Interferon Beta Initiated Development of Immunogenic T1 B cells in Lupus

干扰素 Beta 启动狼疮中免疫原性 T1 B 细胞的发育

基本信息

批准号：
10477180
负责人：
John D Mountz
金额：
--
依托单位：
BIRMINGHAM VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-10-01 至 2022-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10477180
关键词：
Antibodies Antigens Apoptotic Autoantibodies Autoantigens Autoimmune Diseases Autoimmunity Automobile Driving B-Cell Antigen Receptor B-Lymphocyte Subsets B-Lymphocytes Biological Response Modifier Therapy Bone Marrow Caring Cell Compartmentation Cell Maturation Cell Separation Cell Survival Cells Chimera organism Chronic Clinical Cluster Analysis Data Defect Development Disease Elements Environmental Risk Factor Event Exhibits FOXP3 gene Flow Cytometry Funding Gene Expression Gene Expression Profile Gene Expression Profiling Generations Genetic Predisposition to Disease Genetic Transcription Goals Grant IFNAR1 gene IRF3 gene Immunocompetent Immunoglobulin Idiotypes In Vitro Interferon Receptor Interferon Type I Interferon-alpha Interferon-beta Interferons Interleukin-10 Lupus Maintenance Mature B-Lymphocyte Mediating Modeling Molecular Mouse Strains Mus Nuclear Nucleic Acids Pathogenesis Pathogenicity Pathway interactions Patients Phenotype Play Population Process Production Reporter Resolution Role Signal Transduction Specificity Systemic Lupus Erythematosus T-Lymphocyte TLR7 gene Techniques Testing Therapeutic Transitional Cell Up-Regulation Work autocrine autoreactive B cell autoreactivity base cytokine design experimental study immunogenic imprint improved in vivo insight inter-individual variation migration mouse model novel precision medicine receptor-mediated signaling response single cell analysis therapeutic target tool transcription factor

项目摘要

Systemic lupus erythematosus (SLE) is a chronic debilitating disease that is characterized by high titers of autoantibodies with specificity for nuclear autoantigens. It is considered to arise from interactions between underlying genetic susceptibility and environmental factors, with different combinations resulting in inter- individual variations in disease manifestations and therapeutic responsiveness. Directed therapeutic targeting of specific pathways has not proven effective, which suggests that the current understanding of pathogenesis is incomplete. The goal of the proposed project is to test the hypothesis that the survival of immature autoreactive B cells, the preferential development of antinuclear autoreactive B cells and the status of mature autoreactive B cells (immunocompetent vs. anergic/tolerogenic) is decided during the transitional stage 1 (T1) of development and requires constitutive T1 B cell expression of interferon β (IFNβ). This paradigm shifting hypothesis is based on data generated using a combination of mixed-bone marrow chimeras, flow cytometry with tetramer or idiotype antibody selection, and high-throughput single-cell analysis to interrogate type I interferon (IFN) networks in B cell from patients with SLE and lupus-prone BXD2 mice. The data suggest that the currently known molecular and cellular aberrations are preceded by a single primary pathogenic event, i.e., production of IFNβ by T1 B cells. The data suggest that the survival of the very early T1 cells is dependent on endogenous expression of IFNβ. This subset of T1 B cells express endogenous IFNβ, which leads to their development into T1 B cells that produce IFNα. The ability of T1 B cells to escape negative B-cell receptor (BCR)-mediated selection is dictated by the type I IFN-determined responsiveness of this T1 subset to BCR-mediated signaling in combination with stimulation of TLR signaling (TLR7 or TLR9) by apoptotic debris. This preferentially permits escape of nucleic antigen-autoreactive B cells. We have further identified that the T1 B cell compartment contains distinct subsets of cells. As these included subsets with transcriptional profiles that parallel the phenotypes of mature and anergic suppressive regulatory (Breg) cells and their immunogenic counterparts that predominate in SLE, our new data suggest that this phenotypic switch in the mature B cells is imprinted during the T1 stage. These data also suggest that development of type I IFN network associated T1 B cells in BXD2 mice is associated with type I IFN-inducing transcription factors (IRF3 and IRF7) whereas induction of the Foxp3+ regulatory T1 B cells is associated with transcription factor, ID3. Notably, these data not only suggest suppression of precursors of T1-Breg is a novel pathogenic framework in SLE but also provide the tool, i.e., the gene expression signatures in sorted transitional B cells, that enable its analysis. We will test the critical elements of our overall hypothesis through three Specific Aims that will: (1) Distinguish the roles of the constitutive IFNβ signaling and the microenvironmental signals in driving the development of T1 B cells; (2) Identify if the initial T1 B cell transcriptional imprinting is maintained at the T2/MZP-MZ-FO stage of development; and (3) Determine if T1 B-cell endogenous IFNβ promotes a type I IFN network that suppresses the precursors of regulatory B cells at the T1 stage in SLE patients. The scientific premises of the proposed work include the parallels between transcriptional profiles in the T1 B cells and mature B cells and the segregation of type I IFN associated vs Breg associated transcriptional profiles in the T1 B cells. We have acquired reporter mouse strains for IFNβ, IL-10, Foxp3 and Id3 to interrogate discreet developmental stages within and derived from transitional T1 B subsets. The information gained will have a broad-based impact on the field and improve the care of VA patients with SLE by identifying strategies to improve the efficacy of targeted biologic therapies together with non- invasive precision medicine guided-approaches.

