Investigation into the Endogenous Ligand Repertoire of the Non-Classical MHC-Related Protein, MR1 in Multiple Myeloma Cell Lines

多发性骨髓瘤细胞系中非经典 MHC 相关蛋白 MR1 内源配体库的研究

• 批准号: 10452305
• 负责人: Erin June Adams
• 金额: $ 23.86万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452305
• 关键词: Activities of Daily Living; Antigen Presentation; Antigens; Binding; Biochemical; Biochemistry; Biological Assay; Blood Circulation; Blood donor; Blood specimen; Cancer cell line; Cancerous; Catalogs; Cell Line; Cell surface; Cells; Chemical Structure; Chemicals; Clone Cells; Collaborations; Complex; Development; Disease; Folic Acid; Folic Acid Derivative; Future; Genetic Transcription; Genomics; Goals; Health Sciences; Human; Immune; Immunotherapy; Individual; Infection; Investigation; Lead; Ligand Binding; Ligands; Light; MHC antigen; Major Histocompatibility Complex; Malignant Neoplasms; Metabolic Pathway; Modeling; Molecular; Mucous Membrane; Multiple Myeloma; Natural Products; Nature; Oklahoma; Pathway interactions; Patients; Peptides; Phenotype; Plasma Cells; Play; Population; Positioning Attribute; Property; Proteins; Protocols documentation; Reagent; Recombinants; Recording of previous events; Reporting; Research; Riboflavin; Role; Stains; Surface; T-Cell Receptor Genes; T-Lymphocyte; T-cell receptor repertoire; TRANCE protein; Techniques; Testing; Tissues; Tumor Antigens; Universities; Ursidae Family; Work; adaptive immunity; alpha-beta T-Cell Receptor; base; biobank; cancer cell; cancer type; cytokine; cytotoxic; exhaustion; experimental study; innovative technologies; interest; metabolome; microbial; microorganism antigen; neoplastic cell; novel; pathogen; response; sensor; sequencing platform; small molecule; social; structural biology; tumor

Project Abstract: Major Histocompatibility Complex (MHC) molecules have been long been appreciated as the bridge between the innate and adaptive immune cell populations. Beyond the peptide-presenting classical MHC class I and II molecules, there exists a variety of other structurally related proteins in humans that have been found to present diverse antigen types to a variety of T cell sub-types. Among these is the monomorphic MHC-related protein 1 (MR1) which is historically known for the presentation of microbially-derived riboflavin metabolites to Mucosal- Associated Invariant T (MAIT) cells. Expanding on this initial observation, our group and others have shown that the ligandome of MR1 is much more diverse than previously appreciated. MR1 is capable of presenting ligands of various sizes and chemical structures beyond the originally described ribityl metabolites to a vast array of MR1-restricted T cells (MR1Ts) carrying fully rearranged αβ TCRs. Recently, there have been emerging reports of MR1T clones that have shown MR1-dependent cytotoxic activity against cancerous tissues in the complete absence of any microbial infection or microbial ligands. Due to the unprecedented nature of these observations and the potential impact of an MR1-based immunotherapy, it is important that the ligands being presented by MR1 in the context of cancer-induced cellular disfunction be identified. Taking into consideration our own research and these novel reports, we propose that MR1 acts as a sensor of the cellular metabolome in both the context of microbial infection and endogenous disfunction like cancer, presenting a diverse array of metabolite antigens. Due to our established track record investigating the biochemistry of MR1 and our existing collaborative ligand discovery pipeline for MR1, The Adams Lab is uniquely poised to investigate the identity of the antigens that MR1 is presenting in the context of cancer. Our investigation will focus on the plasma cell cancer, multiple myeloma, as cell lines of this cancer have been shown to have abnormally high expression of MR1 at the cell surface, indicating a higher possible antigenic load. We propose that our established microbial-ligand discovery pipeline for MR1 can be adapted to investigate the tumor associated antigens (TAAs) being presented on the surface of myeloma cells by either expressing our MR1 construct directly in these cell lines or by adding their lysate into the media of transfected producing cells; both techniques that we have used before. Identification of the ligands loaded into MR1 from these experiments will be determined by LC-MS/MS through our collaboration with the Hildebrand Lab at the Oklahoma University Health Sciences Center. Following this determination, we will capitalize on our structural biology expertise to determine how potential ligands bind the MR1 pocket. Finally, we will use MR1/TAA tetramers to stain MR1T clones from donor blood samples and investigate their transcriptional phenotype to shed light on the cognate T cell population that is reactive to these antigens.

项目摘要： 长期以 先天和适应性免疫菌株种群。超越了肽的经典MHC I和II 分子，在人类中存在多种其他与结构相关的蛋白质 各种T细胞子类型的抗原类型。其中是单态MHC相关蛋白1 （MR1）在历史上以微生物衍生的核黄素代谢物呈现给粘膜 - 相关的不变T（MAIT）细胞。扩大了这一初步观察结果，我们的小组和其他人表明 MR1的核心体比以前所欣赏的要多样化。 MR1能够呈现配体 除了最初描述的肋骨代谢物以外的各种尺寸和化学结构 携带完全重排的αβTCR的MR1限制的T细胞（MR1TS）。最近，有新兴报告 MR1T克隆在完整中显示了针对取消时间的MR1依赖性细胞毒性活性的 没有任何微生物感染或微生物配体。由于这些观察的前所未有的性质 以及基于MR1的免疫疗法的潜在影响，重要的是配体是由 在癌症引起的细胞功能下，MR1。考虑到我们自己的 研究和这些新颖的报道，我们建议MR1充当两者中细胞代谢组的传感器 微生物感染和癌症等内源性失调的背景，呈现了一系列代谢产物 抗原。由于我们既定的往绩记录，调查了MR1的生物化学及其现有协作 MR1的配体发现管道，Adams Lab是独特的中毒以研究抗原的身份 MR1在癌症的背景下呈现。我们的投资将重点放在血浆细胞癌上，多个 骨髓瘤，因为该癌症的细胞系已显示在细胞上具有绝对高的MR1表达 表面，表明较高的抗原负荷。我们建议我们已建立的微生物配体发现 可以调整MR1的管道来研究与肿瘤相关的抗原（TAA） 通过直接在这些细胞系中表达我们的MR1构建的骨髓瘤细胞表面，或者通过添加它们 裂解物进入翻译的生产细胞的培养基；我们以前使用过的两种技术。识别 这些实验中加载到MR1中的配体将由我们的协作确定LC-MS/MS 在俄克拉荷马大学健康科学中心的希尔德布兰德实验室。遵循此决定，我们 将利用我们的结构生物学专业知识，以确定潜在配体如何结合MR1袋。最后， 我们将使用MR1/TAA四聚体从供体血液样本中染色MR1T克隆，并调查其 转录表型阐明了对这些抗原反应的同源T细胞群。