Epigenetic downregulation of the antibody and autoantibody response

抗体和自身抗体反应的表观遗传下调

基本信息

批准号：
9205214
负责人：
Paolo Casali
金额：
$ 47.32万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
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：
2014-02-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9205214
关键词：
3&apos Untranslated Regions Address Antibodies Antibody Affinity Antibody Response Antibody Suppression Antigens Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Autoimmunity B-Lymphocytes Bacteria Biochemistry Bioinformatics Biological Process Butyrates Cell physiology Cellular biology Chromatin Cytokine Receptors DNA Methylation Data Down-Regulation Epigenetic Process FDA approved Future Gene Silencing Gene Targeting Generations Genes Genetic Genetic Predisposition to Disease Genetic Recombination Genetic Transcription Histone Acetylation Histone Deacetylase Histone Deacetylase Inhibitor Histones Human Immunoglobulin Class Switching Immunoglobulin G Immunoglobulin Somatic Hypermutation Immunoglobulins In Vitro Inbred BALB C Mice Injury Knock-in Knock-in Mouse Knockout Mice Lupus Maps Mediating Methylation MicroRNAs Modification Molecular Molecular Biology Mus Mutate Nuclear Nuclear Antigens Organ PRDM1 gene Pathogenicity Pathway interactions Patients Penetrance Plasma Cells Porifera Post-Translational Protein Processing Prevention Process Production Regulation Research Role SLEB1 gene SLEB2 gene SLEB3 gene Signal Transduction Site Symptoms Systemic Lupus Erythematosus T-Independent Antigens TNFRSF5 gene Testing Therapeutic Tissues Untranslated RNA Untranslated Regions Up-Regulation Valproic Acid Virus Vorinostat chemical property crosslinking and immunoprecipitation sequencing disorder prevention ds-DNA histone modification immune function immunoregulation in vivo inhibitor/antagonist innovation insight killings lupus prone mice microbial microorganism antigen mutant neoplastic cell novel therapeutics pathogen plasma cell differentiation prevent programs public health relevance response sound targeted treatment tool

3&apos Untranslated Regions Address Antibodies Antibody Affinity Antibody Response Antibody Suppression Antigens Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Autoimmunity B-Lymphocytes Bacteria Biochemistry Bioinformatics Biological Process Butyrates Cell physiology Cellular biology Chromatin Cytokine Receptors DNA Methylation Data Down-Regulation Epigenetic Process FDA approved Future Gene Silencing Gene Targeting Generations Genes Genetic Genetic Predisposition to Disease Genetic Recombination Genetic Transcription Histone Acetylation Histone Deacetylase Histone Deacetylase Inhibitor Histones Human Immunoglobulin Class Switching Immunoglobulin G Immunoglobulin Somatic Hypermutation Immunoglobulins In Vitro Inbred BALB C Mice Injury Knock-in Knock-in Mouse Knockout Mice Lupus Maps Mediating Methylation MicroRNAs Modification Molecular Molecular Biology Mus Mutate Nuclear Nuclear Antigens Organ PRDM1 gene Pathogenicity Pathway interactions Patients Penetrance Plasma Cells Porifera Post-Translational Protein Processing Prevention Process Production Regulation Research Role SLEB1 gene SLEB2 gene SLEB3 gene Signal Transduction Site Symptoms Systemic Lupus Erythematosus T-Independent Antigens TNFRSF5 gene Testing Therapeutic Tissues Untranslated RNA Untranslated Regions Up-Regulation Valproic Acid Virus Vorinostat chemical property crosslinking and immunoprecipitation sequencing disorder prevention ds-DNA histone modification immune function immunoregulation in vivo inhibitor/antagonist innovation insight killings lupus prone mice microbial microorganism antigen mutant neoplastic cell novel therapeutics pathogen plasma cell differentiation prevent programs public health relevance response sound targeted treatment tool

