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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要系统性硬化症（SSc）是一种多系统疾病，是主要风湿病中死亡率最高的疾病疾病。尽管某些基因位点与 SSc 易感性密切相关，但有效的开发对 SSc 的治疗因对已确定的遗传机制的支离破碎的理解而受到阻碍。变异导致 SSc。目前的提案旨在阐明一种机制，通过该机制，脱氧核糖核酸酶 1 样 3 (DNASE1L3)，导致氨基酸 206 (R206C) 处的 Arg 替换为 Cys 有助于 SSc 发病机制。 PL 的初步数据表明，患有 SSc 的患者循环基因组 DNA 的消化受损 DNASE1 L3 R206C 变体。该提案的目标 1 旨在进一步定义循环 DNA 谱存在 DNASE1 L3 R206C 的 SSc 患者。将分析循环 DNA 的两个特定群体——核小体与凋亡来源的细胞外囊泡相关的 DNA 和中性粒细胞中含有的 DNA 细胞外陷阱。 DNASE1 L3 在线粒体 DNA 消化中的潜在作用也将得到检验。下一代测序将用于确定循环 DNA 片段的长度差异。将开发并验证用于测量循环 DNASE1L3 的 ELISA。目标 2 将阐明使用树突状细胞，在 DNASE1 L3 缺乏的情况下发生的免疫细胞功能异常小鼠和人类免疫细胞的功能测定和单细胞 RNA 测序分析。一起，这些研究将揭示 DNASE1 L3 的下游生化和免疫学后果功能障碍，加强开发治疗 SSc 的靶向疗法的基础。 Pl 是一位风湿病医学博士/博士，其长期目标是成为一名专注于关于 SSc 发病机制。拟议的研究和培训计划将帮助PL培养熟练程度基因组学和转录组学数据分析以及适应性研究的实验方法免疫学。这项工作将在优秀的机构环境中进行，并得到博士的指导。 Shervin Assassi，SSc 分子分析领域的领导者，以及具有专业知识的共同导师的额外指导在生物信息学和免疫学领域。该奖项将加速 PL 的研究和职业发展成为一名独立研究员。