Project 3: The role of DNASE1L3 and its DNA substrate in lupus

项目3：DNASE1L3及其DNA底物在狼疮中的作用

基本信息

批准号：
10004507
负责人：
Boris Reizis
金额：
$ 28.66万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-22 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10004507
关键词：
Affinity Animal Model Antibodies Antibody Formation Antigen-Antibody Complex Antigens Apoptotic Autoantibodies Autoantigens B cell repertoire Benign Blood Circulation Cells Chromatin Clinical DNA Data Deoxyribonucleases Disease Flare Genomic DNA Histones Human Immune Complex Glomerulonephritis Immunoglobulin G In Vitro Incidence Inflammation Longitudinal Studies Lupus Membrane Modeling Molecular Molecular Profiling Mus Nature Nuclear Antigens Nucleosomes Outcome Pathogenicity Patients Ploidies Prevention therapy Proteins Proteomics Reporting Research Personnel Ribonucleoproteins Role Sampling Severities Surface Systemic Lupus Erythematosus Technology Testing Therapeutic Therapeutic Agents Therapeutic Uses Tissues Variant autoreactivity base clinically relevant ds-DNA early onset immune activation in vivo in vivo evaluation insight novel strategies null mutation pre-clinical prevent protein complex response serological marker therapeutic evaluation

项目摘要

ABSTRACT The hallmark of systemic lupus erythematosus (SLE) is the production of antibodies to nuclear antigens such as ribonucleoproteins and DNA. Antibodies to double-stranded DNA (dsDNA) and/or chromatin represent an important transition from benign to overt clinical SLE and may predict flares and the severity of tissue damage. The mechanisms of tolerance to chromatin/DNA and of its breakdown in SLE are poorly understood. We explored these mechanisms by focusing on DNASE1L3, a unique secreted DNase whose null mutations are associated with an early-onset familial SLE. Our studies showed that chromatin in microparticles derived from apoptotic cells represented a specific substrate of DNASE1L3 in vitro and in vivo. They also demonstrated that the chromatin and/or other antigens were exposed on the surface of microparticles and recognized by autoantibodies in a DNASE1L3-sensitive manner. We hypothesize that DNASE1L3-sensitive DNA/protein complexes on microparticles are important self-antigens in human SLE, and that that DNASE1L3 can be used to target them for therapeutic purposes. The collaborative nature of the project and unique patient samples and technologies available from our co-investigators will be leveraged to develop and test this hypothesis. In Aim 1, the incidence and clinical relevance of antibodies to DNASE1L3-sensitive antigens in SLE patients will be explored. In Aim 2, DNASE1L3-sensitive antigens on microparticles and the antibodies targeting them will be characterized at the molecular level. In Aim 3, the utility of DNASE1L3 as a therapeutic agent will be tested in animal models of SLE. Collectively, these studies would provide insights into the origin and mechanisms of the pathogenic responses to DNA and associated antigens in human SLE, and facilitate novel approaches towards their therapeutic blockade.

抽象的系统性红斑狼疮 (SLE) 的标志是产生针对核抗原的抗体，例如如核糖核蛋白和DNA。双链 DNA (dsDNA) 和/或染色质抗体代表从良性到明显临床 SLE 的重要转变，可以预测耀斑和组织损伤的严重程度。 SLE 中染色质/DNA 的耐受机制及其分解机制尚不清楚。我们通过关注 DNASE1L3 来探索这些机制，DNASE1L3 是一种独特的分泌性 DNase，其无效突变是与早发的家族性 SLE 相关。我们的研究表明，微粒中的染色质来源于凋亡细胞代表 DNASE1L3 在体外和体内的特定底物。他们还证明了染色质和/或其他抗原暴露在微粒表面并被以 DNASE1L3 敏感方式产生自身抗体。我们假设DNASE1L3敏感的DNA/蛋白质微粒上的复合物是人类 SLE 中重要的自身抗原，并且可以使用 DNASE1L3 以达到治疗目的。该项目的协作性质和独特的患者样本以及我们的联合研究人员提供的技术将被用来开发和测试这一假设。瞄准 1、SLE患者中DNASE1L3敏感抗原抗体的发生率和临床相关性探索过。在目标 2 中，微粒上的 DNASE1L3 敏感抗原和针对它们的抗体将被在分子水平上进行表征。在目标 3 中，DNASE1L3 作为治疗剂的效用将在 SLE 动物模型。总的来说，这些研究将为了解疾病的起源和机制提供见解。人类 SLE 中对 DNA 和相关抗原的致病反应，并促进新的方法他们的治疗封锁。