Integrative single cell and spatial transcriptomics of salivary glands in Sjogren's syndrome

干燥综合征唾液腺的综合单细胞和空间转录组学

基本信息

批准号：
10189553
负责人：
A Darise Farris
金额：
$ 13.47万
依托单位：
OKLAHOMA MEDICAL RESEARCH FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10189553
关键词：
Address Affect Age American Antigen Presentation Apoptosis Autoantibodies Autoimmune Autoimmune Diseases B-Cell Activation B-Cell Development B-Lymphocytes Bar Codes Cell Communication Cell Migration Pathway Cell Nucleus Cell physiology Cells Clinic Clinical Clinical Data Complementary DNA Complex Couples Data Data Set Development Disease Dryness Duct (organ) structure Ductal Epithelial Cell Epithelial Cells Etiology Evaluation Exocrine Glands Exocrine System Eye Fatigue Functional disorder Gender Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Genetic Risk Genetic Transcription Genomics Genotype Gland Glass HLA Antigens Haplotypes Hematologic Neoplasms Hematoxylin and Eosin Staining Method Heterozygote IL12A gene Immune Immune system Immunity Individual Infiltration Inflammation Mediators Inflammatory Infrastructure Interferon Type II Interferon-beta Interferons Interleukin-12 Interleukins Knowledge Lacrimal gland structure Lead Lesion Life Ligase Light Link Lymphoma Malignant Neoplasms Maps Measurement Mediating Mediator of activation protein Methods Molecular Morphology Myoepithelial NF-kappa B Neurologic Neuropathy Oklahoma Oral cavity Organ PRDM1 gene Pathogenesis Pathogenicity Pathology Pathway interactions Patients Pattern Phenotype Phosphotransferases Plasma Cells Play Process Production Publishing Quality of life Quantitative Trait Loci RNA Sequences Race Research Risk Role STAT4 gene Salivary Gland Tissue Salivary Glands Sampling Signal Transduction Sjogren&apos s Syndrome Slide Small Nuclear RNA Specificity Stains Structure Structure of germinal center of lymph node Systemic disease T-Lymphocyte Therapeutic Intervention Tissues Transcript Transcriptional Regulation Translational Research Tumor-infiltrating immune cells Viral Work base cell type chemokine clinical Diagnosis clinically significant cytokine disability effective therapy falls genetic association genetic variant genomic locus insight multidisciplinary new technology new therapeutic target novel oligoadenylate preservation prevent repository response risk variant single-cell RNA sequencing targeted treatment therapeutic candidate therapeutic target therapy development transcriptome sequencing transcriptomics

项目摘要

ABSTRACT Sjögren’s syndrome (SS) is a complex autoimmune disorder that affects 2-3.1 million Americans and is distinguished by irreversible exocrine gland damage that often results in debilitating dryness in the mouth and eyes. Severe systemic manifestations may include neuropathies, lymphomas, and clinically significant fatigue leading to considerable disability and reduced quality of life. Although dysregulation of innate and adaptive immune cells undoubtedly plays a role in exocrine gland dysfunction, why salivary gland ductal cells are targeted and immune cell infiltrates form foci around these structures is poorly understood. Known genetic risk variants in SS have been mapped to loci involved in antigen presentation, interferon responses, T and B cell activation, cell migration pathways, and autoantibody production, however the cell types in which functional effects of these genes operate in SS are unknown. Our central hypothesis is that salivary gland epithelial cells contribute directly to aberrant focal immune cell infiltration through dysregulation of normal homeostatic pathways and that these interactions are driven by recently identified SS genetic risk variants. This project seeks to define gene expression patterns that differ between ductal cells with presence or absence of immune cell foci and determine if these differences vary between risk and non-risk SS genotypes. We will apply a recently described integrative approach that couples 10X Genomics-based single cell RNA-sequencing with Spatial Transcriptomics data, thus allowing cellular expression levels to be interpreted in the context of very precise tissue morphology. In Aim 1, we will define transcriptional signatures of ductal cells involved in SS by comparing gene expression patterns of ductal cells surrounded by immune foci with those that are not through intrasubject analyses of 25 SS patients. Baseline ductal cell expression patterns obtained for 25 age, gender and racially matched healthy controls will also be compared. In Aim 2, we will define specific salivary gland and immune cell subsets that are functionally affected by known SS-associated expression quantitative trait loci (eQTLs). Using the same datasets generated for Aim 1, transcript levels will be compared between carriers of risk and non-risk variants in eQTLs for genes previously associated with SS. These data will identify the specific salivary gland (e.g. ductal, acinar, myoepithelial) and immune cells (e.g. T, B, dendritic, natural killer, etc.) subsets that are functionally affected by SS risk variants. We will leverage the extensive infrastructure and multidisciplinary expertise developed through the Oklahoma Sjögren’s Syndrome Center of Research Translation to accomplish the successful execution of this project. These studies will provide unprecedented depth and breadth of new knowledge regarding cell- specific dysregulation of genes and pathways. This knowledge is essential for development of novel therapeutics that target the right genes, pathways and cells and block the interactions between the immune system and exocrine glands that are fundamental to the pathogenesis of this complex disease.

