Single Cell Analysis and Immunogenetics

单细胞分析和免疫遗传学

• 批准号: 10493796
• 负责人: Kenneth James Livak
• 金额: $ 30.82万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10493796
• 关键词: Address; Algorithms; Antigens; Applications Grants; Binding; Bronchiolitis Obliterans; Cell Line; Cell physiology; Cells; Characteristics; Chronic; Clinical; Cloning; DNA; DNA analysis; DNA sequencing; Data; Data Analyses; Databases; Disease; Disease model; Genetic Transcription; Goals; Hematopoietic Stem Cell Transplantation; Immune; Immune response; Immunogenetics; Immunogenomics; Immunologics; Individual; Institutes; Laboratories; Lung; Metabolic; Minor Histocompatibility Antigens; Mus; Outcome; Pathogenesis; Pathway interactions; Patients; Peptides; Population; Principal Investigator; Resistance; Sampling; Single Nucleotide Polymorphism; Specimen; Syndrome; T cell clonality; T cell response; T-Cell Immunologic Specificity; T-Cell Receptor; T-Lymphocyte; T-cell receptor repertoire; Testing; Therapeutic; Tissues; Variant; alpha-beta T-Cell Receptor; beta Chain Antigen T Cell Receptor; chronic graft versus host disease; computerized tools; exome sequencing; graft vs host disease; immunogenicity; insight; mouse model; new technology; response; single cell analysis; single cell sequencing; tool; transcriptome sequencing; treatment response; tumor-immune system interactions

Project Summary The projects proposed for this grant application seek to characterize response and resistance to treatments for chronic graft versus host disease (cGVHD) following hematopoietic stem cell transplantation (HCT). Core 1 will support the single cell- and immunogenomics-related goals of these projects by focusing on applying the latest experimental and computational tools to these studies. Bulk and single-cell transcriptome sequencing of immune and lung cell populations (Aim 1) will be used to identify relevant pathways related to response to treatment of cGVHD, to identify transcriptional aspects of the metabolic signature elucidated in mouse lung cGVHD models, and to characterize the lung pathogenesis and immune response related to Bronchiolitis Obliterans Syndrome (BOS) following HCT. Response of T cells to cGVHD treatments and to BOS will be further characterized by TCR repertoire analysis using targeted bulk and single-cell sequencing to assess T cell clonality (Aim 2). Core 1 will analyze whole exome sequencing data from donor and recipient DNA to identify polymorphic differences between donor and recipient, and use tissue expression databases to determine which of these variants are expressed in lung. Recently developed sophisticated algorithms will be implemented to use this information to predict personal HLA-binding peptides that comprise minor histocompatibility antigens (Aim 3). The paired alpha/beta TCR chain single-cell sequence information will be used to reconstruct cell lines expressing individual enriched TCRs (Aim 4) in order to functionally determine exactly which TCR interacts with which antigen. This analysis will directly assess if mHAg-directed T cell responses contribute to clinical responses to therapy.

项目概要 本次拨款申请提出的项目旨在描述对治疗的反应和抵抗 造血干细胞移植（HCT）后的慢性移植物抗宿主病（cGVHD）。核心1将 通过专注于应用最新的技术来支持这些项目的单细胞和免疫基因组学相关目标 这些研究的实验和计算工具。免疫的批量和单细胞转录组测序 和肺细胞群（目标 1）将用于确定与治疗反应相关的相关途径 cGVHD，鉴定小鼠肺 cGVHD 模型中阐明的代谢特征的转录方面， 并表征与闭塞性细支气管炎综合征相关的肺部发病机制和免疫反应 (BOS) HCT 后。 T 细胞对 cGVHD 治疗和 BOS 的反应将进一步表征为 使用靶向批量和单细胞测序进行 TCR 谱分析以评估 T 细胞克隆性（目标 2）。核 1 将分析来自供体和受体 DNA 的全外显子组测序数据，以识别多态性差异 供体和受体之间的差异，并使用组织表达数据库来确定这些变异中的哪些是 表达于肺。最近开发的复杂算法将被实施，以使用这些信息 预测包含次要组织相容性抗原的个人 HLA 结合肽（目标 3）。配对的 α/β TCR 链单细胞序列信息将用于重建表达个体的细胞系 富集 TCR（目标 4），以便在功能上准确确定哪个 TCR 与哪个抗原相互作用。这 分析将直接评估 mHAg 导向的 T 细胞反应是否有助于临床治疗反应。