Long noncoding RNA-mediated regulation of T-cell alloimmunity

长非编码RNA介导的T细胞同种免疫调节

• 批准号: 10368203
• 负责人: Daniel C. Peltier
• 金额: $ 13.89万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 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 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10368203
• 关键词: Acute; Acute Graft Versus Host Disease; Allogenic; Animal Model; Area; Binding; Bioinformatics; Biological Assay; Biology; Cell Nucleus; Cell physiology; Cells; Clinical; Complication; Cytoplasm; Cytotoxic T-Lymphocytes; Data; Data Set; Development Plans; Disease model; Enhancers; FOS gene; Faculty; Gene Expression; Genetic Transcription; Goals; Grant; Hematological Disease; Hematopoietic Stem Cell Transplantation; Human; Immune response; Immunology; In Vitro; Interleukin-2; Laboratories; Learning; Measures; Mediating; Mentors; Mentorship; Methods; Molecular; Molecular Genetics; Mus; Outcome; Patients; Physicians; Production; Proteins; RNA; RNA Binding; RNA-Protein Interaction; Receptor Signaling; Regulatory T-Lymphocyte; Research; Research Personnel; Role; Sampling; Scientist; Signal Transduction; Specificity; System; T cell regulation; T cell response; T-Cell Activation; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Th1 Cells; Tissues; Training; Training Programs; Transplantation Immunology; Untranslated RNA; Work; Writing; base; career; career development; cell killing; cohort; curative treatments; cytokine; cytotoxic; cytotoxic CD8 T cells; differential expression; gastrointestinal; graft vs host disease; high risk; improved; in vivo; isoimmunity; mouse model; next generation sequencing; novel; programs; response; transcriptome sequencing

Project Abstract The goal of this K08 proposal is to provide didactic and experiential training in new techniques needed to achieve my long-term career goal of developing an independent research program exploring the novel intersection of long noncoding RNAs (lncRNA) and alloimmunity. The proposed training will allow me to develop expertise in animal models of hematopoietic stem cell transplantation (HSCT), T cell functional assays, bioinformatics analysis of next-generation sequencing data sets, techniques to identify lncRNA-protein interactions, grant writing, mentorship, and laboratory management. This 5-year training plan will be directed by my primary mentor, Dr. Pavan Reddy, who is an accomplished transplant immunology physician-scientist with a track record of successfully mentoring junior faculty. I will also be advised by a committee with expertise in bioinformatics, RNA biology, molecular genetics, and HSCT immunology. The didactic training program covers bioinformatics, grant writing, advanced immunology, mentorship, and laboratory management. Allogeneic (allo) HSCT is a curative treatment for high-risk hematologic disorders. Acute graft-versus-host disease (GVHD) is an allo-T cell-driven major complication of allo-HSCT, for which improved treatments are needed. LncRNAs control gene expression with tissue specificity and fine-tune immune responses. To identify potential lncRNA regulators of allo-T cells, we recently performed RNA-sequencing on well-annotated clinical HSCT samples. This work identified LINC00402 as a novel, conserved, T cell-enriched lncRNA that was differentially expressed by allo-T cells. Functionally, LINC00402 promoted allo-T cell proliferation. However, it is unknown if LINC00402 exacerbates acute GVHD in vivo and what molecular mechanisms are responsible for its effects on allo-T cell function. Based on these prior data and additional new observations, this proposal will test the overall hypothesis that LINC00402 regulates acute GVHD in vivo by enhancing ERK-c-FOS signaling in CD4 type 1 (Th1) helper cells. This will be tested using complementary murine and human in vitro and in vivo systems. Aim 1 will determine the cellular mechanisms of LINC00402 in T cell responses and will test the specific hypothesis that LINC00402 augments Th1 differentiation and the production of Th1 cytokines by allo-T cells. Aim 2 will define the molecular mechanisms mediating LINC00402’s regulation of T cells and will test the hypothesis that LINC00402 directly enhances ERK-c-FOS-dependent T cell receptor signaling. It will also serve as a training platform to learn bioinformatics analysis of next-generation sequencing datasets and methods to identify lncRNA binding partners. Aim 3 will elucidate the role of LINC00402 in experimental acute gastrointestinal GVHD, and will explore the hypothesis that LINC00402 enhances acute gastrointestinal GVHD by promoting accumulation of Th1 cells. This will be tested using complementary xenogeneic and allogeneic murine models. Altogether, these studies will explore a unique area of alloimmunity and determine if LINC00402 is a target for improving allo-HSCT outcomes.

项目摘要 本K 08提案的目标是提供所需新技术的教学和体验培训， 实现我的长期职业目标，开发一个独立的研究计划，探索小说 长链非编码RNA（lncRNA）与同种异体免疫的交叉。建议的培训将使我能够 发展造血干细胞移植（HSCT）动物模型的专业知识，T细胞功能 分析，下一代测序数据集的生物信息学分析，识别lncRNA蛋白的技术 互动，赠款写作，指导和实验室管理。这一5年培训计划将针对 我的主要导师Pavan Reddy博士是一位有成就的移植免疫学医生兼科学家 有着成功指导初级教员的记录我还将听取一个专家委员会的意见 生物信息学、RNA生物学、分子遗传学和HSCT免疫学。教学培训计划 涵盖生物信息学、资助撰写、高级免疫学、导师制和实验室管理。 同种异体（allo）HSCT是一种治疗高危血液系统疾病的有效方法。Acute graft-versus-host 移植物抗宿主病（GVHD）是异基因造血干细胞移植的一种主要并发症， needed. LncRNA以组织特异性控制基因表达并微调免疫应答。以识别 潜在的lncRNA调节剂的allo-T细胞，我们最近进行了RNA测序， HSCT样本。这项工作将LINC 00402鉴定为一种新的、保守的、富含T细胞的lncRNA， 异基因T细胞的差异表达。在功能上，LINC 00402促进同种异体T细胞增殖。但 LINC 00402是否在体内加重急性GVHD以及引起的分子机制尚不清楚 对同种异体T细胞功能的影响。根据这些先前的数据和额外的新观察， 将检验LINC 00402通过增强ERK-c-FOS在体内调节急性GVHD的总体假设 CD 4 1型（Th 1）辅助细胞中的信号传导。这将在体外使用互补的鼠和人进行测试 和体内系统。目的1将确定LINC 00402在T细胞应答中的细胞机制， 检验LINC 00402增强Th 1分化和Th 1细胞因子产生的特定假设 通过异基因T细胞。目的2将明确LINC 00402调节T细胞的分子机制， 将检验LINC 00402直接增强ERK-c-FOS依赖性T细胞受体信号传导的假设。它 还将作为一个培训平台，学习下一代测序数据集的生物信息学分析 和鉴定lncRNA结合配偶体的方法。目的3将阐明LINC 00402在实验中的作用 急性胃肠道GVHD，并将探索LINC 00402增强急性胃肠道GVHD的假设 GVHD通过促进Th 1细胞的积累。这将使用互补异种和 同种异体鼠模型。总之，这些研究将探索同种免疫的独特领域，并确定是否 LINC 00402是改善allo-HSCT结局的靶点。