An Autoimmune center of excellence for the study of IgG4-related disease

研究 IgG4 相关疾病的自身免疫卓越中心

• 批准号: 8680709
• 负责人: SHIV Subramaniam PILLAI
• 金额: $ 71.29万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8680709
• 关键词: Antibodies; Antibody Repertoire; Antigens; Autoimmune Diseases; Autoimmune Process; B-Lymphocytes; Binding; CD4 Positive T Lymphocytes; Cell Surface Proteins; Cells; Cessation of life; Characteristics; Cicatrix; Clinical; Clonal Expansion; Clone Cells; Color; Cytometry; Development; Disease; Epigenetic Process; Expression Library; Fc Receptor; Fibrosis; Flow Cytometry; Gene Expression; Gene Expression Profiling; Genetic Predisposition to Disease; Genotype; Hamman-Rich syndrome; Human; IgG4; Immunology; Inflammatory; Knowledge; Lesion; Libraries; Light; MHC Class II Genes; Molecular; Organ; Pathogenesis; Pathway interactions; Patients; Peptides; Phlebitis; Pilot Projects; Plasma; Plasmablast; Property; Proteins; Quality of life; Reagent; Recombinant Proteins; Research Design; Role; Serum; Source; Steroids; Surface; Survivors; Swelling; Syndrome; System; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Therapeutic; Tissues; base; commensal microbes; exome sequencing; glycosylation; human monoclonal antibodies; interdisciplinary approach; liquid chromatography mass spectrometry; microbial community; multidisciplinary; novel; oral tissue; premature; protein expression; protein metabolite; response; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): In this project we will seek to better understand the pathogenesis of lgG4 related disease utilizing a multidisciplinary approach. Based on this knowledge we seek to influence the development of new therapies that may be applicable not just to this disease but to other autoimmune and fibrotic diseases as well. lgG4 related disease is a multisystem disorder characterized by tissue swelling/s, storiform fibrosis, obliterative phlebitis and high levels of plasma lgG4. While this disorder is likely to be autoimmune, an aberrant response to a pathogenic or commensal microbe has not been ruled out. In the principal proposal we will study in depth activated effector CD4+ T cell clones in subjects with lgG4-Related disease. We will examine the T cell receptor repertoire by Next Gen Sequencing, and obtain a detailed analysis of gene and protein expression in these clones in order to better understand how they are generated and how they function. We will perform single cell RNA seq and mass cytometry to understand the clonal origins of these effector T cells. Unique cell surface proteins found only in disease subjects will be identified. The mechanism by which these T cell clones influence fibrosis will be examined. The antibody repertoire of plasmablasts will also be identified, and novel disease related human monoclonal antibodies will be generated and the properties of clonally expanded plasmablasts will also be studied. Human ORFeome libraries will be interrogated with serum and disease related monoclonal lgG4 antibodies and disease specific antigens will be identified and used as tools to identify the antigenic peptides recognized by clonally expanded CD4+ T cells. In the Collaborative project studies will be performed to correlate oral, tissue and gut microbial communities with genetic susceptibility markers and subsets of patients with distinct functional characteristics of the clonal effector T cells in disease subjects. In the Pilot Project the glycosylation of lgG4, in subjects with active disease will be analyzed and its ability to influenc binding to different human Fc receptors will be examined. RELEVANCE: The proposed multidisciplinary studies are designed not only to benefit patients with lgG4-related disease, but to also benefit a host of other disorders including many other autoimmune diseases and other diseases including idiopathic pulmonary fibrosis. Principal Project: Studies on the Immunology of IgG4-related diseases Project Leader (PL): Shiv S. Pillai DESCRIPTION (as provided by applicant): lgG4-Related Disease is a multi-system disorder encompassing a host of previously described syndromes, all now recognized to be characterized by tumescent lesions, storiform fibrosis, obliterative phlebitis and large amounts of serum lgG4. Clinical improvement is seen with steroids in many subjects and B cell depletion is also clinically effective. Very little is known about the pathogenesis of this disorder or about th molecular and cellular basis for fibrosis in a host of apparently unrelated disorders. Studies will be performed to define T cell clonal expansions observed by Next Gen Sequencing approaches in subjects with this disease but with distinct organ involvements. Novel surface markers expressed only on effector T cell clones will be investigated as potential targets for therapy. Detailed interrogation of gene expression protein expression and metabolites will be performed by global as well as single cell RNAseq, multi-color flow cytometry, by Cytof mass cytometry, and liquid chromatography-mass spectrometry on clonally expanded T cells and will be conducted in order to understand pathways of potential therapeutic significance that contributes to the development of these T cells as well as to their effector functions in this fibrotic inflammatory disease. A possible role for lgG4 antibodies in this disordr will also be examined. Next Gen Sequencing as well as single cell cloning and sequencing strategies will be used to define plasmablast expansions and to establish specific antibody heavy-light chain pairs that may contribute to the disease. Reagents will be thus generated to identify specific antigens using human ORFeome expression libraries as source of antigen. Determining the B cell specific protein antigen and the use of recombinant proteins will be used to assist the identification of T cell specific peptides. Studies will also be performed on genetic susceptibility to lgG4-RD using Fluidigm based MHC class II genotyping and if indicated from the Immunochip analyses by Exome sequencing. Global comparisons of gene and protein expression will inform the need for epigenetic profiling studies. RELEVANCE: Autoimmune and inflammatory diseases that cause severe tissue scarring or fibrosis can cause premature death and seriously impair the quality of life for survivors. The proposed studies may help not only patients with lgG4-related disease, but also a host of other disorders including many other autoimmune diseases and other diseases including idiopathic pulmonary fibrosis.

