B cell Autoimmunity in Human SLE

描述(申请人提供)：B细胞在许多人类自身免疫性疾病中起中心致病作用，包括系统性红斑狼疮(SLE)。理解和有效治疗人类自身免疫将需要深入了解导致B细胞耐受性崩溃的抗原、细胞和分子事件，以及通过产生效应细胞而导致急性临床疾病的下游事件，以及通过形成、维持和重新激活长期自身免疫记忆而导致慢性自身免疫的事件。埃默里自身免疫卓越中心(ACE)U19提出了一种综合方法，通过三个高度协同的项目和最先进技术的使用来阐明这些核心问题。主要项目(Sanz博士，Pi)将阐明幼稚细胞和记忆细胞在急性狼疮发作期间扩大的抗体分泌细胞的生成中的相对参与，导致这些扩大的抗原性驱动因素，以及不同类型狼疮自身抗体的细胞基础。协作议程(Boss博士，Pi)将使用转录和表观遗传学研究来绘制导致主要项目中涉及的细胞事件的信号通路。反过来，这些研究将提供一个科学和技术平台，Emory ACE U19将扩展到其他ACE中心，以类似地研究其他免疫细胞类型和人类自身免疫性疾病。最后，飞行员 项目(Dr.Jacob，Pi)将剖析导致人类长寿浆细胞存活的分子决定因素。 Emory ACE U19的科学使命将得到由Sanz博士领导的整合良好的管理核心的支持，Sanz博士作为Rochester ACE的PI已经担任了10多年的类似角色。Emory ACE还将为ACE指导委员会提供重要投入，通过U19协作议程和UMI支持的临床试验，积极促进制定网络的科学议程，并为机械研究的设计和实施做出贡献。 相关性：Emory ACE U19将对人类SLE中B细胞功能障碍的细胞和分子机制进行综合研究。通过合作项目，它将扩大研究其他免疫细胞和自身免疫性疾病的范围。预期的结果应该会大大提高我们诊断、治疗和预防自身免疫性疾病的能力。 主要项目：人类系统性红斑狼疮中的B细胞 项目负责人：伊格纳西奥·桑兹 描述：系统性红斑狼疮是一种全身性自身免疫性疾病，以严重的B细胞异常和多种自身抗体产生为特征。然而，尽管人类B细胞生物学领域取得了重大进展，但SLE中B细胞激活、多样化和扩增的确切抗原性和细胞机制仍然知之甚少。此外，疾病异质性、B细胞表型不精确以及缺乏高通量技术来确定驱动自身反应性B细胞和血清自身抗体产生和选择的抗原力，阻碍了对不同B细胞亚群在急性疾病加重期间相对参与的准确理解。在当前的ACE周期中，我们已经建立了解决这些问题所需的工具，包括多维流式细胞术；下一代测序(NGS)；大规模单细胞抗体生成；以及用于高密度免疫学、转录和临床数据综合分析的生物信息学平台。此外，我们还启动了与专家小组的合作，以研究B细胞和ASC表观遗传学(Boss博士，协作议程PI)和血清抗体蛋白质组学分析(Cheung博士；CST)。在Emory ACE U19的这一主要项目中，我们建议在这些成就的基础上，通过以下具体目标来剖析B细胞和ASC亚群的参与：目的1.利用NGS谱系分析和单细胞抗体产生来参与和抗原选择不同的B细胞亚群；目的2.短寿命和长寿命抗体分泌细胞对SLE血清自身抗体蛋白质组的相对贡献；以及目的3.SLE B细胞的表观遗传和转录控制与合作项目。拟议的工作与《合作议程》和试点项目高度协同，在提出的问题和试验方法方面具有高度的创新性。预期的结果对于我们理解人类自身免疫中作用于B细胞的致病机制以及设计更安全、更有效的治疗方法具有深远的意义。 相关性：该项目将提供对不同的B细胞和抗体分泌细胞在人类SLE自身抗体谱系和疾病过程中的贡献的关键洞察。与合作项目一起，它还将研究自身免疫B细胞的表观基因组和转录程序。

DESCRIPTION (provided by applicant): B cells play central pathogenic roles in many human autoimmune diseases including Systemic Lupus Erythematosus (SLE). Understanding and effectively treating human autoimmunity will require a deep understanding of the antigenic, cellular and molecular events that contribute to the breakdown of B cell tolerance and the downstream events leading to acute clinical disease through the generation of effector cells and to chronic autoimmunity through the formation, maintenance and reactivation of long-lived autoimmune memory. The Emory Autoimmunity Center of Excellence (ACE) U19 proposes an integrated approach to elucidating these central questions through three highly synergistic projects and the use of state-of-the-art technology. The Principal Project (Dr. Sanz, PI), will elucidate the relative participation of naive and memory cells in the generation of antibody secreting cells expanded during acute lupus flares, the antigenic drives responsible for these expansions and the cellular basis of different types of lupus autoantibodies. The Collaborative Agenda (Dr. Boss, PI), will use transcriptional and epigenetic studies to map the signaling pathways responsible for the cellular events addressed in the Principal Project. In turn, these studies will provide a scientific and technological platform that the Emory ACE U19 will extend to other ACE centers to similarly study other immune cell types and human autoimmune diseases. Finally, the Pilot Project (Dr. Jacob, PI), will dissect the molecular determinants responsible for the survival of human long-lived plasma cells. The scientific mission of the Emory ACE U19 will be supported by a well-integrated Administrative Core led by Dr. Sanz who has served in a similar role for more than 10 years as PI of