Distinct Functional Outcomes of BCR/TLR7 and BCR/TLR9 Co-engagement

BCR/TLR7 和 BCR/TLR9 共同参与的不同功能结果

基本信息

批准号：
9033830
负责人：
Ann Marshak-Rothstein
金额：
$ 51.36万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-12 至 2020-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9033830
关键词：
Accounting Adverse effects American Animal Disease Models Antibodies Antigen-Antibody Complex Autoantibodies Autoantigens Autoimmune Process Autophagosome B cell repertoire B-Cell Activation B-Lymphocytes Biochemical Biological Assay Bone Marrow CRISPR/Cas technology Cells Cessation of life ChIP-seq Chimera organism Clinical Computer Analysis Confocal Microscopy DNA Data Defect Dendritic Cells Dependence Development Diagnosis Disease Disease Progression Etiology Feedback Future Gene Expression Profile Gene Targeting Genetic Genomics Goals Health IRF4 gene Immunosuppressive Agents In Vitro Inbred BALB C Mice Inbred MRL lpr Mice Indium Lead Life Ligands Longevity Lupus Mediating MicroRNAs Modeling Molecular Monitor Mus Mutation Nature Nucleic Acid Binding Onset of illness PTPRC gene Pathway interactions Patients Pattern Pattern recognition receptor Phenotype Phosphorylation Phosphorylation Site Plasma Cells Plasmablast Play Population Post-Translational Protein Processing Pristane Process Production RNA Regimen Regulation Regulator Genes Risk Factors Role Scheme Source Staging Stem cells Surface Symptoms System Systemic Lupus Erythematosus TLR7 gene Techniques Technology Testing Therapeutic Toll-like receptors Transgenic Organisms Variant anti-IgM autoreactive B cell base cell type cohort design ds-DNA functional outcomes in vivo in vivo Model insight interest macrophage mouse model neutrophil novel protective effect purge receptor reconstitution response small molecule systemic autoimmune disease trafficking transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): SLE is a devastating systemic autoimmune disease of unknown etiology that presents with a diverse array of clinical symptoms and afflicts over 1.5 million Americans. Numerous risk factors have been identified, and it is generally accepted that SLE can result from a spectrum of immunoregulatory defects. Current treatments can involve immunosuppressive regimens associated with debilitating adverse side effects. Given the heterogeneous nature of the disease, it is remarkable that nucleic acid binding Toll-like receptors TLR7 and TLR9 has been found to play a critical role in the production of autoantibodies and disease development in all animal models of disease, and may prove to be critical targets for future therapies. Intriguingly, TLR9 appears to play both a protective and disease promoting role in the disease onset and progression - TLR9 is required for the production of autoantibodies against DNA-associated autoantigens but, quite inexplicably, TLR9-deficient autoimmune-prone mice develop much more severe clinical disease than their TLR9- sufficient counterparts. Our recent studies have now demonstrated a unique role for TLR9 in purging the autoreactive B cell repertoire - B cells activated by BCR/TLR9 engagement first undergo several rounds of division and then undergo a quite profound post-proliferative death. By contrast, BCR/TLR7 activated B cells survive and readily convert to plasmablasts. The goals of this project are to gain a better understanding of: (1) where in the course of disease TLR9 can mediate this protective effect; (2) when in the course of disease TLRs play a key role in B cell activation; (3) whether TLR9 mediates a suppressive role in any other cell types; (4) what unique features of the transcriptional networks downstream of BCR/TLR9 vs BCR/TLR7 co- engagement account for the distinct functional outcomes of these activation schemes, (5) how TLR9 and TLR7 contribute to autophagosome formation, and (6) how specific Unc93b post translational modifications preferentially promote TLR9 vs TLR7 activation in primary B cells, dendritic cells and macrophages. Our studies are based on a unique experimental system in which B cells are activated by physiologically relevant autoantigen immune complexes (IC), by using either IgG2a-specific transgenic B cells or by using a novel anti-IgM/IC dual variable domain polyclonal targeting antibody. ICs that incorporate both DNA and RNA will be used to parse the unique features of BCR/TLR9 and BCR/TLR7 activation pathways by varying the genetic background (TLR7- or TLR9-deficient) of the responder B cell populations. Advanced genomic and computational analyses will be used to elucidate gene regulatory networks. A highly efficient lentiviral retrogenesis strategy will be used to rapidly screen Unc93b variants for their capacity to support TLR9 vs TLR7 driven responses in specific cell types. These studies should provide important insights regarding the best strategies for designing effect TLR-based therapeutics for the treatment of patients diagnosed with SLE and other related systemic autoimmune diseases.

描述（由适用提供）：SLE是一种毁灭性的系统性自身免疫性病因疾病，呈现出一系列临床症状和150万美国人的临床症状和折磨。已经确定了许多危险因素，并且普遍认为SLE可能是由一系列免疫调节缺陷造成的。当前的治疗可能涉及与广告副作用相关的免疫抑制方案。鉴于该疾病的异质性质，已经发现，核酸结合Toll样受体TLR7和TLR9在所有疾病的自身抗体和疾病发展中起着至关重要的作用，这是值得注意的，并且可能证明是对未来疗法的关键靶标。有趣的是，TLR9似乎在疾病发作和进展中扮演疾病和促进疾病的作用-TLR9是针对与DNA相关的自身抗原的自身抗体所必需的，但是，TLR9缺乏性自身免疫性抗气的小鼠比起他们的TLR9严重临床疾病比他们的TLR9-STLR9-SERTLR9-SOUDCONT STLR9-SOUDCHS STHS-SERTICE COUNTER COUNTRASS COUNTRASS COUNTER SEFTICE COUNTER SEFTICE。我们最近的研究现在表明，TLR9在清除自动反应性B细胞库中 - BCR/TLR9互动激活的B细胞首先要进行几轮分裂，然后经历了相当深刻的后增生后死亡。相比之下，BCR/TLR7激活的B细胞存活并容易转化为浆膜。该项目的目标是更好地了解：（1）在疾病过程中TLR9可以介导这种保护作用；（2）在疾病过程中，TLR在B细胞激活中起关键作用；（3）TLR9是否在任何其他细胞类型中介导了抑制作用； (4) what unique features of the transcriptional networks downstream of BCR/TLR9 vs BCR/TLR7 co-engagement account for the distinct functional outcomes of these activation schemes, (5) how TLR9 and TLR7 contribute to autophagosome formation, and (6) how specific Unc93b post translatedal modifications preferentially promote TLR9 vs TLR7 activation in primary B cells,树突状细胞和巨噬细胞。我们的研究基于一个独特的实验系统，在该系统中，通过使用IgG2A特异性转基因B细胞或使用新型的抗IGM/IC Dualiable可变域多克隆靶靶抗体，通过使用物理相关的自身抗原免疫复合物（IC）激活B细胞。通过改变响应者B细胞群体的遗传背景（TLR7-或TLR9缺陷），将使用融合DNA和RNA的IC来解析BCR/TLR9和BCR/TLR7激活途径的独特特征。晚期基因组和计算分析将用于阐明基因调节网络。高效的慢病毒回培养策略将用于快速筛选UNC93B变体，以支持特定细胞类型中TLR9与TLR7驱动反应的能力。这些研究应提供有关设计基于TLR的最佳策略的重要见解，用于治疗被诊断为SLE和其他相关系统性自身免疫性疾病的患者。