Deeply analyzing MHC class I-restricted peptide presentation mechanistics across alleles, pathways, and disease coupled with TCR discovery/characterization

深入分析跨等位基因、通路和疾病的 MHC I 类限制性肽呈递机制以及 TCR 发现/表征

• 批准号: 10674405
• 负责人: Roland K Strong
• 金额: $ 80.42万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674405
• 关键词: Address; Adoptive Immunotherapy; Alleles; Antibodies; Antigens; Award; Basic Science; Binding; Biochemical; Biological Assay; Cancer Patient; Cancerous; Cell surface; Cells; Cellular Assay; Characteristics; Clinical; Collaborations; Complex; Coupled; Crystallography; Data; Diagnosis; Disease; Epitopes; Future; HLA G antigen; HPV-High Risk; Histocompatibility Antigens Class I; Human; Human Papillomavirus; Human papillomavirus 16; Immune system; Immunotherapy; Individual; Infection; MHC Class I Genes; Malignant Neoplasms; Maps; Mesothelioma; Methodology; Microbe; Modality; Modeling; Molecular; Molecular Conformation; Monitor; Non-Small-Cell Lung Carcinoma; Oncoproteins; Papillomavirus Transforming Protein E6; Pathway interactions; Patients; Peptide Fragments; Peptide/MHC Complex; Peptides; Pleural effusion disorder; Process; Proteins; Proteome; Quality Control; Renal Cell Carcinoma; Research; Sampling; Science; Specificity; Stains; Surface; System; T cell response; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; Time; Tumor Antigens; Tumor-Infiltrating Lymphocytes; Vaccine Therapy; Viral; antigen processing; cancer immunotherapy; clinical application; comparative; engineered T cells; experimental study; glycosylation; immunogenicity; improved; mesothelin; multicatalytic endopeptidase complex; new therapeutic target; novel; pathogen; prostatic fraction Acid phosphatase isoenzyme; receptor; response; single cell sequencing; tumor

Project Abstract/Summary • Our framing hypothesis is that the combined constraints of peptide processing, HLA allelic specificity, and peptide editing generate a significant “funnel effect”, where the peptidome constitutes only a tiny percentage of all possible peptides obtainable from a cell’s proteome and the ligandome constitutes only a small percentage of all peptides from a cell’s peptidome. Our research focus is understanding how the interplay of differential sequence recognition at each step ultimately informs functional presentation of a highly restricted subset of all possible proteome-derived peptides – and how to identify the most “translatable” pHLA targets from ligandomes. • We will rigorously test this hypothesis using our allele-specific mass-spec platform for peptide discovery, ARTEMIS, to identify relevant, HLA-restricted peptides from Mesothelin and high-risk HPV E6/E7 oncoproteins. Identified peptides will be validated and characterized biochemically. Cell-surface displayed peptide arrays from E6/E7 and Mesothelin will be used to determine the magnitude of the “funnel effect” of peptide processing on peptidome composition. Patient T cell responses to identified Mesothelin peptides will be mapped using state- of-the-art methodologies. Unusual peptides and alternate binding conformations will be characterized crystallo- graphically. We will assess viral immunoevasion mechanisms, HLA-G presentation and receptor interactions, and the presentation of glycosylated peptides. Deliverables include mapping the full Mesothelin and E6/E7 presentomes across multiple alleles and presentation modalities, ranked for future clinical exploitation as immu- notherapy targets. • Significance: From a basic science perspective, assaying cellular peptidomes and ligandomes is fundamental for understanding how the immune system responds to infections and cancer, immunoevasion mechanisms, the recognition rules of MHC proteins, the mechanistics of antigen processing/editing, and how self is defined. From a translational perspective, validated HLA-restricted peptides represent targets for diagnosis and therapy, through therapeutic vaccination, engineered T cell-based adoptive immunotherapies, and antibody-based recog- nition of specific, disease-associated peptide/HLA complexes Our proposed studies advance both objectives by elucidating the underlying molecular principles governing peptide processing and presentation through inte- grated studies of an extended set of classical and non-classical human HLA class I proteins and, in the process, identify specific pHLAs useful for future clinical applications.

项目摘要/摘要 ·我们的框架假设是肽加工、HLA等位基因特异性和 肽编辑产生显著的"漏斗效应"，其中肽组仅占 从细胞蛋白质组和配体组获得的所有可能的肽仅占很小的百分比 细胞肽组中所有肽的信息。我们的研究重点是了解差异的相互作用 在每一步的序列识别最终通知所有的高度限制的子集的功能呈现。 可能的蛋白质组衍生肽-以及如何从配体组中识别最"可翻译"的pHLA靶点。 ·我们将使用我们用于肽发现的等位基因特异性质谱平台严格测试这一假设， ARTEMIS，以识别来自间皮素和高危HPV E6/E7癌蛋白的相关HLA限制性肽。 将对识别的肽进行生物化学验证和表征。细胞表面展示肽阵列， 将使用E6/E7和间皮素来确定肽加工对细胞的"漏斗效应"的大小。 肽组组成。患者T细胞对所鉴定的间皮素肽的应答将使用状态- 最先进的方法不寻常的肽和替代结合构象将被表征晶体- 图形地。我们将评估病毒免疫逃避机制，HLA-G呈递和受体相互作用， 和糖基化肽的呈递。可交付成果包括绘制完整的间皮素和E6/E7 presentomes跨越多个等位基因和呈递方式，被列为未来临床开发的免疫- notherapy目标 ·意义：从基础科学的角度来看，测定细胞肽组和配体组是基础 为了了解免疫系统如何对感染和癌症作出反应，免疫逃避机制， MHC蛋白的识别规则，抗原加工/编辑的机制，以及自我是如何定义的。从 从翻译的角度来看，经验证的HLA限制性肽代表了诊断和治疗的靶点， 通过治疗性疫苗接种、基于工程化T细胞的过继免疫疗法和基于抗体的识别， 特异性疾病相关肽/HLA复合物的识别我们提出的研究通过以下方式推进了这两个目标： 阐明了控制肽加工和呈递的基本分子原理， 对经典和非经典人类HLA I类蛋白的扩展集的分级研究，在此过程中， 鉴定对未来临床应用有用的特异性pHLA。