Investigating immunophenotypic and transcriptional heterogeneity as biomarkers of pain centralization in rheumatoid arthritis

研究免疫表型和转录异质性作为类风湿性关节炎疼痛集中的生物标志物

• 批准号: 10569603
• 负责人: Yvonne Claire Lee
• 金额: $ 21.12万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10569603
• 关键词: Address; Affect; Analgesics; Area; B-Lymphocytes; Biological Markers; Biometry; Blood specimen; Brain; Cells; Central Nervous System; Characteristics; Circulation; Clinical; Clinical Trials; Consumption; Data; Dendritic Cells; Development; Disease-Modifying Second-Line Drugs; Distant; Enrollment; Excision; Flow Cytometry; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genomic approach; Goals; Heterogeneity; High Prevalence; Immune; Immunophenotyping; Individual; Inflammatory; Intractable Pain; Literature; Maintenance; Measurement; Measures; Modeling; Neuronal Plasticity; Opioid; Opioid Analgesics; Outcome; Pain; Pain Measurement; Pain Threshold; Pain intensity; Pathway interactions; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Pharmacotherapy; Phenotype; Physicians; Prevention; Procedures; Process; Public Health; Regulation; Reporting; Reproducibility; Research; Resolution; Resort; Rheumatism; Rheumatoid Arthritis; Role; Sensory; Spinal Cord; Spondylarthropathies; Stimulus; Systemic Lupus Erythematosus; T-Lymphocyte; Testing; Time; Training; addiction; biomarker development; biomarker validation; cell type; central pain; chronic pain; chronic pain management; experience; functional genomics; high reward; high risk; insight; joint inflammation; monocyte; non-opioid analgesic; novel; pain processing; pain sensitivity; patient subsets; peripheral blood; precision medicine; pressure; response; single-cell RNA sequencing; success; systemic inflammatory response; trapezius muscle; Address; Affect; Analgesics; Area; B-Lymphocytes; Biological Markers; Biometry; Blood specimen; Brain; Cells; Central Nervous System; Characteristics; Circulation; Clinical; Clinical Trials; Consumption; Data; Dendritic Cells; Development; Disease-Modifying Second-Line Drugs; Distant; Enrollment; Excision; Flow Cytometry; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genomic approach; Goals; Heterogeneity; High Prevalence; Immune; Immunophenotyping; Individual; Inflammatory; Intractable Pain; Literature; Maintenance; Measurement; Measures; Modeling; Neuronal Plasticity; Opioid; Opioid Analgesics; Outcome; Pain; Pain Measurement; Pain Threshold; Pain intensity; Pathway interactions; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Pharmacotherapy; Phenotype; Physicians; Prevention; Procedures; Process; Public Health; Regulation; Reporting; Reproducibility; Research; Resolution; Resort; Rheumatism; Rheumatoid Arthritis; Role; Sensory; Spinal Cord; Spondylarthropathies; Stimulus; Systemic Lupus Erythematosus; T-Lymphocyte; Testing; Time; Training; addiction; biomarker development; biomarker validation; cell type; central pain; chronic pain; chronic pain management; experience; functional genomics; high reward; high risk; insight; joint inflammation; monocyte; non-opioid analgesic; novel; pain processing; pain sensitivity; patient subsets; peripheral blood; precision medicine; pressure; response; single-cell RNA sequencing; success; systemic inflammatory response; trapezius muscle

