B Cell Expressed COX2 and AID Dependent Sjogrens Syndrome Autoimmunity

B 细胞表达 COX2 和 AID 依赖性干燥综合征自身免疫

基本信息

批准号：
8303999
负责人：
Patricia K. Mongini
金额：
$ 20.68万
依托单位：
FEINSTEIN INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8303999
关键词：
Acinar Cell Affinity Age Age-Years American Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Autoimmunity B lymphoid malignancy B-Cell Lymphomas B-Lymphocytes Binding Biological Assay CD22 gene Cell Lineage Cells Characteristics Cytosine deaminase DNA Development Disease Environment Enzyme Activation Enzymes Event Exocrine Glands FDA approved Fluorescence Future Generations Genes Genetic Genetic Recombination Goals Green Fluorescent Proteins Histidine Human Immunoglobulin Class Switching Immunoglobulin G In Vitro Inflammation Inflammatory Injection of therapeutic agent Laboratories Lacrimal gland structure Link Lymphoid Tissue Malignant Neoplasms Mediating Methods Monitor Mus Muscarinic Acetylcholine Receptor Mutation N-terminal Normal Cell Nuclear Oncogenic PTGS2 gene Pathology Pathway interactions Peptides Pharmaceutical Preparations Play Population Production Prostaglandin-Endoperoxide Synthase Prostaglandins Protein translocation Proteins Public Health Recombinants Reporting Research Risk Role Salivary Salivary Glands Signal Transduction Site Sjogren&apos s Syndrome Somatic Mutation Spleen Stimulus Structure of germinal center of lymph node T-Lymphocyte Terminator Codon Testing Therapeutic Therapeutic Uses Time Woman Work Xerostomia anti-IgG biodegradable polymer chronic autoimmune disease congenic cyclooxygenase 2 human disease in vivo innovation insight lymph nodes macrophage mouse model nanoparticle novel overexpression prevent promoter protein activation recombinase response saliva secretion targeted delivery therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Inflammation often promotes formation of tertiary lymphoid tissue, containing B cell foci which cause autoimmune pathology and occasionally transform into malignancy. In these sites, a DNA-modifying enzyme, activation-induced cytosine deaminase (AID), has an important role in generating pathogenic autoantibody (autoAb) and inducing oncogenic mutations. This laboratory's recent in vitro discovery that a cyclooxygenase (COX-2) axis within in vitro replicating human B cells significantly augments AID expression/function provide new insights into a mechanism for these events. This proposal's long term goal is to test the hypothesis that in vivo-expressed B cell COX-2 plays a significant role in augmenting AID and downstream pathogenic IgG autoAb in an autoimmune setting. Evidence that COX-2 activity is important for IgG Ab production in normal and autoimmune mice has been reported. Nevertheless, it remains unclear whether COX-2 function in B cells or in neighboring non-B cells is involved. Sjogren's Syndrome (SjS) is an autoimmune disease characterized by salivary gland inflammation, AID-expressing B cells, pathogenic IgG autoAbs that impair salivation, and a high rate of B cell lymphoma. Furthermore, stimuli linked to SjS in humans and/or mouse models are known to be inducers of B cell COX-2 expression in vitro. Thus, SjS is clearly a disease in which B cell COX-2 expression may be relevant. This project will test whether B cell-expressed COX-2 is important in promoting IgG Ab-driven disease manifestations in SjS. Crucial to this is (a) a recently generated SjS- susceptible NOD.B10 mouse line whose floxed COX-2 gene can be a target for Cre-mediated inactivation and (b) a proprietary method for generating mAb-bound nanoparticles (NP) containing biologically active molecules. B cell-targeted, biodegradable NP containing rCre as a "payload" will be generated with an electrohydrodynamic approach that incorporates mAb into the outer biodegradable shell and Cre into an inner core. Proof of principle studies to confirm in vitro and in vivo NP specificiy and activity will be performed with mice expressing the fluorescence indicator gene (GFP) under the control of a ubiquitous promoter and floxed stop signal. Subsequently, B cell-targeted, Cre-laden nanoparticles will be tested in SjS-susceptible NOD.B10 mice expressing floxed or wild-type COX-2 genes. Evidence that selective Cre-mediated COX-2 inactivation in B cells prevents the hyposalivation characteristic of SjS and impairs the formation of pathogenic IgG Abs to the M3R muscarinic acetylcholine receptor would be important and would suggest that specific NP hold promise in the selective targeting of therapeutic molecules in human disease. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because it will discern whether a newly discovered COX-2 pathway in B lymphocytes is important for generating pathogenic autoantibodies in autoimmune disease. While studies will be performed in a mouse model for Sjogren's Syndrome, a debilitating autoimmune disease which primarily targets women over the age of fifty, the implications of the work will extend to other conditions. Results from this project should show whether B cell COX-2 should be targeted and provide "proof of principle" evidence supporting an innovative means of targeting therapeutics selectively to B cells.

