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: 10734891
• 负责人: Daniel C. Peltier
• 金额: $ 13.93万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10734891
• 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 differentiation; 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; 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 该K08提案的目标是提供所需的新技术的教学和经验丰富的培训 实现我的长期职业目标，即制定一个独立研究计划，探索小说 长的非编码RNA（LNCRNA）和同种免疫的交点。拟议的培训将使我能够 在造血干细胞移植（HSCT），T细胞功能的动物模型中发展专业知识 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，赠款写作，指导和实验室管理。这个为期5年的培训计划将被指导 由我的主要导师帕文·雷迪（Pavan Reddy）博士，他是一位出色的移植免疫学医生 - 科学家 具有成功的初级教师的记录。我还将由具有专业知识的委员会告知 在生物信息学，RNA生物学，分子遗传学和HSCT免疫学中。教学培训计划 涵盖生物信息学，赠款写作，先进的免疫学，指导和实验室管理。 同种异体（Allo）HSCT是一种治疗高危血液学疾病的治疗方法。急性移植物与主持人 疾病（GVHD）是Allo-T细胞驱动的Allo-HSCT的主要并发症，为此改善的治疗方法是 需要。 LNCRNAS以组织特异性和微调免疫回报控制基因表达。识别 我们最近对Allo-T细胞的潜在lncRNA调节剂进行 HSCT样品。这项工作将linc00402确定为一种小说，配置，富含T细胞的lncRNA 由异晶细胞差异表达。从功能上讲，Linc00402促进了异晶细胞增殖。但是，它 尚不清楚Linc00402是否加剧了体内急性GVHD和哪些分子机制是负责的 它对Allo-T细胞功能的影响。基于这些先前的数据和其他新观察结果，该建议 将测试Linc00402通过增强ERK-C-FOS在体内调节急性GVHD的总体假设 CD4型1（TH1）辅助细胞中的信号传导。这将在体外使用完整的鼠和人类进行测试 和体内系统。 AIM 1将确定T细胞反应中Linc00402的细胞机制，并将 检验Linc00402增强Th1分化和产生Th1细胞因子的特定假设 由Allo-T细胞。 AIM 2将定义介导Linc00402调节T细胞和的分子机制 将测试Linc00402直接增强ERK-C-FOS依赖性T细胞接收器信号传导的假设。它 还将作为学习下一代测序数据集的生物信息学分析的培训平台 和识别lncRNA结合伴侣的方法。 AIM 3将阐明Linc00402在实验中的作用 急性胃肠道GVHD，并将探讨Linc00402增强急性胃肠道的假设 GVHD通过促进Th1细胞的积累。这将使用完整的异构化和 同种异体鼠模型。总的来说，这些研究将探索同种免疫的独特领域，并确定是否是否 Linc00402是改善异含量HSCT结果的目标。