The Retroelement LINE1 and the DNA Sensor IF16 in Sjogren's Syndrome

干燥综合症中的 Retroelement LINE1 和 DNA 传感器 IF16

• 批准号: 10361213
• 负责人: Brendan Antiochos
• 金额: $ 15.88万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10361213
• 关键词: Affect; Antibodies; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmunity; Autophagocytosis; Award; Binding; Biochemical; Biological Markers; Blood; Burn injury; Cell Line; Cells; Cellular Assay; Characteristics; Complex; Cultured Cells; Cytoplasm; DNA; Data; Development; Disease; Ductal Epithelial Cell; Epithelial Cells; Filament; Fostering; Future; Generations; Genetic Materials; Gland; Grant; Human; Hybrids; Immune; Immune response; Immune system; Immunologic Surveillance; In Vitro; Inflammatory; Innate Immune Response; Interferons; Knowledge; Link; Measures; Mediating; Membrane; Mentorship; Natural Immunity; Nucleic Acids; Organ; Organelles; Pathogenesis; Pathway interactions; Patients; Persons; Physicians; Play; Process; Proteins; RNA; Research; Research Personnel; Retroelements; Retrotransposition; Reverse Transcription; Rheumatism; Role; Salivary; Salivary Glands; Salivary duct structure; Scientist; Signal Transduction; Site; Sjogren' s Syndrome; Source; Syndrome; System; Testing; Therapeutic; Tissues; Training; Visualization; Work; base; ds-DNA; genetic element; human tissue; immune activation; mouse model; multidisciplinary; new therapeutic target; patient biomarkers; patient subsets; protein expression; response; sensor; skills; symposium; therapeutic target

Project Summary/Abstract This award will promote the development of the applicant who seeks to acquire the skills and knowledge necessary to become an independent physician scientist focused on human rheumatic diseases. The research focus of this grant is the retroelement LINE1, and its potential interaction with the cytoplasmic DNA sensor IFI16, in the context of the autoimmune disease Sjögren’s syndrome (SS). Persistent activation of immune responses against nucleic acid-containing autoantigens is a hallmark of several rheumatic diseases, including SS, where an “interferon (IFN) signature” is found in the blood and affected tissues. Activation of innate nucleic acid sensors is a key driver of IFN signaling that has been implicated in SS and related diseases. In this proposal, we will examine the retroelement LINE1 as a potential endogenous driver of both the innate and humoral immune response in SS, and examine the role that autophagy plays in regulating the IFN response to LINE1. These studies will build upon our recent observation that the DNA sensor and SS autoantigen IFI16 is activated in a filamentous form in some SS salivary ductal epithelial cells – a process which seems linked to the presence of anti-IFI16 antibodies in SS sera. Preliminary data reveal that the LINE1 protein ORF1p appears to be an SS autoantigen as well, and is strikingly associated with anti-IFI16 antibodies in these patients. Moreover, LINE1 protein expression can be detected in the same ductal epithelial cell layer that harbors activated IFI16 in SS glands, suggesting that LINE1 expression in these cells could be a potential source of nucleic acid responsible for IFI16 filament formation observed at that site. Based on these preliminary findings, we hypothesize that IFI16 binding to LINE1 nucleic acids in the cytoplasm of epithelial cells could explain the association between anti- IFI16 and anti-ORF1p antibodies in some SS patients. Using cultured cells, we have also visualized co- localization of IFI16-DNA complexes with autophagy proteins, and have observed evidence of autophagy activation following cytoplasmic DNA sensing. We propose that autophagy mediates homeostatic disposal of interferogenic IFI16-LINE1 complexes, and could therefore represent an augmentable pathway in this diseases. These hypotheses will be tested using a combination of human tissues, cellular assays, and in vitro studies. This work will benefit from the expertise of two key collaborators: Dr. Kathy Burns, an expert in retroelements, and Dr. Jungsan Sohn, a biophysicist expert in innate immune sensors. In addition to completion of the proposed experimental work, the applicant will perform formal coursework, receive hands on technical training, attend relevant conferences, and obtain careful mentorship from a multi-disciplinary team of accomplished investigators over the duration of this award. Completion of this work will foster the development of the candidate into an independent physician researcher with expertise in the mechanistic study of human rheumatic disease, with particular focus on the relationship between retroelements and innate immunity.

项目摘要/摘要 该奖项将促进寻求获得技能和知识的申请人的发展 成为一名专注于人类风湿病的独立物理科学家所必需的。研究 该赠款的焦点是Retroement Line1，其与细胞质DNA传感器IFI16的潜在相互作用， 在自身免疫性疾病的背景下，Sjögren综合征（SS）。免疫复杂的持续激活 反对含核酸的自身抗原是几种风湿性疾病的标志，包括SS 在血液和受影响的组织中发现了“干扰素（IFN）特征”。先天核酸传感器的激活 是SS和相关疾病中隐含的IFN信号的关键驱动力。在此提案中，我们将 检查retroement Line1是先天和体液免疫的潜在内源驱动器 SS中的响应，并检查自噬在调节IFN对LINE1响应中的作用。这些 研究将基于我们最近的观察结果，即DNA传感器和SS自动抗原IFI16在A中激活 某些SS唾液导管上皮细胞中的丝状形式 - 似乎与存在有关的过程 SS血清中的抗IFI16抗体。初步数据表明，Line1蛋白ORF1P似乎是SS 自动抗原也与这些患者的抗IFI16抗体相关。此外，Line1 可以在同一导管上皮细胞层中检测到蛋白质表达 腺体，表明这些细胞中的线1表达可能是核酸负责的潜在来源 对于在该地点观察到的IFI16细丝形成。基于这些初步发现，我们假设IFI16 在上皮细胞的细胞质中与线1的结合可以解释抗 - 一些SS患者的IFI16和抗ORF1P抗体。使用培养的细胞，我们还可以看到 IFI16-DNA复合物与自噬蛋白的定位，并观察到自噬的证据 细胞质DNA敏感性后激活。我们建议自噬介导 干涉性IFI16-line1复合物，因此可以代表该疾病中可增强的途径。 这些假设将使用人体组织，细胞测定和体外研究的组合进行测试。这 工作将从两个主要合作者的专业知识中受益：凯西·伯恩斯博士，是重新元素专家，以及 Jungsan Sohn博士是先天免疫传感器的生物物理学专家。除了完成拟议的 实验性工作，申请人将进行正式课程，接受技术培训，参加 相关会议，并从成熟的调查人员组成的多学科团队中获得仔细的精神训练 在此奖项期间。这项工作的完成将促进候选人的发展 独立的物理研究人员具有人性风湿病机理研究的专业知识， 特别关注追溯元素与先天免疫之间的关系。