描述（由申请人提供）：炎症通常会促进三级淋巴组织的形成，其中包含引起自身免疫性病理并偶尔转化为恶性肿瘤的B细胞灶。在这些位点，激活诱导的胞嘧啶脱氨酶（AID）的DNA修饰酶在产生致病性自身抗体（AUTOAB）和诱导致癌突变方面具有重要作用。该实验室最近在体外发现，即在体外复制人B细胞内的环氧合酶（COX-2）轴显着增强辅助表达/功能，为这些事件的机制提供了新的见解。该提议的长期目标是检验以下假设：体内表达的B细胞COX-2在自身免疫环境中增强辅助和下游致病性IgG AutoAB中起着重要作用。已经报道了COX-2活性对于正常和自身免疫小鼠的IgG AB产生很重要的证据。然而，尚不清楚COX-2在B细胞中还是涉及邻近的非B细胞中的功能。 Sjogren综合征（SJS）是一种自身免疫性疾病，其特征是唾液腺炎症，表达辅助的B细胞，致病性IgG自动促进症和B细胞淋巴瘤的高率。此外，已知与人类和/或小鼠模型中SJ相关的刺激是B细胞COX-C-2在体外表达的诱导剂。因此，SJS显然是B细胞COX-2表达可能相关的疾病。该项目将测试B细胞表达的COX-2对于促进SJS中IgG AB驱动的疾病表现是否重要。至关重要的是（a）最近生成的SJS-易感性点头。B10小鼠系的floxed Cox-2基因可以成为CRE介导的失活的靶标，以及（b）一种产生包含生物学活性分子的MAB结合纳米颗粒（NP）的专有方法。 B细胞靶向的，可生物降解的NP包含RCRE作为“有效载荷”的NP将使用电水动力学方法产生，该方法将mAb掺入外部可生物降解的壳中，并将其掺入内核中。原理研究证明以确认体外和体内NP具体和活性，将使用在无处不在的启动子和Floxed Stop信号的控制下表达荧光指示基因（GFP）的小鼠进行。随后，将在表达Floxed或野生型COX-2基因的SJS敏感的点头小鼠中测试以B细胞靶向的含有CRE的纳米颗粒。 B细胞中选择性CRE介导的COX-2灭活的证据证明了SJS的造成多种特征，并且会损害致病性IgG ABS向M3R毒蕈碱乙酰胆碱受体的形成，并表明特定的NP在人类疾病中的治疗性分子的选择性靶向中会很重要。公共卫生相关性：拟议的研究与公共卫生有关，因为它会辨别出新发现的B淋巴细胞中新发现的COX-2途径对于在自身免疫性疾病中产生致病性自身抗体很重要。虽然研究将在小鼠模型中用于Sjogren综合征，这是一种使人衰弱的自身免疫性疾病，主要针对五十岁以上的女性，但该工作的含义将扩展到其他疾病。该项目的结果应显示B细胞COX-2是否应靶向，并提供“原理证明”证据，以选择性地靶向B细胞的疗法的创新手段。