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

基本信息

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
项目类别：
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

项目摘要

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的长期职业目标，即开发探索小说的独立研究计划长非编码 RNA (lncRNA) 和同种免疫的交叉点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 蛋白的技术该 5 年培训计划将指导互动、资助写作、指导和实验室管理。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，我还将得到一个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。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 LncRNA 通过组织特异性控制基因表达并微调免疫反应。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 测序这项工作鉴定出 LINC00402 是一种新型、保守、富含 T 细胞的 lncRNA。在功能上，LINC00402 促进了 allo-T 细胞增殖。尚不清楚 LINC00402 是否会使体内急性 GVHD 恶化以及是什么分子机制造成的鉴于其对异基因 T 细胞功能的影响，该提案基于这些先前数据和其他新观察结果。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体外进行测试。目标 1 将确定 LINC00402 在 T 细胞反应中的细胞机制。测试 LINC00402 增强 Th1 分化和 Th1 细胞因子产生的具体假设目标 2 将定义介导 LINC00402 对 T 细胞的调节的分子机制。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。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总而言之，这些研究将探索同种异体免疫的独特领域并确定是否存在。 LINC00402 是改善异基因造血干细胞移植结果的靶标。