Single-cell sequencing of peripheral blood cells in SLE patients

SLE 患者外周血细胞的单细胞测序

• 批准号: 9372979
• 负责人: Chun Jimmie Ye
• 金额: $ 19.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-14 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9372979
• 关键词: Affect; Alleles; Antinuclear Antibodies; Autoimmune Diseases; B-Lymphocytes; Behavior; Biological Assay; Biological Markers; Blood Cells; CD4 Positive T Lymphocytes; Cell physiology; Cells; Collaborations; Collection; Computational algorithm; Data; Data Set; Dendritic Cells; Disease; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Determinism; Genetic Transcription; Genotype; Immune; Immune response; Immunologics; Individual; Inflammation; Interferon-beta; Interferons; Lead; Light; Lupus; Lupus Nephritis; Mediating; Methods; Microfluidics; Molecular; Monitor; Nephritis; Noise; Outcome; Pathogenicity; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Regulator Genes; Role; Running; Sampling; Science; Severities; Severity of illness; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Systemic Lupus Erythematosus; T-Lymphocyte; Technology; Time; Transcriptional Regulation; Variant; base; cell type; cost; design; disease phenotype; experimental study; genetic information; genetic variant; genome wide association study; genome-wide; innovation; instrument; molecular pathology; nanolitre; next generation sequencing; novel strategies; pleiotropism; programs; response; single cell sequencing; therapeutic development; transcription factor; transcriptome sequencing; transcriptomics; treatment response

Systemic lupus erythematosus (SLE) has long been characterized by the prototypical expression of type-1 interferon induced genes (IFIGs). However, because of the pleiotropic roles of many IFIGs, how different peripheral blood mononuclear cells (PBMCs) mediate type-1 interferon signaling to cause disease is largely unknown. Here, we propose to use droplet-based RNA-sequencing to study the interferon response of PBMCs from lupus patients and healthy controls. Leveraging naturally occurring “genetic barcodes”, we will develop a highly innovative multiplexing strategy that significantly increases the throughput, reduces the cost, and limits unwanted technical noise of current droplet-based RNA-sequencing technologies. We will develop computational algorithms for assigning individual cells to the donor of origin and removing unwanted droplets containing multiple cells. We will use the multiplexing strategy to generate a rich dataset (~250K total cells) that enables, for the first time, the unbiased characterization of the effects of interferon-beta on PBMCs without sorting. We will first compare PBMCs from 8 healthy controls, 8 lupus and 8 lupus nephritis patients, to identify cell-type-specific interferon-beta response signatures that is predictive of disease status and severity. These signatures could be used to better monitor lupus progression and treatment response. By profiling PBMCs from 64 genotyped lupus patients, we will then characterize how common genetic variants affect cell-type-specific responses to interferon-beta, including expression parameters (e.g. variance across single cells) impossible to obtain from bulk RNA-sequencing. These results could be compared to genetic variants associated with lupus identified by genome-wide association studies to better understand the molecular pathology of the disease.

系统性红斑狼疮（SLE）一直以1型的原型表达为特征