deacetylation; Development; Discrimination; Epitopes; Generations; Goals; Immune; Immune Targeting; Immunity; Immunotherapy; In Vitro; Incubated; Investigation; Knowledge; Letters; Life; Ligands; Lysine; Malignant - descriptor; Masks; Mass Spectrum Analysis; Metastatic Neoplasm to the Lung; Mission; Modeling; Modification; Monitor; Mus; Neoplasm Metastasis; Non-Malignant; Normal Cell; Oncogenic; Organism; Peptide/MHC Complex; Peptides; Play; Post-Translational Protein Processing; Proteins; Public Health; Regulation; Regulatory T-Lymphocyte; Reporting; Research; Role; T-Lymphocyte; TPD52 gene; Testing; Thymus Gland; Tissues; Toxic effect; Translations; Tumor Immunity; United States National Institutes of Health; Vaccinated; Vaccination; Vaccine Antigen; Vaccines; Work; blind; burden of illness; cancer cell; cancer immunotherapy; cancer prevention; cancer vaccination; clinical application; immune checkpoint blockade; improved; in vivo; in vivo Model; inhibitor; neoantigens; non-histone protein; novel; novel vaccines; overexpression; pre-clinical; prevent; tumor; tumor growth; tumor microenvironment; tumorigenesis; vaccine failure

Project Summary Abstract The advancement of cancer vaccines requires a deeper understanding of T cell recognition of malignant vs non-malignant cells. Though the T cell immune repertoire is developmentally regulated by negative and positive selection, T cells capable of recognizing self-proteins do exist and are critical effectors for many cancer immunotherapies. What remains unclear is how CD8+ CTLs elicited against tumor-self proteins are capable of recognizing malignant cells yet remain blind to healthy cells that also express the self-protein. Lack of clarity on this issue may inhibit the advancement of cancer vaccines that target non-mutated, oncogenic, tumor-self proteins. Our long-term goal is a deeper understanding of how tumor-self antigen vaccine elicited CTLs recognize cancer cells but do not recognize normal cells that also express the antigen. Our immediate objective, defining the first critical step toward our long-term goal, is to determine if a difference in malignant and non-malignant cell post-translation modification of a relevant, model tumor-self antigen generates pMHC-I- neo-ligand(s) recognized on cancer cells by vaccine elicited CTLs. We hypothesize that post-translational deacetylation of lysine residues within a relevant, model tumor-self protein naturally differs between malignant and non-malignant cells resulting in the generation of pMHC-I-neo-epitopes on malignant cells. Guided by supportive preliminary data, our hypothesis will be tested with two specific aims: 1) Determine if tumor-self antigen-lysine deacetylation in malignant cells enables recognition by tumor-self antigen vaccine elicited CTLs. Using mass spectroscopy, we expect to define differential acetylation of lysine residues in the relevant, model, oncogenic tumor-self protein D52 expressed in malignant and non-malignant cells. We expect to demonstrate that D52 lysine acetylation effects D52 vaccine elicited CTL recognition of malignant cells resulting in differential CTL discrimination of malignant and non-malignant cells. Using a specific panel of peptides derived from D52, we anticipate that we will define D52 vaccine elicited CTL pMHC-I-neo-epitopes. 2) Determine if tumor-self antigen-lysine deacetylation in malignant cells impacts vaccine induced protective tumor immunity. Using malignant cells incubated with deacetylase inhibitor prior to inoculation into D52 vaccinated mice with monitoring of tumor growth overtime and endpoint assessment of lung metastases, we expect to demonstrate that inhibition of natural deacetylation of D52 in malignant cells impacts pMHC-I- epitopes that are recognized by CTLs possibly resulting in failure of the vaccine to protect against tumor growth and metastases. Impact: The results of this proof of concept project are expected to positively impact the development of more effective next-generation vaccines that target oncogenic tumor-self proteins for the safe treatment or prevention of cancer. Equally important is a deeper understanding of immunotherapy elicited CTLs and malignant, non-malignant cell discrimination that may impact auto-immune-related toxicities as has been reported for immune checkpoint blockade, and may be associated with next-generation vaccines.

项目摘要摘要 癌症疫苗的进步需要更深入地了解T细胞对恶性肿瘤的识别 非恶性细胞。尽管T细胞免疫曲目在发育中受到负面调节，并且 积极选择，能够识别自蛋白的T细胞确实存在，并且是许多人的关键效应子 癌症免疫疗法。尚不清楚的是如何针对肿瘤自身蛋白引起CD8+ CTL 能够识别恶性细胞但对也表达自我蛋白质的健康细胞视而不见。缺少 关于这个问题的明确性可能会抑制靶向无突出，致癌性的癌症疫苗的进步 肿瘤自身蛋白。我们的长期目标是对肿瘤自我抗原疫苗的产生更深入的了解 CTL识别癌细胞，但不识别也表达抗原的正常细胞。我们的直接 目的，定义朝着我们的长期目标的第一步，是确定恶性的差异 相关，模型肿瘤抗原的翻译后的非恶性细胞会产生PMHC-I- 通过疫苗引起的CTL在癌细胞上识别的NEO-配体（S）。我们假设翻译后 相关模型肿瘤蛋白质中赖氨酸残基的脱乙酰化自然之间存在变性 和非恶性细胞，导致在恶性细胞上产生PMHC-I-NEO- epepopes。指导 支持性初步数据，我们的假设将以两个具体的目的进行检验：1）确定肿瘤自我是否 恶性细胞中的抗原 - 赖氨酸脱乙酰基化可以通过肿瘤自身抗原疫苗识别 引起CTL。使用质谱法，我们期望定义赖氨酸残基的差异乙酰化 相关的模型，在恶性细胞和非恶性细胞中表达的致癌肿瘤蛋白D52。我们期望 为了证明D52赖氨酸乙酰化作用D52疫苗引起CTL识别恶性细胞 导致恶性细胞和非恶性细胞的CTL区分差异。使用特定的面板 源自D52的肽，我们预计我们将定义D52疫苗引起的CTL PMHC-i-Neo-epitopes。 2） 确定恶性细胞中肿瘤自我抗原 - 赖氨酸脱乙酰基化是否会影响疫苗诱导的 保护性肿瘤免疫。在接种之前，使用与脱乙酰基酶抑制剂孵育的恶性细胞 D52通过监测肿瘤生长的加班和肺转移终点评估的接种小鼠， 我们预计会证明，恶性细胞中D52自然脱乙酰化的抑制会影响PMHC-II CTL识别的表位可能导致疫苗预防肿瘤的疾病 生长和转移。影响：该概念证明项目的结果有望积极影响 靶向致癌肿瘤蛋白的更有效的下一代疫苗的开发 安全治疗或预防癌症。同样重要的是对引起免疫疗法的更深入了解 CTL和恶性，非恶性细胞歧视可能会影响自动免疫相关的毒性 据报道，免疫检查点封锁，可能与下一代疫苗有关。