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 这项K08计划的目标是提供教学和经验培训所需的新技术 实现我的长期职业目标，即开发一个探索小说的独立研究计划 长非编码RNA(LncRNA)和同种异体免疫的交集。拟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，鉴定lncRNA-蛋白质的技术 互动、赠款撰写、指导和实验室管理。这项为期5年的培训计划将指导 我的主要导师，帕万·雷迪博士，他是一位成就卓著的移植免疫学内科科学家 有成功指导初级教员的记录。我还将得到一个有专业知识的委员会的建议 生物信息学、RNA生物学、分子遗传学和造血干细胞移植免疫学。说教培训计划 涵盖生物信息学、拨款撰写、高级免疫学、导师和实验室管理。 异基因(Allo)HSCT是一种治疗高危血液病的根治方法。急性移植物抗宿主 疾病(GVHD)是allo-HSCT的一种由allo-T细胞驱动的主要并发症，针对这种并发症改进的治疗方法有 需要的。LncRNAs控制具有组织特异性的基因表达并微调免疫反应。要确定 关于allo-T细胞潜在的lncRNA调节因子，我们最近在有良好注解的临床上进行了RNA测序 HSCT样本。本工作鉴定了LINC00402是一种新的、保守的、富含T细胞的lncRNA 由allo-T细胞差异表达。在功能上，LINC00402促进了allo-T细胞的增殖。然而，它 目前尚不清楚LINC00402是否会在体内加重急性GVHD，以及相关的分子机制 对异基因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，这项提议 将验证LINC00402通过增强ERK-c-fos在体内调节急性GVHD的总体假设 CD41型(Th1)辅助细胞中的信号转导。这将在体外用互补的小鼠和人类进行测试。 和活体系统。目标1将确定LINC00402在T细胞反应中的细胞机制，并将 检验LINC00402促进Th1分化和Th1细胞因子产生的特异性假设 通过异基因T细胞。目的2将明确介导LINC00402‘S调节T细胞和 将检验LINC00402直接增强ERK-c-fos依赖的T细胞受体信号的假设。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 以及识别lncRNA结合伙伴的方法。目标3将阐明LINC00402在实验中的作用 急性胃肠道GVHD，并将探索LINC00402增强急性胃肠道疾病的假设 GVHD通过促进Th1细胞的积聚。这将用互补的异种和 同种异体小鼠模型。总之，这些研究将探索同种异体免疫的一个独特领域，并确定 LINC00402是改善allo-HSCT结局的靶点。