Targeting follicular helper CD4 T cells in SLE

靶向 SLE 中的滤泡辅助 CD4 T 细胞

• 批准号: 10667605
• 负责人: Laurence Morel
• 金额: $ 46.5万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667605
• 关键词: Address; Adoptive Transfer; Affinity; Antibodies; Antigens; Autoantibodies; Autoimmune; Autoimmunity; B cell differentiation; B-Cell Activation; B-Lymphocytes; CD4 Positive T Lymphocytes; Cells; Cellular Metabolic Process; Chronic; Deoxyglucose; Disease; Disease model; Energy-Generating Resources; Enzymes; Explosion; FRAP1 gene; Frequencies; Funding; Genetic; Glucose; Glutamine; Glycolysis; Glycolysis Inhibition; Goals; Hypoxia; Immune; Immunity; Immunization; Immunoglobulin Class Switching; Immunoglobulin G; Immunophenotyping; Influenza; Knowledge; Lupus; Metabolic; Metabolism; Modeling; Mus; Norleucine; Oxidative Phosphorylation; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Population; Production; Program Sustainability; Proliferating; Proteins; Publishing; Role; Signal Transduction; Structure of germinal center of lymph node; Systemic Lupus Erythematosus; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Translating; autoreactive B cell; autoreactivity; cell type; ds-DNA; etomoxir; experimental study; fatty acid oxidation; genetic approach; in vivo; inhibitor; lupus prone mice; metabolomics; mouse model; pathogen; pharmacologic; preservation; response; single-cell RNA sequencing; therapeutic target; transcriptomics

Project Summary/Abstract High affinity class-switched autoantibodies (autoAb) are the major pathogenic effector molecules in lupus. They are generated by B cells that differentiate through either the germinal center (GC) or the extrafollicular (EF) pathways. The frequency of follicular helper (Tfh) cells and EF helper (eTf) CD4+ T cells are expanded in lupus in direct correlation with disease activity. The EF pathway has been best characterized in the MRL/lpr lupus- prone mice with AM14 BCR Tg B cells, which have been used as a model of autoreactive B cells activated through dual BCR and TLR9 signals. Overall, the GC and EF pathways are crucial nodes of lupus pathogenesis and lupus-prone mice present validated models for mechanistic studies of these pathways. We and others have shown that the metabolism of immune cells was altered in lupus patients and mice, and that some of these alterations offer therapeutic targets. Relatively little is known on the metabolic programs that sustain either autoreactive or immunization-induced (Imm-) GC B cells and Tfh cells, and nothing is known about the metabolism of EF activation. We have shown during the first cycle of funding that the production of anti-dsDNA IgG but not Imm-Abs, was glucose-dependent. In contrast, both autoAbs and Imm-Abs were eliminated by the inhibition of glutaminolysis. These results obtained with bulk polyclonal populations treated with metabolic inhibitors his led us to hypothesize that autoAbs and Imm-Abs have different metabolic requirements through the GC stage, which could be ultimately translated in their selective targeting. In this competitive renewal, we propose to use auto- and Imm-Ig-specific B and T cells, as well as GC B and Tact-specific deletions of metabolic enzymes to determine the intrinsic metabolic requirements of autoreactive and Imm-GC B and Tfh cells at the antigen- and cell-specific levels. We also propose to address the metabolic requirements of EF activation of autoreactive B cells. Using unique mouse models and a combination of scRNA-Seq and deep metabolomic analyses, we propose three specific aims to determine the requirements in glucose (1), glutamine (2), and FAO (3) for autoAb vs. Imm-Ab production, and to identify the corresponding mechanisms. These experiments will help to define the specific metabolic requirements of the B and CD4+ T cells that participate in the production of lupus autoAbs. This knowledge may ultimately identify means to eliminate autoAb production while preserving protective humoral autoimmunity.

项目概要/摘要 高亲和力类别转换自身抗体（autoAb）是狼疮的主要致病效应分子。他们 由通过生发中心 (GC) 或滤泡外 (EF) 分化的 B 细胞产生 途径。狼疮中滤泡辅助 (Tfh) 细胞和 EF 辅助 (eTf) CD4+ T 细胞的频率增加 与疾病活动度直接相关。 EF 途径在 MRL/lpr 狼疮中得到了最好的表征 具有 AM14 BCR Tg B 细胞的俯卧小鼠，该细胞已被用作激活的自身反应性 B 细胞的模型 通过双 BCR 和 TLR9 信号。总体而言，GC和EF途径是狼疮发病机制的关键节点 和易患狼疮的小鼠为这些途径的机制研究提供了经过验证的模型。我们和其他人有 研究表明，狼疮患者和小鼠的免疫细胞代谢发生了改变，其中一些 改变提供了治疗目标。对于维持两者的代谢程序知之甚少 自身反应性或免疫诱导的 (Imm-) GC B 细胞和 Tfh 细胞，目前尚不清楚 EF激活的代谢。我们在第一个资助周期中表明，抗 dsDNA 的生产 IgG 但不是 Imm-Abs，是葡萄糖依赖性的。相比之下，autoAbs 和 Imm-Abs 都被消除了。 抑制谷氨酰胺分解。这些结果是通过代谢处理的大量多克隆群体获得的 抑制剂使我们假设 autoAbs 和 Imm-Abs 通过以下方式具有不同的代谢需求： GC 阶段，这最终可以转化为选择性靶向。在这场竞争更新中，我们 建议使用自身和 Imm-Ig 特异性 B 和 T 细胞，以及代谢的 GC B 和 Tact 特异性删除 酶以确定自身反应性和 Imm-GC B 和 Tfh 细胞的内在代谢需求 抗原和细胞特异性水平。我们还建议解决 EF 激活的代谢要求 自身反应性 B 细胞。使用独特的小鼠模型以及 scRNA-Seq 和深度代谢组学的组合 通过分析，我们提出了三个具体目标来确定葡萄糖 (1)、谷氨酰胺 (2) 和 FAO (3) 比较 autoAb 与 Imm-Ab 的生产，并确定相应的机制。这些 实验将有助于确定参与参与的 B 和 CD4+ T 细胞的特定代谢需求 狼疮自身抗体的产生。这些知识可能最终确定消除自身抗体产生的方法 同时保留保护性体液自身免疫。