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

基本信息

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
项目类别：
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

项目摘要

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：主要组织相容性复合物 (MHC) 分子长期以来一直被认为是之间的桥梁超越肽呈递的经典 MHC I 类和 II 类细胞群。分子中，人类体内还存在多种其他结构相关的蛋白质，已被发现呈现多种 T 细胞亚型的不同抗原类型就是其中之一。（MR1）历史上因将微生物来源的核黄素代谢物呈递至粘膜-而闻名我们的团队和其他人在这一初步观察的基础上扩展了相关不变 T (MAIT) 细胞。 MR1 的配体组比之前认为的 MR1 能够呈递的配体更加多样化。除了最初描述的核糖代谢物之外，还具有各种大小和化学结构携带完全重排的αβTCR的MR1限制性T细胞（MR1T）最近不断有报道。的 MR1T 克隆在完整的研究中显示出针对癌组织的 MR1 依赖性细胞毒活性由于这些观察结果是前所未有的，因此不存在任何微生物感染或微生物配体。以及基于 MR1 的免疫疗法的潜在影响，重要的是，由考虑到我们自己的情况，MR1 在癌症引起的细胞功能障碍中被识别。研究和这些新颖的报告中，我们提出 MR1 作为细胞代谢组的传感器微生物感染和癌症等内源性功能障碍的背景，呈现出多种代谢物由于我们已建立的研究 MR1 生物化学的记录以及我们现有的合作。 MR1 的配体发现管道，Adams 实验室专门准备研究抗原的身份 MR1 是在癌症背景下出现的，我们的研究将集中于浆细胞癌、多发性癌症。骨髓瘤，因为这种癌症的细胞系已被证明在细胞中具有异常高的 MR1 表达表面，表明可能存在更高的抗原负载。 MR1 的管道可用于研究呈递在 MR1 上的肿瘤相关抗原 (TAA) 通过直接在这些细胞系中表达我们的 MR1 构建体或通过添加它们的裂解物进入转染的生产细胞的培养基中；这两种技术都是我们之前使用过的。通过我们的合作，将通过 LC-MS/MS 确定从这些实验加载到 MR1 中的配体根据这一决定，我们与俄克拉荷马大学健康科学中心的希尔德布兰德实验室合作。将利用我们的结构生物学专业知识来确定潜在的配体如何结合 MR1 口袋。我们将使用 MR1/TAA 四聚体对供体血液样本中的 MR1T 克隆进行染色，并研究它们的转录表型以揭示对这些抗原有反应的同源 T 细胞群。