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项目摘要

DESCRIPTION (provided by applicant): Epigenetic downregulation of the antibody and autoantibody response Epigenetic marks include DNA methylation, histone modifications and microRNAs. As we have contended, these "interact" with genetic programs to regulate B cell functions, such as class switch DNA recombination (CSR), somatic hypermutation (SHM) and plasma cell differentiation, thereby informing the antibody response. Epigenetic dysregulation can result in aberrant antibody responses to exogenous antigens, such as those on viruses and bacteria, or self-antigens, such as chromatin, histones and dsDNA in lupus. We hypothesize that the epigenetic modulators histone deacetylase (HDAC) inhibitors (HDIs) inhibit the B cell intrinsic functions CSR/SHM and plasma cell differentiation, thereby blunting antibody and autoantibody responses. We further argue that HDIs inhibit these B cell functions by upregulating selected microRNAs, including miR-155, miR-181b and miR-361 to downregulate AID (Aicda gene, critical for CSR/SHM), as well as miR-23b, miR-30a and miR-125 to downregulate Blimp1 (Prdm1 gene, critical for plasma cell differentiation). Our hypotheses uniquely focus on B cells and are supported by our compelling data using purified human and mouse B cells in vitro, and in vivo in normal mice responding to T-dependent and T- independent antigens, and prevention of disease and "treatment" of lupus-prone mice by HDI. Our strengths in B cell biology, molecular CSR/SHM mechanisms and autoimmunity, and our cutting-edge epigenetic tools and approaches make us uniquely poised to test our hypotheses. We will: (Aim 1) analyze the impact of HDIs (valproic acid, butyrate and SAHA) on CSR/SHM and plasma cell differentiation, and on specific T-dependent and T-independent antibody responses in normal C57BL/6 and Balb/c mice and autoantibody responses in lupus-prone MRL/Faslpr/lpr and B6.NZM/Sle1.Sle2.Sle3 mice; (Aim 2) use molecular biology, biochemistry and high-throughput/bioinformatics tools to analyze HDI-mediated upregulation of miR-155, miR-181b, miR-361, miR-23b, miR-30a and miR-125b in B cells through enhanced histone acetylation and transcription of the microRNA "host genes", address downregulation of AID and Blimp1 by these microRNAs, and construct microRNA in vivo targeting maps of Aicda and Prdm1 3'UTRs using Ago HITS-CLIP; (Aim 3) prove the critical role of B cell microRNAs in mediating HDI suppression of antibody and autoantibody responses using a three-prong integrated in vivo approach involving construction of new knockin mice lacking specific microRNA targeting sites in Aicda or Prdm1 3'UTR, B cell conditional Dicer or Drosha KO mice, and mice expressing "sponge" inhibitors specific for miR-155, miR-181b and miR-361 (targeting Aicda) and/or for miR-23b, miR-30a and miR-125b (targeting Prdm1) in B cells. Our proposal is highly innovative and exquisitely translational. It will provide mechanistic insights and future directions in epigenetics and immunoregulation, including the critical role of B cell microRNAs in antibody/autoantibody responses and epigenetic tools as new therapeutics in autoimmunity.

描述（由申请人提供）：抗体和自身抗体反应表观遗传标记的表观遗传下调包括DNA甲基化，组蛋白修饰和microRNA。正如我们争辩的那样，这些与遗传程序的“相互作用”以调节B细胞功能，例如类开关DNA重组（CSR），体外超突变（SHM）和浆细胞分化，从而告知抗体反应。表观遗传失调会导致对外源抗原的异常抗体反应，例如病毒和细菌的抗原，或狼疮中的自我抗原，例如染色质，组蛋白和dsDNA。我们假设表观遗传调节剂组蛋白脱乙酰基酶（HDAC）抑制剂（HDIS）抑制B细胞固有函数CSR/SHM和血浆细胞分化，从而使抗体和自身抗体反应钝化。 We further argue that HDIs inhibit these B cell functions by upregulating selected microRNAs, including miR-155, miR-181b and miR-361 to downregulate AID (Aicda gene, critical for CSR/SHM), as well as miR-23b, miR-30a and miR-125 to downregulate Blimp1 (Prdm1 gene, critical for plasma cell differentiation).我们的假设独特地集中在B细胞上，并在体外使用纯化的人和小鼠B细胞的引人注目的数据以及对T依赖性和T独立抗原作出反应的正常小鼠的体内以及预防HDI的狼疮prolon小鼠的疾病和“治疗”。我们在B细胞生物学，分子CSR/SHM机制和自身免疫性以及最先进的表观遗传学工具和方法方面的优势使我们能够独特地测试我们的假设。我们将：（目标1）分析HDI（丙丙酸，丁酸酯和saha）对CSR/SHM和浆细胞分化的影响，以及在正常C57BL/6和BALB/CILB/CILB/FPAS-PRONE/FASPR/FASPR/lpr的特定T依赖性和T依赖性T依赖性和T-独立抗体响应中。小鼠；（目标2）使用分子生物学，生物化学和高通量/生物信息学工具来分析HDI介导的miR-155，miR-181b，miR-361，miR-361，miR-23b，miR-30a，miR-30a和miR-125b在B细胞中通过增强的组型乙酰基和转录的MicroRERN和Bromer的Microrn and Infrient and Mirstription和Mirstription of Microrn and of Microlas of Microrn and of Microrn and of Microrn，“使用AGO hits-clip构造MicroRNA在体内靶向AICDA和PRDM1 3'UTR的地图；（目标3）证明了B细胞microRNA在介导HDI抑制抗体和自身抗体反应中的关键作用，该反应使用三孔整合在体内方法中，涉及构建AICDA或PRDM1 3'UTR中缺乏特定microRNA靶向位点的新敲蛋白小鼠，B细胞doser tement diCer或Drosha ko sexs and Drosha ko seme sons and temend sexpon ins and sexpon ko ko nifif nifing''in in in' miR-155，miR-181b和miR-361（靶向AICDA）和/或用于miR-23b，miR-30a和miR-125b（靶向PRDM1）。我们的建议具有高度创新性和精致的翻译。它将提供表观遗传学和免疫调节的机械见解和未来方向，包括B细胞microRNA在抗体/自身抗体反应中的关键作用，以及作为自身免疫中的新疗法的抗体/自身抗体反应和表观遗传学工具。