Identification and characterization of molecules important in the immune system

免疫系统中重要分子的鉴定和表征

• 批准号: 7284140
• 负责人: Nami McCarty
• 金额: $ 10.8万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-06 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7284140
• 关键词: Adaptor Signaling Protein; Apoptotic; Autoimmune Diseases; Biological Process; Bypass; CD4 Positive T Lymphocytes; Cantor; Cell Survival; Cell surface; Cells; Cellularity; Computer Analysis; Dana-Farber Cancer Institute; Data; Development; Dominant-Negative Mutation; Experimental Autoimmune Encephalomyelitis; Experimental Designs; Gene Expression; Genes; Genetic; Genomics; Grant; Immune response; Immune system; Individual; K22 Award; Knockout Mice; Light; Lymphoid; Methodology; Methods; Molecular; Molecular Profiling; Mus; Pathway interactions; Patients; Peripheral; Phosphotransferases; Play; Population; Process; Proteins; RNA Interference; RXR; Recycling; Regulation; Research; Role; Self Tolerance; Signaling Molecule; Stem cells; Subfamily lentivirinae; T-Cell Development; T-Lymphocyte; Therapeutic; Thymocyte Development; Thymus Gland; Time; Transcript; Work; base; chemokine; cytokine; gene function; human RIPK1 protein; human disease; knock-down; novel; reconstitution; serial analysis of gene expression; small hairpin RNA; thymocyte

DESCRIPTION (provided by applicant): Understanding the genetic mechanisms that promote lineage commitment and eliminate autoreactive thymocytes in the thymus has direct relation to human disease. My research involves identifying new molecules important in the development and differentiation of the thymus, and characterizing their molecular basis in regulating immune responses to both self and non-self. Studying and understanding self-tolerance mechanisms may result in therapeutic treatments for patients with harmful autoimmune diseases. My experimental design involves the identification and characterization of novel molecules necessary for proper cell development and differentiation. I use genomics (Serial Analysis of Gene Expression) to identify specific gene products differentially expressed in distinct T cell populations. I then characterize the molecules with a novel RNAi method to generate gene-specific deficient mice. In brief, lentivirus containing gene specific shRNA is introduced into purified hematopoetic stem cells (HSC's). RAG2 -/- recipient mice are then reconstituted with HSC's to generate gene-specific knockdown mice. With these combined genomic and RNAi-based methodologies I identified and characterized MINK (Misshapen-NIK-related kinase), which is important for negative selection in the thymus. Further, I aim to extend the use of these approaches to characterize other molecules identified by SAGE. This research will contribute to identifying new molecules important for immune responses and self-tolerance. Moreover, the rapid characterization of their biological functions will help to understand and find the proper targets for treating autoimmune diseases.

描述（由申请人提供）：了解促进谱系定型和消除胸腺中自身反应性胸腺细胞的遗传机制与人类疾病有直接关系。我的研究涉及鉴定在胸腺发育和分化中重要的新分子，并表征它们在调节对自身和非自身的免疫反应中的分子基础。研究和了解自我耐受机制可能会为患有有害自身免疫性疾病的患者提供治疗方法。 我的实验设计涉及适当细胞发育和分化所需的新分子的识别和表征。我使用基因组学（基因表达的连续分析）来 识别在不同 T 细胞群中差异表达的特定基因产物。然后我用一种新的 RNAi 方法来表征这些分子，以产生基因特异性缺陷的小鼠。简而言之，将含有基因特异性shRNA的慢病毒引入纯化的造血干细胞(HSC)中。然后用 HSC 重组 RAG2 -/- 受体小鼠以产生基因特异性敲除小鼠。 通过这些组合的基因组和基于 RNAi 的方法，我识别并表征了 MINK （Misshapen-NIK 相关激酶），对于胸腺中的负选择很重要。此外，我的目标是扩展这些方法的用途，以表征 SAGE 识别的其他分子。 这项研究将有助于识别对免疫反应重要的新分子 自我宽容。此外，对其生物学功能的快速表征将有助于理解和找到治疗自身免疫性疾病的适当靶点。