Cytokine-mediated B-cell development in lupus

细胞因子介导的狼疮 B 细胞发育

• 批准号: 10584137
• 负责人: John D Mountz
• 金额: --
• 依托单位: BIRMINGHAM VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10584137
• 关键词: Autoantibodies; Autoantigens; Autoimmune Diseases; B-Cell Activation; B-Cell Antigen Receptor; B-Cell Development; B-Lymphocytes; BACH2 gene; Beta Cell; Cell Cycle; Cell Surface Proteins; Cell surface; Cells; Characteristics; Clinical; Competence; Coupled; Data; Dedications; Defect; Development; Differentiation Antigens; Disease; Disease remission; Down-Regulation; Endosomes; Environmental Risk Factor; Equilibrium; Equipment; Evaluation; Event; Exhibits; Failure; Flow Cytometry; Gene Expression Profiling; Genes; Genetic Transcription; Goals; Human; IFNAR1 gene; ITGAX gene; Immune; Immune System Diseases; Immunoglobulin D; Immunoglobulin M; Immunologic Factors; Immunologics; In Vitro; Interferon Type I; Interferon Type II; Interferons; Interleukin 4 Receptor; Interleukin-4; Knock-in Mouse; Knock-out; Laboratories; Lupus; Maintenance; Mature B-Lymphocyte; Measures; Mediating; Medical center; Modeling; Mus; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Phenotype; Plasma Cells; Plasmablast; Prognosis; Proteins; Receptor Signaling; Rest; Role; Series; Signal Transduction; Sorting; Structure of germinal center of lymph node; Systemic Lupus Erythematosus; TLR7 gene; Techniques; Testing; Therapeutic; anergy; autoreactive B cell; bioinformatics pipeline; cytokine; diagnostic strategy; disease phenotype; experimental study; high dimensionality; immunoglobulin D receptor; imprint; improved; in vivo; innovation; lupus prone mice; mouse model; novel; novel diagnostics; novel therapeutic intervention; peripheral tolerance; prevent; programs; recruit; response; single cell analysis; therapeutic target; transcription factor; transcriptome; transcriptomics; type I interferon receptor; uptake

The overall goal of this project is to determine if the IKAROS transcription factor regulates a program that maintains IL-4 receptor (IL-4R)-mediated B-cell quiescence to ribonuclear protein (RNP) autoantigens (autoAgs). We propose that B cell activation through the type I interferon (IFN)/TLR7 activation pathway is inhibited by this B-cell quiescence program. We also propose that failure to maintain B cell quiescence is associated with the development of RNP autoantibody in systemic lupus erythematosus (SLE). Our preliminary data suggest that in both lupus mice and in human SLE, multiple immune disease phenotypes can be initiated by loss of quiescence at the earliest stage of B cell development; that is, the transitional B cell. The key forces that promote quiescence is an IKAROS (or IKZF1)-based program that is sustained through IL-4R signaling on IgD+CD23+ B cells. The major opposing program that promotes loss of quiescence is mediated through the type I interferon receptor (IFNAR) signaling, assembly and signaling competency of the TLR7 pathway, and development of B cells that can produce autoantibodies. We hypothesize that early-stage loss of quiescence leads to a series of B cell development defects after the transitional stage involving progression from a resting naïve to an activated naïve predominance and then to development of the pathogenic CD11c+Tbet+IgD−CD27− double negative 2 (DN2) or germinal center (GC) B cells that ultimately lead to the development of RNP autoantibody producing plasmablasts and plasma cells (PB/PC). In Aim 1, we will use several knockout and knock-in mouse strategies in lupus prone mice to determine if type I IFN and IL-4 act at the Tr B cells to regulate autoreactive B cell development and survival. The specific effects of Ikaros in modulating type I IFN and IL-4 signaling-mediated RNP-reactive B-cell quiescence at the Tr and naïve stage will also be analyzed. In Aim 2, we will determine if SLE patients exhibit a loss of IL-4R/IKZF1-mediated B cell quiescence program starting at the Tr stage of B-cell development. We will determine if the loss of this pathway disrupts B-cell tolerance to type I IFN and TLR7 stimulation, leading to the development of RNP- reactive DN2 B cells and PB/PC. The innovative scientific basis of this proposal will be its ability to interpret the broad spectrum of immune and disease characteristics of SLE by understanding the key initiating events of loss of B cell quiescence at the transitional stage. B cells from mouse models of lupus and from SLE subjects will be characterized by high dimension flow cytometry and transcriptomics analyses for well-established cell surface protein antigen markers that define B cell development at all major developmental stages. We have established a team of coordinators at the BVAMC to facilitate recruitment and have acquired numbers of SLE subjects necessary for definitive and statistically meaningful results. A state-of-the-art immunologic laboratory and BVAMC flow cytometry facility that includes the latest FACS analysis and sorting equipment, as well as a dedicated BVAMC 10X single-cell analysis facility, and analytical pipeline for bioinformatics analysis of the results are established. Significance: The successful conclusion of these experiments will be the development of new diagnostic and therapeutic approaches. From a diagnostic approach, the key defining phenotype of quiescent versus non-quiescent B cells can be economically and easily measured using standard clinical laboratory flow cytometry equipment with a relatively low number of cell surface (such as IL-4R or IFNAR1) or intracellular factors (such as intracellular IFNβ, TLR7, or IKAROS). This could be extended to analyze trajectories of abnormal B cell development that result from the loss of B cell quiescence at the earliest stages to understand therapeutics that may be beneficial in subjects who have acquired SLE in the past. At the very minimum, such approaches should enable the studies and types of treatments that will promote the maintenance of B cell quiescence and in effect, force SLE into a state of remission which can be maintained once the causes of the immune or environmental factors that disrupt such remission are defined.

