Pathogenic effects of the DNASE1L3 R206C variant in systemic sclerosis

DNASE1L3 R206C 变异在系统性硬化症中的致病作用

• 批准号: 10685267
• 负责人: Brian Skaug
• 金额: $ 16.42万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-10 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685267
• 关键词: Acceleration; Amino Acids; Antigen-Presenting Cells; Apoptosis; Autoantibodies; Autoimmune Diseases; Award; Biochemical; Bioinformatics; Biological Assay; CREST Syndrome; Cell physiology; Cell secretion; Cells; Circulation; DNA; Data; Dendritic Cells; Deoxyribonucleases; Development; Digestion; Disease; Doctor of Philosophy; Environment; Enzyme-Linked Immunosorbent Assay; Foundations; Functional disorder; Gene Expression; Gene Expression Profiling; Genetic Polymorphism; Genomic DNA; Genomics; Goals; Helper-Inducer T-Lymphocyte; Human; Immune; Immunologics; Immunology; Impairment; Institution; Interferon Type I; Interferons; Knockout Mice; Label; Laboratories; Lead; Length; Link; Lupus; Measurement; Mentors; Mentorship; Mitochondrial DNA; Molecular; Molecular Profiling; Mus; Nucleosomes; Pathogenesis; Pathogenicity; Patients; Peripheral Blood Mononuclear Cell; Population; Research; Research Personnel; Rheumatism; Risk Factors; Role; Sampling; Susceptibility Gene; System; Systemic Scleroderma; Training; Translational Research; Variant; Work; biobank; career; career development; circulating DNA; design; effective therapy; extracellular; extracellular vesicles; genetic variant; genomic locus; improved; monocyte; mortality; mouse model; neutrophil; next generation sequencing; novel; rheumatologist; single-cell RNA sequencing; targeted treatment; transcriptomics

PROJECT SUMMARY/ABSTRACT Systemic sclerosis (SSc) is a multi-system disease with the highest mortality among major rheumatologic diseases. Though certain genetic loci have been robustly linked to SSc susceptility, development of effective therapies for SSc is hampered by a fragmented understanding of mechanisms by which the identified genetic variants lead to SSc. The current proposal is designed to elucidate mechanisms by which a variant in deoxyribonuclease 1-like 3 (DNASE1L3) that results in Arg to Cys substitution at amino acid 206 (R206C) contributes to SSc pathogenesis. The Pl's preliminary data indicate that digestion of circulating genomic DNA is impaired in SSc patients with the DNASE1 L3 R206C variant. Aim 1 of the proposal is designed to further define the circulating DNA profile in SSc patients with DNASE1 L3 R206C. Two specific populations of circulating DNA will be analyzed--nucleosomal DNA associated with apoptosis-derived extracellular vesicles and DNA contained in neutrophil extracellular traps. A potential role of DNASE1 L3 in digestion of mitochondrial DNA will also be examined. Next-generation sequencing will be employed to determine length differences in circulating DNA fragments. An ELISA for measurement of circulating DNASE1L3 will be developed and validated. Aim 2 will elucidate aberrancies in immune cell function that occur in the setting of DNASE1 L3 deficiency, using a dendritic cell functional assay and single cell RNA Sequencing analyses of murine and human immune cells. Together, these studies will shed light on the downstream biochemical and immunologic consequences of DNASE1 L3 dysfunction, strengthening the foundation for development of targeted therapies to treat SSc. The Pl is an MD/PhD Rheumatologist with the long-term goal of becoming an independent investigator focused on SSc pathogenesis. The proposed research and training plan will help the Pl to develop proficiency in analyses of genomic and transcriptomics data and in experimental approaches for the study of adaptive immunology. The work will be performed in an excellent institutional environment with mentorship from Dr. Shervin Assassi, a leader in molecular profiling of SSc, and additional guidance from co-mentors with expertise in bioinformatics and immunology. This award will accelerate the Pl's research as well as career development into an independent researcher.

项目总结/文摘