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Cytoplasmic mitochondrial dsRNA in pediatric Sjogren's syndrome

小儿干燥综合征中的细胞质线粒体 dsRNA

基本信息

批准号：
10287866
负责人：
SEUNGHEE CHA
金额：
$ 36.91万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-22 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10287866
关键词：
Address Adolescent Adult Affect Applications Grants Autoimmune Autoimmunity B-Cell Lymphomas Bacterial Infections Biological Biological Assay Biopsy Specimen CD14 gene Cell Differentiation process Cell Line Cell physiology Cells Child Childhood Clinical Research Coculture Techniques Collaborations Communication Complement Control Groups Cytoplasm Data Diagnosis Diagnostic Disease Double-Stranded RNA Elderly Female Gene Expression Gene Targeting Genes Gland Goals High Prevalence Immune Impairment Inflammation Institutes Interdisciplinary Study Interferons Interleukin-17 Investigation Knowledge Lacrimal gland structure Lead Lymphoma Mitochondria Mitochondrial RNA Mus Nature Nucleic Acids Pain Parotitis Particulate Pathogenesis Pathology Pathway interactions Patients Pediatric cohort Peripheral Peripheral Blood Mononuclear Cell Polyribonucleotide Nucleotidyltransferase Production Prognosis Property RNA Degradation RNA Helicase Recurrence Regulation Regulatory T-Lymphocyte Reporting Role Salivary Glands Signal Transduction Sjogren&apos s Syndrome Stains Swelling Tissues Transfection Transfer RNA United States National Institutes of Health Viral Virus Diseases autoimmune inflammation cohort effector T cell evidence base experimental study genetic signature helicase high risk in vitro Assay indexing innovation intercellular communication knock-down macrophage mitochondrial genome monocyte multidisciplinary novel novel therapeutics sensor single-cell RNA sequencing small hairpin RNA therapeutic target transcriptome sequencing transcriptomics

项目摘要

ABSTRACT Through our multidisciplinary clinical and research collaboration, we have recently discovered a rare cohort of children and adolescents who fulfill the Sjögren’s syndrome (SS) diagnostic criteria (pediatric SS, PSS). As autoimmune SS typically affects elderly females, whether PSS is a unique disease entity or a mere reflection of an early stage of SS is completely unknown. PSS is commonly misdiagnosed as infectious recurrent parotitis (RP), since glandular swelling and pain without a viral/bacterial infection occurs more frequently in PSS than in SS. The overall goal of this application is to characterize the pathology of robust target tissue inflammation in PSS with RP. Our recent, preliminary discovery by RNA-sequencing of SS monocytes revealed aberrant expression of mitochondrial-dsRNA (mtdsRNA) regulators such as SUV3 (ATP-dependent RNA helicase, suppressor of variegation 3), along with the type I IFN signature and a tendency of M1 polarization-associated genes. Consistently, SUV3 gene expression was downregulated in PSS monocytes, detected by RT-qPCR, and knockdown of SUV3 by shRNA transfection led to massive accumulation of mtdsRNA in a cell line. More interestingly, cytoplasmic accumulation of particulate dsRNA was detected in PSS and SS monocytes, which prompted us to ask fundamental questions regarding the nature of the detected dsRNA and its potential impact on monocytes and subsequent immune dysregulation in PSS. To address these questions, we propose our central hypothesis that aberrant mtdsRNA degradation by altered RNA helicase expression and activity elicits amplified priming sensitivity of PSS monocytes by mimicking a viral infection. Our two Specific Aims are: Aim 1. To characterize the nature of cytoplasmic dsRNA involving SUV3 in monocytes of PSS. We hypothesize that cytoplasmic accumulation of mtdsRNA by aberrant mtdsRNA degradation triggers type I IFN production in monocytes of PSS. Cytoplasmic dsRNA transfer and mitochondrial genome depletion will be applied to identify potential mitochondriopathies involving SUV3, which will be complemented by RNA-sequencing of purified monocytes. Aim2. To determine the impact of dsRNA- positive monocytes on macrophage polarization and regulatory T cell (Treg) conversion. We hypothesize that dsRNA and type I IFN signature in PSS monocytes confer propensity toward intensified M1polarization impacting T reg plasticity. Various in vitro assays, such as co-culture experiments and functional assays, will be carried out in parallel with scRNA-seq of PMBC and paired biopsy specimens to profile cellular communications between macrophage and Tregs in the target tissue, which we hypothesize to be influenced by cytoplasmic dsRNA in PSS monocytes. To our knowledge, this is the first study to delineate the properties and impact of cytoplasmic mtdsRNA on monocyte dysregulation and robust inflammation in RP, as well as the first investigation of PSS pathogenesis in the field. Our novel hypothesis involving mtRNA helicases will provide a new opportunity to fill the current knowledge gap.

摘要