2006 Electronic presentation - split rate modules

2006 电子演示 - 分流费率模块

基本信息

批准号：
8076803
负责人：
PATRICK S LEUNG
金额：
$ 18.19万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8076803
关键词：
3&apos Untranslated Regions Address Affinity Animal Model Antibodies Antigen-Presenting Cells Apolipoprotein E Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Autoimmune Responses Biliary Biological Models C57BL/6 Mouse CD80 gene CD8B1 gene Cell membrane Cell physiology Cell surface Chemicals Cholangitis Clinical Complementary DNA Data Databases Dendritic Cells Disease Electronics Elements Enhancers Epithelial Cells Extravasation Gene-Modified Goals Hepatic Hepatocyte Histopathology Human Immune Tolerance Immune response Immunization Immunobiology Immunoglobulins Immunosuppressive Agents Immunotherapy Inner mitochondrial membrane Laboratories Lead Limb structure Liver Location Luciferases Membrane MicroRNAs Mitochondria Modeling Monitor Mus Nature Orphan Patients Pharmacologic Substance Primary biliary cirrhosis Recombinants Reporter Serotyping Serum Site System T-Lymphocyte Testing Therapeutic Time Tissues Transmembrane Domain Viral Xenobiotics adeno-associated viral vector anergy combinatorial design experience gene therapy in vivo macrophage mouse model novel optical imaging overexpression prevent promoter public health relevance pyruvate dehydrogenase pyruvate dehydrogenase complex E2 response transgene expression vector vector control

项目摘要

DESCRIPTION (provided by applicant): Primary biliary cirrhosis is considered a model autoimmune disease because of the clinical homogeneity between patients and the highly specific and highly directed nature of the hallmark of this disease, sera antimitochondrial antibodies. Considerable data from our laboratory suggests that a multi-lineage response to pyruvate dehydrogenase (PDC-E2), the major mitochondrial autoantigen, is the pathogenic effector mechanism responsible for the destruction of small biliary epithelial cells (BECs). Our laboratory has considerable experience in defining the immunobiology of this immune response in humans and recently has developed a unique murine system in which a PBC-like disease is induced following chemical xenobiotic immunization. Our goal is to take advantage of these data and develop specific immunotherapy that we hypothesize will down-regulate the immune response to PDC-E2 and modify the autoimmune response. We will construct an AAV8 vector containing the ApoE/hAAT enhancer/promoter to drive PDC-E2 expression with or without four consecutive miR-155 target sites in the 3' untranslated region. Two AAV constructs will be designed. One will be confined to PDC-E2 expression on hepatocytes and the other will be a plasma membrane-anchored PDC-E2. Control vectors will similarly be produced and transduction rates and duration of transgene expression will be monitored by real time expression using an optical imaging system. Further, four perfectly complementary targets of microRNA 155 (miR-155), expression of which is found only in mature APCs, will be inserted into the 3' untranslated region of the PDC-E2 cDNA to prevent APC activation. We will apply combinatorial testing of both of these constructs in the presence or absence of miR-155 to determine whether we can inhibit anti-PDC-E2 responses in mice immunized with PDC-E2 and similarly we will determine whether specific tolerance can be reintroduced against PDC-E2 in mice with a previously established anti-PDC-E2 response. Finally, we will take advantage of our xenobiotic model of autoimmune cholangitis and the identical vectors with and without miR-155 to either prevent or modify clinical disease. In this model we can address the titer immunoglobulin subclass and affinity of the antimitochondrial response as well as examine extensive tissue histopathology and finally immunochemical characterization of liver specific infiltrates. PUBLIC HEALTH RELEVANCE: Primary biliary cirrhosis (PBC) is an orphan autoimmune disease that has been relatively ignored by the large pharmaceutical companies. There have not been recent advances in the treatment of PBC. Our goal is to take advantage of our data on the immune response in patients with PBC, our recently developed animal model of PBC and our expertise in gene therapy, to develop novel treatments that we believe will down-regulate/modulate the autoimmune response in PBC and lead to therapeutic improvement.

描述（由申请人提供）：由于患者之间的临床同质性以及该疾病标志的高度特异性和高度定向的性质，血清抗降节抗体抗体，原发性胆道肝硬化被认为是一种自身免疫性疾病。我们实验室的大量数据表明，对丙酮酸脱氢酶（PDC-E2）的多种措施是主要的线粒体自身抗原，是导致小型胆道上皮细胞（BEC）破坏的致病效应机制。我们的实验室在定义人类免疫反应的免疫生物学方面具有丰富的经验，并且最近开发了一种独特的鼠系统，在该系统中，在化学异种生物免疫后引起了PBC样疾病。我们的目标是利用这些数据并开发我们假设的特定免疫疗法，将下调对PDC-E2的免疫反应并修改自身免疫反应。我们将构建一个包含APOE/HAAT增强子/启动子的AAV8矢量，以在3'未翻译区域中使用或不连续四个连续miR-155目标位点驱动PDC-E2表达。将设计两个AAV结构。一个将仅限于肝细胞上的PDC-E2表达，另一个将是质膜锚定的PDC-E2。将同样产生控制载体，并且将使用光学成像系统实时表达来监测转基因表达的转导速率和持续时间。此外，仅在成熟的APC中发现的MicroRNA 155（miR-155）的四个完美互补靶标（miR-155）将插入PDC-E2 cDNA的3'未翻译区域中，以防止APC激活。我们将在存在或不存在miR-155的情况下对这两种构建体进行组合测试，以确定我们是否可以抑制用PDC-E2免疫的小鼠中的抗PDC-E2反应，同样，我们将确定是否可以通过先前建立的抗PDC-E2响应在小鼠中对PDC-E2进行具体耐受。最后，我们将利用自身免疫性胆管炎的异种模型和具有miR-155的相同载体，以预防或改变临床疾病。在此模型中，我们可以解决抗核糖软骨反应的滴度免疫球蛋白亚类和亲和力，并检查肝脏特异性浸润的广泛组织组织病理学和最终的免疫化学表征。公共卫生相关性：初级胆道肝硬化（PBC）是一种孤儿自身免疫性疾病，被大型制药公司相对忽略。 PBC治疗最近没有进步。我们的目标是利用我们最近开发的PBC动物模型以及我们在基因治疗方面的动物模型的PBC患者中的免疫反应数据，以开发新的治疗方法，我们认为这些治疗方法会下调/调节PBC中的自身免疫反应并导致治疗性改善。