描述（由申请人提供）：在这个项目中，我们将利用多学科方法更好地了解 lgG4 相关疾病的发病机制。基于这些知识，我们寻求影响新疗法的开发，这些新疗法不仅适用于这种疾病，还适用于其他自身免疫性疾病和纤维化疾病。 IgG4相关疾病是一种多系统疾病，其特征为组织肿胀、席纹状纤维化、闭塞性静脉炎和血浆IgG4水平高。虽然这种疾病可能是自身免疫性疾病，但尚未排除对致病或共生微生物的异常反应。在主要提案中，我们将深入研究患有 IgG4 相关疾病的受试者中激活的效应 CD4+ T 细胞克隆。我们将通过下一代测序检查 T 细胞受体库，并对这些克隆中的基因和蛋白质表达进行详细分析，以便更好地了解它们是如何产生的以及它们如何发挥作用。我们将进行单细胞 RNA 测序和质谱流式分析来了解这些细胞的克隆起源 效应T细胞。将鉴定仅在疾病受试者中发现的独特细胞表面蛋白。我们将研究这些 T 细胞克隆影响纤维化的机制。浆母细胞的抗体库也将被鉴定，并且将产生新的疾病相关的人单克隆抗体，并且还将研究克隆扩增的浆母细胞的特性。将用血清和疾病相关的单克隆 IgG4 抗体对人 ORFeome 文库进行询问，并且将鉴定疾病特异性抗原并用作鉴定由克隆扩增的 CD4+ T 细胞识别的抗原肽的工具。在合作项目中，将进行研究，将口腔、组织和肠道微生物群落与遗传易感性标记以及疾病受试者中具有克隆效应 T 细胞独特功能特征的患者亚群关联起来。在试点项目中，将分析患有活动性疾病的受试者中 IgG4 的糖基化，并检查其影响与不同人类 Fc 受体结合的能力。 相关性：拟议的多学科研究不仅旨在使 lgG4 相关疾病的患者受益，而且还使许多其他疾病受益，包括许多其他自身免疫性疾病和包括特发性肺纤维化在内的其他疾病。 主持项目：IgG4相关疾病的免疫学研究 项目负责人 (PL)：Shiv S. Pillai 描述（如申请人提供的）：IgG4 相关疾病是一种多系统疾病，包括许多先前描述的综合征，所有这些综合征现在都被认为以肿胀病变、席纹状纤维化、闭塞性静脉炎和大量 血清IgG4。在许多受试者中使用类固醇可以看到临床改善，并且 B 细胞去除在临床上也是有效的。对于这种疾病的发病机制或许多明显不相关的疾病中纤维化的分子和细胞基础知之甚少。研究将 进行以确定通过下一代测序方法在患有这种疾病但具有不同器官受累的受试者中观察到的 T 细胞克隆扩增。仅在效应 T 细胞克隆上表达的新型表面标记将作为潜在的治疗靶点进行研究。将通过全局和单细胞 RNAseq、多色流式细胞术、Cytof 质谱流式细胞术以及克隆扩增 T 细胞的液相色谱-质谱法对基因表达蛋白表达和代谢物进行详细询问，并进行以下操作：了解潜在治疗途径 有助于这些 T 细胞的发育及其在这种纤维化炎症疾病中的效应功能。还将检查 IgG4 抗体在这种疾病中的可能作用。下一代测序以及单细胞克隆和测序策略将用于定义浆母细胞扩增并建立可能导致疾病的特异性抗体重轻链对。因此将产生试剂以使用人ORFeome表达文库作为抗原来源来鉴定特定抗原。确定B细胞特异性蛋白抗原并使用重组蛋白将有助于辅助T细胞特异性肽的鉴定。 还将使用基于 Fluidigm 的 MHC II 类对 lgG4-RD 的遗传易感性进行研究 基因分型，以及外显子组测序的免疫芯片分析是否表明。基因和蛋白质表达的全局比较将告知表观遗传分析研究的需要。 相关性：导致严重组织疤痕或纤维化的自身免疫和炎症性疾病可能导致过早死亡并严重损害幸存者的生活质量。拟议的研究不仅可以帮助患有 lgG4 相关疾病的患者，还可以帮助治疗许多其他疾病，包括许多其他自身免疫性疾病和特发性肺纤维化等其他疾病。