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 K08 提案的目标是提供新技术所需的教学和体验培训 实现我开发探索小说的独立研究计划的长期职业目标 长非编码 RNA (lncRNA) 和同种免疫的交叉。拟议的培训将使我能够 发展造血干细胞移植（HSCT）动物模型、T细胞功能方面的专业知识 测定、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、资助写作、指导和实验室管理。本次5年培训计划将定向 由我的主要导师 Pavan Reddy 博士撰写，他是一位卓有成就的移植免疫学医师科学家 拥有成功指导初级教师的记录。我还将得到具有专业知识的委员会的建议 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、资助写作、高级免疫学、指导和实验室管理。 同种异体 (allo) HSCT 是一种治疗高危血液系统疾病的方法。急性移植物抗宿主 疾病（GVHD）是异基因 T 细胞驱动的异基因 HSCT 的主要并发症，对此的治疗方法正在改进 需要。 LncRNA 通过组织特异性控制基因表达并微调免疫反应。识别 同种异体 T 细胞的潜在 lncRNA 调节因子，我们最近对注释良好的临床进行了 RNA 测序 HSCT 样本。这项工作将 LINC00402 确定为一种新型、保守、富含 T 细胞的 lncRNA， 同种异体 T 细胞差异表达。从功能上讲，LINC00402 促进同种异体 T 细胞增殖。然而，它 尚不清楚 LINC00402 是否会加剧体内急性 GVHD 以及是什么分子机制造成的 因其对同种异体 T 细胞功能的影响。根据这些先前的数据和其他新的观察结果，该提案 将检验 LINC00402 通过增强 ERK-c-FOS 调节体内急性 GVHD 的总体假设 CD4 1 型 (Th1) 辅助细胞中的信号传导。这将使用互补的小鼠和人类进行体外测试 和体内系统。目标 1 将确定 LINC00402 在 T 细胞反应中的细胞机制，并将 测试 LINC00402 增强 Th1 分化和 Th1 细胞因子产生的具体假设 由同种异体 T 细胞。目标 2 将定义介导 LINC00402 对 T 细胞和 将检验 LINC00402 直接增强 ERK-c-FOS 依赖性 T 细胞受体信号传导的假设。它 还将作为学习下一代测序数据集生物信息学分析的培训平台 以及识别 lncRNA 结合伴侣的方法。目标 3 将阐明 LINC00402 在实验中的作用 急性胃肠道GVHD，并将探讨LINC00402增强急性胃肠道GVHD的假设 GVHD 通过促进 Th1 细胞的积累。这将使用互补的异种和 同种异体小鼠模型。总而言之，这些研究将探索同种免疫的独特领域，并确定是否 LINC00402 是改善异基因造血干细胞移植结果的靶标。