The epigenetic reader BRWD1 in peripheral adaptive immunity

外周适应性免疫中的表观遗传阅读器 BRWD1

• 批准号: 10541126
• 负责人: Marcus Ramsay Clark
• 金额: $ 48.78万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-10 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541126
• 关键词: 3-Dimensional; ATAC-seq; Affect; Affinity; Alleles; Antibodies; Antigens; Applications Grants; Attenuated; Autoimmunity; B-Cell Development; B-Lymphocytes; Binding; Binding Sites; Bone Marrow; Cells; ChIP-seq; Characteristics; Chimera organism; Chromatin; Chromatin Structure; DNA; Darkness; Data; Defect; Development; Enhancers; Ephrin-A5; Epigenetic Process; Failure; Flow Cytometry; Follicular Dendritic Cells; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Heterogeneity; Hi-C; Histologic; Human; Humoral Immunities; IRF4 gene; IgK; Immunization; Immunize; Immunoglobulin Class Switching; In Situ; Infection; Knowledge; Light; Lymphopoiesis; Malignant Neoplasms; Molecular; Mus; Mutation; Nucleosomes; Patients; Pattern; Peripheral; Phenotype; Population; Positioning Attribute; Reader; Regulation; Reporting; Repression; Role; Shapes; Sorting; Structure; Structure of germinal center of lymph node; T-Lymphocyte; Testing; Transcriptional Regulation; WD Repeat; Work; adaptive immunity; design; differential expression; fighting; genome-wide; hypogammaglobulinemia; lymph nodes; molecular modeling; novel; novel therapeutics; programs; quantitative imaging; recruit; response; single-cell RNA sequencing; three dimensional structure; transcription factor; vaccine efficacy; 3-Dimensional; ATAC-seq; Affect; Affinity; Alleles; Antibodies; Antigens; Applications Grants; Attenuated; Autoimmunity; B-Cell Development; B-Lymphocytes; Binding; Binding Sites; Bone Marrow; Cells; ChIP-seq; Characteristics; Chimera organism; Chromatin; Chromatin Structure; DNA; Darkness; Data; Defect; Development; Enhancers; Ephrin-A5; Epigenetic Process; Failure; Flow Cytometry; Follicular Dendritic Cells; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Heterogeneity; Hi-C; Histologic; Human; Humoral Immunities; IRF4 gene; IgK; Immunization; Immunize; Immunoglobulin Class Switching; In Situ; Infection; Knowledge; Light; Lymphopoiesis; Malignant Neoplasms; Molecular; Mus; Mutation; Nucleosomes; Patients; Pattern; Peripheral; Phenotype; Population; Positioning Attribute; Reader; Regulation; Reporting; Repression; Role; Shapes; Sorting; Structure; Structure of germinal center of lymph node; T-Lymphocyte; Testing; Transcriptional Regulation; WD Repeat; Work; adaptive immunity; design; differential expression; fighting; genome-wide; hypogammaglobulinemia; lymph nodes; molecular modeling; novel; novel therapeutics; programs; quantitative imaging; recruit; response; single-cell RNA sequencing; three dimensional structure; transcription factor; vaccine efficacy

PROJECT SUMMARY Recently, we demonstrated that the lineage-restricted BROMO and WD40 domain containing epigenetic reader BRWD1 opens Igk and induces an epigenetic landscape that enables assembly of RAG proteins at Jk. We now report that BRWD1 has a much broader role in late B cell development. BRWD1 reshapes chromatin landscapes by closing enhancers of genes expressed early in B cell development and opening those of genes expressed in late stages. BRWD1 did not affect the expression of critical transcription factors (TFs). Rather, as demonstrated for IRF4, BRWD1 determines the sites they bind across the genome. BRWD1 differentially regulated over 7000 genes, repressing proliferative and inducing differentiation programs. Notably, BRWD1 coordinately repressed Myc and Myc target genes. Furthermore, in humans, BRWD1 mutations are associated with hypogammaglobulinemia. These data indicate that, in both mice and humans, BRWD1 is a master orchestrator of enhancer accessibility that cooperates with TF networks to drive late B cell development. BRWD1 is first expressed in small pre-B cells. However, Brwd1 expression is higher in naïve follicular (FO) and in germinal center (GC) dark zone (DZ) cells. Analysis of genes differentially expressed in GC DZ and light zone (LZ) cells, compared to those differentially regulated by BRWD1 in small pre-B cells, predicts that BRWD1 represses the DZ program and induces the LZ program. Furthermore, differential genome-wide binding of BRWD1 in DZ and LZ cells suggests different functions in each GC subset. Immunizing Brwd1fl/fl x Aicda (AID)- Cre mice revealed that BRWD1 was required for normal antibody isotype switching and affinity maturation. Histologically, GCs were greatly reduced in both size, and number, and lacked follicular dendritic cells. Therefore, we hypothesize that BRWD1 regulates epigenetic states and enhancer landscapes critical for coordination of DZ and LZ transcriptional programs and GC function. We will test this hypothesis in the following Specific Aims: Aim 1. Determine the transcriptional regulation of normal germinal center responses. Aim 2. Determine the role of BRWD1 in adaptive immunity. Aim 3. Determine the role of BRWD1 in initiating GC responses and in maintaining FO B cell identity. !

项目摘要 最近，我们证明了包含表观遗传读取器的谱系限制性Bromo和WD40领域 BRWD1打开IGK并诱导表观遗传景观，该景观使JK可以组装RAG蛋白。我们现在 报道BRWD1在B细胞晚期发育中起着更广泛的作用。 BRWD1重塑染色质景观 通过结束在B细胞发育早期表达的基因的增强子，并打开了在 晚期。 BRWD1不影响关键转录因子（TFS）的表达。相反，如证明 对于IRF4，BRWD1确定它们在基因组中结合的位点。 BRWD1差异监管超过7000 基因，反映了增殖和诱导分化程序。值得注意的是，BRWD1协调复制 MYC和MYC靶基因。此外，在人类中，BRWD1突变与 低磁性血症。这些数据指标表明，在小鼠和人类中，BRWD1都是主要的编排者 与TF网络合作以驱动B细胞晚期开发的增强器可访问性。 BRWD1首先在小型B细胞中表达。但是，幼稚的卵泡（FO）中BRWD1表达更高 在生发中心（GC）黑暗区（DZ）细胞中。在GC DZ和光中表达不同表达的基因的分析 与小型B细胞中BRWD1不同调节的区域（LZ）细胞（LZ）细胞预测BRWD1 压制DZ程序并诱导LZ程序。此外，鉴别基因组的结合 DZ和LZ细胞中的BRWD1在每个GC子集中提出不同的功能。免疫BRWD1FL/FL X AICDA（AID） - CRE小鼠表明，正常抗体同型切换和亲和力成熟需要BRWD1。 从组织学上讲，GC在大小和数量上都大大降低，并且缺乏卵泡树突状细胞。所以， 我们假设BRWD1调节表观遗传状态和增强剂景观对于DZ的协调至关重要 LZ转录程序和GC功能。我们将在以下具体目的中检验这一假设： 目标1。确定正常生发中心反应的转录调节。 目标2。确定BRWD1在自适应免疫组织化学中的作用。 目标3。确定BRWD1在启动GC响应和维持FO B细胞身份方面的作用。！