全身性红斑狼疮（SLE）是一种慢性衰弱的疾病，其特征是高滴度具有特异性的自身抗体。它被认为是由潜在的遗传敏感性和环境因素，不同的组合导致疾病表现和治疗反应性的个人变化。定向治疗靶向特定途径尚未证明有效，这表明当前对发病机理的理解是不完整的。拟议项目的目的是检验未成熟生存的假设自动反应性B细胞，抗核自身反应性B细胞的首选发展和成熟的状态自动反应性B细胞（免疫能力与厌食/耐受性）是在过渡阶段1（T1）中决定的发育和需要干扰素β（IFNβ）的组成型T1 B细胞表达。该范式转移假设基于使用混合骨髓的组合生成的数据嵌合体，具有四聚体或白痴型抗体选择的流式细胞仪以及高通量单细胞分析询问来自SLE和狼疮和狼疮的BXD2小鼠患者的B细胞中的I型干扰素（IFN）网络。这数据表明，当前已知的分子和细胞像差之前是单个主要的致病事件，即由T1 B细胞生产IFNβ。数据表明T1早期的生存细胞取决于IFNβ的内源性表达。 T1 B细胞的这一子集表达内源性IFNβ，这导致它们发展为产生IFNα的T1 B细胞。 T1 B细胞逃脱阴性的能力 B细胞接收器（BCR）介导的选择取决于该T1的I型IFN确定的响应能力子集至BCR介导的信号传导与刺激TLR信号传导（TLR7或TLR9）结合凋亡碎片。这优先允许逃脱核抗原运动性B细胞。我们还有更多确定T1 B细胞室包含不同的细胞子集。因为这些包括子集与成熟抑制性调节（Breg）细胞的表型相似的转录曲线他们的免疫原性对应物在SLE中以SLE为主导，我们的新数据表明这种表型开关在成熟的B细胞中，在T1阶段印刷。这些数据还表明，I型IFN的发展 BXD2小鼠中与网络相关的T1 B细胞与I型IFN诱导的转录因子有关（IRF3 和IRF7），而Foxp3+调节T1 B细胞的诱导与转录因子ID3有关。值得注意的是，这些数据不仅表明抑制T1-BREG的前体是一种新型的致病性 SLE中的框架，但还提供该工具，即，在分类的过渡B细胞中的基因表达签名，这可以分析。我们将通过三个特定目标来检验整体假设的关键要素这将是：（1）区分本构IFNβ信号传导和微环境信号的作用推动T1 B细胞的发展；（2）确定最初的T1 B细胞转录印记是否保持在T2/MZP-MZ-FO开发阶段；（3）确定T1 B细胞内源性IFNβ是否促进 I IFN网络可在SLE患者的T1阶段抑制调节B细胞的前体。这拟议工作的科学前提包括T1 B细胞中转录曲线之间的相似之处和成熟的B细胞以及I型IFN相关与Breg相关的转录曲线的分离 T1 B细胞。我们已经获得了用于IFNβ，IL-10，FOXP3和ID3的记者小鼠菌株以审问谨慎内部的发展阶段并源自过渡T1 B子集。获得的信息将对该领域产生广泛的影响并改善VA患者的护理通过确定策略来提高目标生物疗法的效率以及非 - 侵入性的精确药物指导。