Signaling pathways that interface with Klf2 to regulate B cell migration

与 Klf2 相互作用调节 B 细胞迁移的信号通路

基本信息

批准号：
8636873
负责人：
Eric Sebzda
金额：
$ 23.55万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-15 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8636873
关键词：
Affect American Animal Model Applications Grants Arthritis Autoimmune Diseases Autoimmunity B-Lymphocytes Binding Biochemical Genetics Blood Blood Circulation Body cavities Cell Lineage Cells Clinical Trials Complex DNA Microarray Chip Data Diabetes Mellitus Disease Disease Progression Drug Design Ensure Event Excision Gene Expression Gene Proteins Gene Targeting Genes Genetic Genetic Programming Genetic Transcription Goals High-Throughput DNA Sequencing Home environment Homeostasis Homing Human Immune response Individual Insulin-Dependent Diabetes Mellitus Investigation Knowledge Link Location Lupus Lymphocyte Lymphoid Mass Spectrum Analysis Mature B-Lymphocyte Mediating Microarray Analysis Microbe Molecular Molecular Profiling Molecular Target Mus Nuclear Nucleic Acid Regulatory Sequences Organ Pattern Physiological Population Promoter Regions Protein Binding Proteins Proteomics Receptor Gene Regulation Research Project Grants Research Proposals Rheumatoid Arthritis Self Tolerance Shapes Signal Pathway Signal Transduction Spleen Surface System Systemic Lupus Erythematosus Tamoxifen Techniques Technology Testing Viral autoreactive B cell base body cavity catalyst cell motility chromatin immunoprecipitation design gene discovery insight lupus-like migration mouse model novel pathogen prevent protein complex public health relevance receptor receptor expression research study small hairpin RNA therapeutic target trafficking transcription factor

项目摘要

DESCRIPTION (provided by applicant): B cell-depletion therapies in humans as well as accumulating evidence from animal models indicate that B lymphocytes help initiate several autoimmune disorders including arthritis, lupus, and type 1 diabetes. Recent clinical trials have shown that restricting lymphocyte migration can ameliorate autoimmune diseases, raising the possibility that B cell circulation can be targeted as a means of treating auto reactivity. A major barrier towards achieving this goal is the dearth of mechanistic knowledge concerning the regulation of B cell homing receptors. This R21 project is predicated on the applicant's discovery that Kruppel-like factor 2 (Klf2) is a key transcription factor that controls mature B cel migration patterns. Moreover, Klf2 excision within the B cell compartment results in a lupus-like form of autoimmunity that further supports the link between altered B cell migration and disease progression. The premise of the current grant proposal is that Klf2 controls several important homing receptors and that by defining the genes and proteins that interface with Klf2, the molecular mechanisms that orchestrate B cell trafficking will be revealed. To achieve this goal, Aim 1 studies will initially use quantitative mass-spectrometry (MS) techniques to identify interactive proteins that facilitate Klf2 function. Parallel MS studies will define the protein complexes bound to the promoter regions of Klf2-regulated homing receptors. Together, these experiments will reveal a subset of Klf2-interactive proteins that directly shape B cell migration in a linage-specific manner. In Aim 2 studies, genes under Klf2 control that regulate B cell trafficking will be captured using a combination of DNA microarrays and chromatin immunoprecipitation followed by high-throughput DNA sequencing (ChIP-Seq) approaches. These results will define the genetic footprint of Klf2 and thus provide a cache of molecular targets involved in B cell homeostasis and self- tolerance. Access to primary B cell lineages that lack Klf2 provide the ideal control for the proposed biochemical and genetic experiments. Overall results emerging from the proposed project will provide the first insight into how mature B cell circulation patterns are maintained. In turn, these discoveries may promote the rationale design of new therapies to control B cell trafficking as a means of ameliorating disease.

描述（由申请人提供）：B细胞消耗疗法以及从动物模型中积累的证据表明，B淋巴细胞有助于引发多种自身免疫性疾病，包括关节炎，狼疮和1型糖尿病。最近的临床试验表明，限制淋巴细胞迁移可以改善自身免疫性疾病，从而提高了B细胞循环作为治疗自身反应性的一种手段的可能性。专业实现这一目标的障碍是缺乏有关调节B细胞归纳受体的机械知识。这个R21项目是基于申请人发现的，即Kruppel样因子2（KLF2）是控制成熟的B Cel迁移模式的关键转录因子。此外，B细胞室内的KLF2切除会导致类似狼疮的自身免疫力形式，进一步支持B细胞迁移与疾病进展的改变之间的联系。当前赠款提案的前提是KLF2控制了几种重要的归巢受体，并且通过定义与KLF2接口的基因和蛋白质，将揭示精心的B细胞运输的分子机制。为了实现这一目标，AIM 1研究最初将使用定量质谱法（MS）技术来识别促进KLF2功能的交互式蛋白质。平行的MS研究将定义与KLF2调节的归因受体的启动子区域结合的蛋白质复合物。总之，这些实验将揭示一部分KLF2相互作用蛋白，这些蛋白直接以外线特异性方式塑造B细胞迁移。在AIM 2研究中，将使用DNA微阵列和染色质免疫沉淀的组合，然后进行高通量DNA测序（CHIP-SEQ）方法来捕获调节B细胞运输的基因。这些结果将定义KLF2的遗传足迹，从而提供了与B细胞稳态和自耐受性有关的分子靶标的缓存。进入缺乏KLF2的原代B细胞谱系为提出的生化和遗传实验提供了理想的控制。拟议项目中出现的总体结果将提供有关如何保持成熟B细胞循环模式的首次见解。反过来，这些发现可能会促进新疗法的基本原理设计，以控制B细胞运输作为改善疾病的一种手段。