the Rochester ACE. The Emory ACE will also provide major input into the ACE Steering Committee actively contributing to developing the scientific agenda of the network and to the design and implementation of mechanistic studies both through the U19 Collaborative Agenda and the UMI-supported clinical trials. RELEVANCE: The Emory ACE U19 will pursue an integrated approach to the cellular and molecular mechanisms of B cell dysfunction in Human SLE. Through the Collaborative Project, it will enlarge its reach to study other immune cells and autoimmune diseases. The expected results should greatly enhance our ability to diagnose, treat and prevent autoimmune diseases. Principal Project: B Cells in Human SLE Project Leader: Ignacio Sanz DESCRIPTION: SLE is a systemic autoimmune disease characterized by profound B cell abnormalities and multiple autoantibody production. However, despite major advances in the field of human B cell biology, the precise antigenic and cellular mechanisms that underlie the activation, diversification and expansion of B cells in SLE remain poorly understood. Moreover, a precise understanding of the relative participation of different B cell subsets during acute disease exacerbations has been hampered by disease heterogeneity, imprecise B cell phenotyping and the lack of high-throughput technologies needed to define the antigenic forces driving the generation and selection of autoreactive B cells and serum autoantibodies. During the current ACE cycle we have established the tools required to address these questions including multi-dimensional flow cytometry; next generation sequencing (NGS); large scale single cell antibody generation; and bioinformatics platforms for the integrative analysis of high-density immunological, transcriptional and clinical data. Moreover, we have initiated collaborations with expert groups for the study of B cell and ASC epigenetics (Dr. Boss, Collaborative Agenda PI) and for the analysis of serum antibody proteomics (Dr. Cheung; CST). In this Principal Project of the Emory ACE U19, we proposed to build on these accomplishments to dissect the participation of B cell and ASC subsets through the following specific aims: Aim 1. Participation and antigenic selection of different B cell compartments in SLE flares using repertoire analysis by NGS and single cell antibody production; Aim 2. Relative contribution of short-lived and long-lived antibody secreting cells to the SLE serum autoantibody proteome; and Aim 3. Epigenetic and transcriptional control of SLE B cells in conjunction with the Collaborative Project. The work proposed is highly synergistic with the Collaborative Agenda and the Pilot Project and has a high degree of innovation in terms of the questions asked and the experimental approach. The expect results should be of far-reaching significance for our understanding of the pathogenic mechanisms acting upon B cells in human autoimmunity and for the rationale design of safer and more effective therapies. RELEVANCE: This project will provide critical insight into the contribution of different B cells and antibody secreting cells to the autoantibody repertoire and disease process in human SLE. Together with the Collaborative Project, it will also study the epigenome and transcriptional program of autoimmune B cells.