The Role of an MSH6 Gene Variant and UV Radiation in Systemic Lupus Erythematosus

MSH6 基因变异和紫外线辐射在系统性红斑狼疮中的作用

• 批准号: 10054266
• 负责人: Rithy Meas
• 金额: $ 8.75万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-12 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10054266
• 关键词: Affect; Affinity; Aging; Amino Acids; Antibodies; Antibody Formation; Antinuclear Antibodies; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmunity; B-Lymphocytes; Binding; Biological Assay; Cell Culture Techniques; Cell Death; Cell Survival; Cessation of life; Charge; Clone Cells; Codon Nucleotides; Collaborations; Complex; DNA; DNA Damage; DNA Repair; DNA Repair Gene; DNA lesion; Data; Dermatitis; Development; Diagnostic; Disease; Environmental Exposure; Environmental Risk Factor; Epitopes; Excision; Exposure to; Fibrinogen; Genetic; Genetic Polymorphism; Genetic Risk; Glomerulonephritis; Goals; High-Throughput Nucleotide Sequencing; Human; Immunoglobulin Class Switching; Immunoglobulin Genes; Immunoglobulin Somatic Hypermutation; Immunoglobulin Switch Recombination; Immunologics; Immunology; In Vitro; Individual; Inflammation; Knock-in; Knowledge; Lead; Link; Literature; Lupus; MSH6 gene; Mentors; Mismatch Repair; Modeling; Molecular; Mus; Other Genetics; Pathogenesis; Pathway interactions; Patients; Phase; Phenotype; Process; Production; Radiation Induced DNA Damage; Research; Role; Sequence Analysis; Serum; Severities; Single Nucleotide Polymorphism; Somatic Cell; Systemic Lupus Erythematosus; Techniques; Testing; Time; To autoantigen; Training; UV Radiation Exposure; UV induced; UV response; Ultraviolet Rays; Variant; Wild Type Mouse; Work; autoreactive B cell; autoreactivity; base; cohort; combat; ds-DNA; environmental stressor; experimental study; genetic variant; human model; in vitro Assay; insight; lupus-like; member; mouse model; novel; novel therapeutics; programs; repaired; response; student mentoring; symposium; systemic autoimmune disease; therapeutic target; tool; ultraviolet; ultraviolet irradiation

PROJECT SUMMARY/ABSTRACT Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease that involves the combination of both genetic and environmental risk factors for disease in humans. This disease afflicts over 3 million people worldwide, and there is currently no cure for SLE. Recently, I have developed a novel mouse model that has a knockin of a human SLE-associated gene variant (GV) in the mismatch repair gene MSH6. Interestingly, preliminary data indicate that mice homozygous for the MSH6 GV have significantly increased serum titers of anti-nuclear antibodies, a hallmark criterion of SLE, as compared to wildtype mice. The main objective of this study is to analyze two human-associated factors, one environmental and the other genetic, to understand the underlying mechanism of SLE pathogenesis. Importantly, this study is significant because the use of two human- associated factors, the human-derived GV and UV radiation, will closely mimic the multifactorial pathoetiology seen in human SLE. Therefore, this mouse study can potentially provide insight into human SLE that cannot be derived from other models. This proposal is divided into 3 aims to elucidate the role of UV radiation and DNA repair in SLE pathogenesis. The first aim will seek to understand if the MSH6 GV and UV radiation leads to decreased latency and/or increased severity of lupus in mice by analyzing the onset of SLE phenotypes—such as anti-nuclear antibodies, glomerulonephritis, and dermatitis—in MSH6 GV mice un/treated with UV radiation. The second aim will seek to understand the immunological effects induced by the MSH6 GV and UV radiation. This will be determined by utilizing high-throughput sequencing and in vitro assays to elucidate how the MSH6 GV and UV radiation influences antibody diversification and the removal of autoreactive B cells. Lastly, the third aim will utilize in vitro assays to determine the binding affinity of the MSH6 variant to its cognate substrate and determine if the MSH6 GV and UV radiation promote an aberrant DNA response and cell death, which can potentially induce the production of anti-nuclear antibodies by manipulating antibody diversification and autoantigen release, respectively. The first aim will be initiated and completed during the mentored phase, and the second and third aims will be initiated during the mentored phase to gain expertise in specific techniques and completed during the independent phase. To achieve the aims of this proposal, I assembled a team of mentors with expertise in human and murine autoimmunity, inflammation, immunology, and DNA repair. This team will also provide me the guidance and tools to transition into independence and to establish a successful lab. Additional opportunities to attend and present at conferences and to mentor students and postdoctorates will be complementary for my long-term goal of establishing an independent research program. Collectively, each of these individuals and opportunities will be essential for me to gain the technical training and establish a successful research program to study the role of DNA repair and environmental stressors in autoimmunity.

项目总结/摘要 系统性红斑狼疮（SLE）是一种多因素自身免疫性疾病，涉及以下因素的组合： 遗传和环境因素对人类疾病的影响。这种疾病折磨着300多万人 目前还没有治愈SLE的方法。最近，我开发了一种新颖的小鼠模型， 错配修复基因MSH 6中的人SLE相关基因变体（GV）的敲入。有趣的是， 初步数据表明，MSH 6 GV纯合子小鼠的血清滴度显著增加， 抗核抗体，SLE的标志性标准，与野生型小鼠相比。的主要目标 这项研究分析了两个与人类相关的因素，一个是环境因素，另一个是遗传因素，以了解 SLE发病的潜在机制。重要的是，这项研究是重要的，因为使用两个人类- 相关因素，人源性GV和紫外线辐射，将密切模拟多因素病因学 在人类SLE中可见。因此，这项小鼠研究可以潜在地提供对人类SLE的深入了解， 源自其他模型。本提案分为3个部分，旨在阐明紫外线辐射和DNA的作用 修复在SLE发病机制中的作用。第一个目标将试图了解MSH 6 GV和紫外线辐射是否会导致 通过分析SLE表型的发作， 作为抗核抗体，肾小球肾炎和皮炎-在MSH 6 GV小鼠中未用UV辐射处理/用UV辐射处理。 第二个目标将试图了解由MSH 6 GV和UV辐射诱导的免疫效应。 这将通过利用高通量测序和体外测定来确定，以阐明MSH 6 GV和UV辐射影响抗体多样化和自身反应性B细胞的去除。最后，第三 aim将利用体外测定来确定MSH 6变体对其同源底物的结合亲和力， 确定MSH 6 GV和UV辐射是否促进异常DNA反应和细胞死亡，这可以 通过操纵抗体多样化潜在地诱导抗核抗体的产生， 自身抗原释放。第一个目标将在辅导阶段启动和完成， 第二个和第三个目标将在指导阶段启动，以获得特定技术的专业知识 并在独立阶段完成。为了实现这一目标，我组建了一个团队， 导师在人类和小鼠自身免疫，炎症，免疫学和DNA修复的专业知识。这 团队还将为我提供指导和工具，以过渡到独立，并建立一个成功的 实验室参加和出席会议以及指导学生和博士后的额外机会 将是我建立一个独立的研究计划的长期目标的补充。总的来说， 这些人和机会中的每一个都对我获得技术培训和建立一个 成功的研究计划，以研究DNA修复和环境应激源在自身免疫中的作用。