抽象的干燥综合征 (SS) 是一种复杂的自身免疫性疾病，影响 2-310 万美国人，其特点是不可逆的外分泌腺损伤，通常会导致口腔干燥和虚弱严重的全身症状可能包括神经病、淋巴瘤和临床上明显的疲劳。尽管先天和适应性失调，但仍导致严重残疾和生活质量下降。免疫细胞无疑在外分泌腺功能障碍中发挥着作用，为什么唾液腺导管细胞成为目标对于免疫细胞从这些结构周围的病灶浸润的情况知之甚少。 SS 中的位点已被定位到涉及抗原呈递、干扰素反应、T 和 B 细胞激活的位点，细胞迁移途径和自身抗体的产生，然而这些功能影响的细胞类型 SS 中的基因尚不清楚，我们的中心假设是唾液腺上皮细胞直接起作用。通过正常稳态途径失调导致异常局灶性免疫细胞浸润，并且这些相互作用是由最近发现的 SS 遗传风险变异驱动的，该项目旨在定义基因。存在或不存在免疫细胞灶的导管细胞之间的表达模式不同，并确定如果这些差异在风险和非风险 SS 基因型之间有所不同，我们将应用最近描述的综合方法。将基于 10X 基因组学的单细胞 RNA 测序与空间转录组学数据相结合的方法，从而允许在非常精确的组织形态的背景下解释细胞表达水平。在目标 1 中，我们将通过比较 SS 的基因表达模式来定义参与 SS 的导管细胞的转录特征通过对 25 名 SS 患者进行受试者内分析，发现导管细胞被免疫灶包围，而那些未被免疫灶包围的导管细胞则未被免疫灶包围。为 25 名年龄、性别和种族匹配的健康对照获得的基线导管细胞表达模式将在目标 2 中，我们将定义功能上的特定唾液腺和免疫细胞亚群。受已知 SS 相关表达数量性状位点 (eQTL) 的影响，使用生成的相同数据集。对于目标 1，将比较基因 eQTL 中风险变异和非风险变异携带者之间的转录水平以前与 SS 相关的这些数据将识别特定的唾液腺（例如导管、腺泡、肌上皮细胞）和免疫细胞（例如 T、B、树突状细胞、自然杀伤细胞等）亚群，其功能受到影响我们将利用通过开发的广泛的基础设施和多学科专业知识。俄克拉荷马州干燥综合症研究翻译中心成功执行这些研究将提供有关细胞的前所未有的深度和广度的新知识。这些知识对于开发新疗法至关重要。靶向正确的基因、途径和细胞，并阻断免疫系统和免疫系统之间的相互作用外分泌腺是这种复杂疾病发病机制的基础。