该项目的总体目标是确定 IKAROS 转录因子是否调节以下程序： 维持 IL-4 受体 (IL-4R) 介导的 B 细胞对核糖核蛋白 (RNP) 自身抗原的静止 (autoAgs)。我们提出通过 I 型干扰素 (IFN)/TLR7 激活途径激活 B 细胞。 我们还提出，无法维持 B 细胞静止是由这种 B 细胞静止程序抑制的。 与系统性红斑狼疮 (SLE) 中 RNP 自身抗体的发展有关。 初步数据表明，在狼疮小鼠和人类 SLE 中，多种免疫疾病表型可以 由 B 细胞发育的最早阶段（即过渡 B 细胞）失去静止而引发。 促进静止的关键力量是基于 IKAROS（或 IKZF1）的程序，该程序通过 IL-4R 维持 IgD+CD23+ B 细胞上的信号传导是促进静止状态丧失的主要相反程序的介导。 通过 TLR7 的 I 型干扰素受体 (IFNAR) 信号传导、组装和信号传导能力 我们重新发现了早期的损失。 在涉及进展的过渡阶段后，静止状态会导致一系列 B 细胞发育缺陷 从静止的幼稚到激活的幼稚优势，然后到致病性的发展 CD11c+Tbet+IgD−CD27− 双阴性 2 (DN2) 或生发中心 (GC) B 细胞，最终导致 开发产生 RNP 自身抗体的浆母细胞和浆细胞 (PB/PC) 在目标 1 中，我们将使用。 狼疮易感小鼠中的几种敲除和敲入小鼠策略，以确定 I 型 IFN 和 IL-4 是否作用于 Tr B 细胞调节自身反应性 B 细胞发育和存活 Ikaros 的具体作用。 调节 I 型 IFN 和 IL-4 信号传导介导的 RNP 反应性 B 细胞在 Tr 和幼稚阶段的静止 在目标 2 中，我们还将确定 SLE 患者是否表现出 IL-4R/IKZF1 介导的 B 细胞缺失。 静止程序从 B 细胞发育的 Tr 阶段开始，我们将确定该程序是否丢失。 途径破坏 B 细胞对 I 型 IFN 和 TLR7 刺激的耐受性，导致 RNP-的发展 该提案的创新科学基础是其解释能力。 通过了解 SLE 的关键起始事件，了解 SLE 的广泛免疫和疾病特征 狼疮小鼠模型和 SLE 受试者的 B 细胞在过渡阶段失去静止状态。 将通过高维流式细胞术和对成熟细胞的转录组学分析来表征 我们拥有定义 B 细胞发育各个主要阶段的表面蛋白抗原标记。 在 BVAMC 建立了一个协调员团队以促进招募并已获得大量 SLE 获得明确且具有技术意义的结果所需的受试者。最先进的免疫学实验室。 和 BVAMC 流式细胞仪设施，包括最新的 FACS 分析和分选设备，以及 专用 BVAMC 10X 单细胞分析设施以及用于生物信息学分析的分析管道 结果成立 意义：这些实验的成功结论将是发展。 新的诊断和治疗方法的关键定义表型。 使用标准临床可以经济且轻松地测量静态 B 细胞与非静态 B 细胞 细胞表面数量相对较少的实验室流式细胞仪设备（例如 IL-4R 或 IFNAR1）或 细胞内因子（例如细胞内 IFNβ、TLR7 或 IKAROS）。 由于 B 细胞在最早阶段丧失静止状态而导致的异常 B 细胞发育轨迹 了解可能对过去患有 SLE 的受试者有益的治疗方法。 至少，这些方法应该能够促进研究和治疗类型的发展，从而促进 维持 B 细胞静止，实际上迫使 SLE 进入可维持的缓解状态 一旦确定了破坏这种缓解的免疫或环境因素的原因。