Engineered DNA-particles to model immune events in systemic lupus erythematosus

工程 DNA 颗粒模拟系统性红斑狼疮的免疫事件

• 批准号: 10644574
• 负责人: CHRISTINE K PAYNE
• 金额: $ 23.76万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-03 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644574
• 关键词: Address; Adsorption; Affect; African American; African American population; American; Animal Model; Antibodies; Antigen-Antibody Complex; Antigens; Antinuclear Antibodies; Apoptosis; Autoimmune Diseases; Automobile Driving; Base Pairing; Behavior; Binding; Biochemical; Biological; Biological Assay; Biological Markers; Biological Models; Blood; Blood Vessels; Cell Death; Cell Line; Cell model; Cells; Classification; Coated vesicle; Complement; Complex; Confocal Microscopy; DNA; DNA Binding; Data; Deposition; Development; Diagnosis; Diameter; Digestion; Disease; Elements; Engineering; Epitopes; European; Event; Future; Immune; Immune response; Immunologic Stimulation; Immunological Models; Immunologics; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Injury; Injury to Kidney; Interleukin-6; Kidney; Length; Liquid substance; Lupus; Macrophage; Malignant - descriptor; Measures; Membrane; Molecular Weight; Monoclonal Antibodies; Morbidity - disease rate; Mus; Nephritis; Nitric Oxide; Nuclear; Nucleic Acids; Organ; Outcomes Research; Pathogenesis; Patients; Phagocytes; Polystyrenes; Production; Property; Proteins; Research; Research Personnel; Role; Serology; Source; Surface; System; Systemic Lupus Erythematosus; TNF gene; Tissues; cell free DNA; child bearing; cytokine; experimental study; extracellular; extracellular vesicles; improved; in vivo; innovation; insight; mortality; nano; nanoparticle; novel therapeutic intervention; nuclease; particle; prototype; response; serological marker; side effect; synthetic construct; tissue culture; uptake; young woman

Summary/Abstract Systemic lupus erythematosus (SLE or lupus) is a prototypic autoimmune disease that causes severe systemic manifestations, of which nephritis is the major cause of morbidity and mortality. SLE primarily affects young women, with African American patients showing much earlier and more severe disease than European Americans. An important feature of SLE is the expression of antibodies to nuclear molecules (anti-nuclear antibodies (ANAs)). ANAs provide serological markers for diagnosis, classification, and disease activity. While there has been extensive study of the pathogenesis of SLE, much remains unknown about the underlying mechanisms that promote inflammation and renal injury. As a result, SLE treatment is empiric and frequently ineffective. Current treatments can cause permanent organ damage and severe side effects, providing a strong rationale for the mechanistic studies necessary for more effective and less toxic therapies. In patients as well as animal models of SLE, the formation of immune complexes by ANAs is a key step in inflammation and injury. Of ANAs that can form immune complexes, antibodies to DNA (anti-DNA) have a prominent and well-validated role in nephritis, as shown by the isolation of anti-DNA from affected kidneys of patients, as well as the induction of nephritis in animal models by administration of monoclonal anti-DNA antibodies. While most models for immune complexes are based on antibody interaction with soluble protein or nucleic acid antigens of relatively low molecular weight, recent research suggests that immune complexes in SLE form on cell-generated particles known as extracellular vesicles. We propose that DNA can adsorb onto the surface of these particles to form a “corona” that provides a target for anti-DNA antibodies. The significance of this research is the development of well-controlled synthetic DNA-particles to address fundamental questions about the immunological properties of particles that are not addressable with naturally occurring particles. The outcome of this research will be the understanding of the mechanisms by which DNA binds to particles to form an antigenic substrate; the formation of immune complexes by particles; the effects of surface DNA on the interaction of particles with immune cells; and the role of surface DNA in immune stimulation. Aim 1 will determine the antigenicity of DNA adsorbed on particles as a function of particle diameter, DNA length, and a protein corona using a previously developed antibody binding assay as a readout. Aim 2 will use the antigenic particles identified in Aim 1 to elucidate the immunostimulatory activity of DNA-particles in vitro, measuring the immune response and cellular internalization of the DNA-particles. The development of a well-controlled synthetic particle system provides an innovative approach to the study of SLE. Beyond SLE, the role of naturally occurring, cell-generated particles as elements in disease pathogenesis has important implications for other autoimmune and inflammatory diseases. 0

总结/摘要 系统性红斑狼疮（SLE或狼疮）是一种原型自身免疫性疾病， 肾炎是引起发病和死亡的主要原因。SLE主要影响年轻人 非洲裔美国人的患者比欧洲人更早出现和更严重的疾病 美国人SLE的一个重要特征是表达针对核分子的抗体（抗核抗体）。 抗体（ANA））。ANA为诊断、分类和疾病活动提供血清学标记。而 SLE的发病机制已经有了广泛的研究，但其潜在的发病机制仍不清楚。 促进炎症和肾损伤的机制。因此，SLE治疗是经验性的， 无效。目前的治疗可能会导致永久性器官损伤和严重的副作用， 更有效、毒性更小的疗法所需的机制研究的原理。在患者中， 在SLE动物模型中，ANA形成免疫复合物是炎症和损伤的关键步骤。的 ANA能形成免疫复合物，抗DNA抗体（anti-DNA）具有突出的和充分验证的作用 在肾炎中，如从患者的受影响肾脏中分离出抗DNA所示，以及诱导 通过施用单克隆抗DNA抗体在动物模型中观察肾炎。虽然大多数免疫模型 复合物基于抗体与相对低的可溶性蛋白质或核酸抗原的相互作用， 分子量，最近的研究表明，SLE中的免疫复合物形成于细胞产生的颗粒上， 称为细胞外囊泡。我们认为DNA可以吸附在这些颗粒的表面上形成一个 “冠”为抗DNA抗体提供了靶点。本研究的意义在于 良好控制的合成DNA颗粒，以解决有关免疫特性的基本问题， 不能用天然存在的粒子寻址的粒子。这项研究的结果将是 理解DNA与颗粒结合形成抗原底物的机制; 免疫复合物的颗粒;表面DNA的颗粒与免疫细胞的相互作用的影响; 以及表面DNA在免疫刺激中的作用。目的1将确定吸附的DNA的抗原性 粒子作为粒子直径，DNA长度和蛋白质冠的函数，使用先前开发的 抗体结合测定作为读数。目标2将使用目标1中鉴定的抗原颗粒来阐明 体外DNA颗粒的免疫刺激活性，测量免疫应答和细胞内化 的DNA颗粒。良好控制的合成粒子系统的开发提供了一种创新的 SLE的研究方法。除了SLE，自然发生的细胞产生的颗粒作为元素的作用 在疾病发病机制中对其他自身免疫性和炎性疾病具有重要意义。 0