Non-coding RNAs in B lymphocyte development and function

B 淋巴细胞发育和功能中的非编码 RNA

• 批准号: 8686366
• 负责人: Sergei Borisovich Koralov
• 金额: $ 16.95万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-18 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8686366
• 关键词: Ablation; Address; Alleles; Animal Model; Animals; Antibodies; Antibody Repertoire; Antigen Receptors; Apoptosis; B cell differentiation; B-Cell Development; B-Lymphocytes; Binding; Bone Marrow; Cell Line; Cell Separation; Cell Survival; Cell surface; Cells; Chromatin; Complex; DNA Sequence Rearrangement; Data; Development; Elements; Event; Floods; Functional RNA; Future; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Recombination; Genetic Transcription; Goals; Immune; Immune system; Immunoglobulin Class Switching; Immunoglobulin Genes; Immunoglobulin Somatic Hypermutation; Immunoglobulin Switch Recombination; Immunoglobulins; Individual; Laboratories; Light; Lymphocyte; Lymphomagenesis; Maintenance; Mature B-Lymphocyte; Medical; Membrane; MicroRNAs; Microprocessor; Mus; Nature; Nuclear; Nuclear RNA; Organ; Pathway interactions; Physiological; Plasma Cells; Play; Population; Postdoctoral Fellow; Process; Proteins; RNA; RNA Interference; Reaction; Receptor Gene; Receptors, Antigen, B-Cell; Recovery; Regulation; Research; Role; Sampling; Science; Signal Pathway; Signal Transduction; Staging; Structure of germinal center of lymph node; Students; System; Systems Development; Time; Transcriptional Regulation; Transgenes; Transgenic Organisms; V(D)J Recombination; Work; animal facility; base; cell type; chromatin modification; human DICER1 protein; interest; kappa-Chain Immunoglobulins; mouse model; novel; plasma cell differentiation; progenitor; programs; response; tool

Regulation of recombination and further diversification of immunoglobulin genes by class switch recombination and somatic hypermutation is tightly controlled in a cell type, stage specific and allele specific manner. The precise mechanisms that regulate chromatin accessibility of antigen receptor genes during these events remain elusive. Our goal is to evaluate the role of RNAi in B cell development and function. We have previously demonstrated that miRNAs are essential for B cell survival. Specifically, ablation of miRNAs in B cells upon deletion of Dicer, Drosha or DGCR8 blocked the pro-B to pre-B cell transition due to an increase in apoptosis. By rescuing B cell development using a pro-survival Bcl2 transgene, we are able to investigate the role of short non-coding RNAs in V(D)J recombination as well as in later stages of B cell differentiation. Preliminary data from these studies reveals that ablation of Dicer impacts primary immune repertoire and that this does not happen when Drosha is deleted in a similar manner, suggesting a role for Dicer dependent non-coding RNAs in direct regulation of immunoglobulin loci. We will extend our studies to investigate the nature and role of these non-coding RNAs and examine the contribution of RNAi to immunoglobulin locus accessibility during the germinal center reaction. The control of the immunoglobulin loci by Dicer dependent, microprocessor complex independent RNAi mechanism represents a novel mechanism in mammalian locus regulation. Our goal is to understand the regulation of chromatin accessibility in this context. In addition, the ability to rescue B cell development in the absence of RNAi gives us an opportunity to examine the role of non-coding RNAs in terminal B cell differentiation and the role of miRNA in plasma cell generation – as the regulation of the terminal B cell differentiation program remains poorly understood. Our work suffered a significant hit as a result of superstorm Sandy as all the animal models used for these studies were lost with the flooding of the animal facility. In addition, our laboratory was forced to relocate following the storm due to significant damage to our building. This application outlines our efforts to recover the complex animal models necessary for our research and to reproduce the exciting preliminary data that we need to submit a competitive RO1 application within the next one to two years. Specifically, we will generate inducible pluripotent cells (iPS) from the sorted B cells of stored bone marrow samples of our original mice to generate the necessary mouse models for this research. The approach of using B cell derived iPS cells with conditional Dicer and DGCR8 alleles will recapitulate our ability to investigate the role of non-coding RNAs in the later stages of B cell development and give us an opportunity to study the role of non-coding RNAs in somatic hypermutation and class switch recombination in the context of several uniquely rearranged immunoglobulin loci.

通过类转换调节重组和进一步多样化免疫球蛋白基因 重组和体细胞超名在细胞类型，特定于阶段和等位基因中受到严格控制 特定的方式。 在事件期间仍然难以捉摸。 功能。 删除DICER，DROSHA或DGCR8后，miRNA在B细胞中的消融，阻止了pro- 由于凋亡的增加而导致的过渡。 转基因，我们要吓to（d）J重组中的短非编码RNA的作用 在B细胞分化的后期阶段。 影响原发性免疫免疫曲目，并且当Drosha被删除时不会发生 方式，表明DICER decent decentent depent非编码RNA在直接调节免疫球蛋白中的作用 基因座。 RNAi对生殖中心现实中免疫球蛋白基因座的贡献 通过DICER依赖，微处理器复合物独立RNAi控制免疫球蛋白基因座 抑制哺乳动物基因座调节的新机制。 在这种情况下，调节染色质的可及性。 在没有RNAi的情况下，我们有机会检查非编码RNA在末端B细胞中的作用 miRNA在浆细胞产生中的区分和作用 - 作为末端B细胞的调节 区分计划仍然很糟糕。 UREK像所有使用的动物模型一样，遭受了Asuperstorm Sandy的重大打击 随着动物设施的洪水泛滥，我们的实验室被迫搬迁。 由于对我们的建筑物的重大损害，风暴概述了我们 恢复我们研究所需的复杂动物模型，并重现令人兴奋的 我们需要在接下来的一到两次中提交竞争性RO1申请的初步数据 多年。 我们原始小鼠的骨髓样品为此而生成了必不可少的慕斯 使用有条件的DICR8等位基因Willeles Willeles使用B细胞衍生的IPS细胞的方法将使我们的 能够研究非编码RNA在B细胞发育后期的作用的能力，并给我们一个 研究非编码RNA在躯体超成名和类开关重组中的作用的机会 在几个独特重新排列的免疫球蛋白基因座的背景下。