Over half of patients with rheumatoid arthritis (RA) report pain despite treatment with strong disease-modifying antirheumatic drugs (DMARDs). Our research group was one of the first to show that this pain is associated with increased pain sensitivity in areas distant from inflamed joints, suggestive of abnormalities in the way the brain and spinal cord regulate pain. The pain associated with this process is termed “centralized pain” and is frequently treated with opioid analgesics. However, opioids are minimally effective and often misused. The development of non-opioid analgesics has been hindered by our limited understanding of how the centralized pain phenotype relates to changes at the cellular level. In the research setting, centralized pain is often assessed using quantitative sensory testing (QST). However, QST is time-consuming, requires significant assessor training, and is prone to measurement error. There is a critical unmet need to develop quantifiable measurements of the altered cellular state that distinguish patients with centralized pain. It is imperative to address this need to achieve our long-term goal of developing safe and efficacious non-opioid pain analgesics for patients with rheumatic diseases. The objective of this high-risk, high-reward proposal is to generate cutting-edge insights into the relationship between peripheral blood mononuclear cells (PBMCs) and centralized pain in patients with RA. Our proposal emphasizes an unbiased, high-throughput approach, using multi-parameter flow cytometry and single-cell RNA sequencing (scRNA-seq) to identify characteristics of PBMCs that are associated with centralized pain after DMARD therapy. Our central hypothesis is that differences in the composition and gene expression of circulating immune cells, particularly monocytes, will characterize RA patients with a centralized pain phenotype vs. those without a centralized pain phenotype. In Aim 1, we will determine the association between the immunophenotypic profile of PBMCs and centralized pain. We will enroll 50 RA patients with minimal joint inflammation but varying levels of pain after DMARD treatment. These patients will undergo QST to test whether centralized pain, assessed by pressure pain thresholds at the trapezius muscle, is associated with: a) the proportion of each cell type in circulation, and b) continuous measures of activation status for each cell type. In Aim 2, we will identify differences in the transcriptional heterogeneity of circulating PBMCs between patients with a centralized pain phenotype vs. those without a centralized pain phenotype. We will perform scRNA-seq on a subset of patients from Aim 1 with the highest (N = 10) and lowest (N = 10) levels of centralized pain. We will investigate differences in transcriptional subpopulations and cell-type-specific gene expression between the two groups. Our proposal has high impact potential because, if successful, it will deliver novel insights into the role of circulating PBMCs in altered pain regulation in patients with RA, the prototypical systemic inflammatory condition. Data from this proposal will be used to inform an R01 application to identify a reproducible peripheral blood biomarker for centralized pain in patients with RA.

超过一半的类风湿关节炎（RA）报告了疼痛目的地治疗，并进行较强的疾病调整 抗疾病药物（DMARDS）。我们的研究小组是第一个表明这种痛苦与 增加了远离发炎关节的区域的疼痛敏感性，表明大脑的方式异常 脊髓调节疼痛。与此过程相关的疼痛称为“集中式疼痛”，并且经常是 用阿片类镇痛药治疗。但是，阿片类药物的有效性最低，经常被遗忘。发展 我们对集中式疼痛的理解有限的理解阻碍了非阿片类镇痛药 表型与细胞水平的变化有关。在研究环境中，通常评估集中式疼痛 使用定量感觉测试（QST）。但是，QST耗时，需要大量的评估者 训练，容易出现测量误差。有关键的未满足需要开发可量化的测量 区分集中疼痛患者的细胞状态改变。必须解决这一需求 实现我们的长期目标，即为患有 风湿病。这个高风险，高回报的建议的目的是为 RA患者的外周血单核细胞（PBMC）与集中疼痛之间的关系。 我们的提议强调了使用多参数流式细胞术和 单细胞RNA测序（SCRNA-SEQ），以识别与PBMC的特征 DMARD治疗后集中疼痛。我们的中心假设是组成和基因的差异 循环免疫球的表达，尤其是单核细胞，将表征具有集中式的RA患者 疼痛表型与没有集中疼痛表型的表型。在AIM 1中，我们将确定协会 PBMC的免疫表型和集中式疼痛之间。我们将注册50名RA患者 最小的关节炎症，但DMARD治疗后疼痛的水平有所不同。这些患者将经历QST 测试由斜方肌肌肉的压痛阈值评估的集中疼痛是否相关 用：a）循环中每种细胞类型的比例，b）每种细胞的激活状态的连续度量 细胞类型。在AIM 2中，我们将确定循环PBMC的转录异质性的差异 在具有集中疼痛表型的患者与没有集中疼痛表型的患者之间。我们 将对来自最高（n = 10）和最低（n = 10）水平的AIM 1的患者进行SCRNA-SEQ 集中疼痛。我们将研究转录亚群和细胞类型特异性基因的差异 两组之间的表达。我们的建议具有很高的影响潜力，因为如果成功，它将提供 对RA患者的循环PBMC在改变疼痛调节中的作用的新颖见解，原型 系统性炎症状况。该提案的数据将用于通知R01应用程序以识别 RA患者集中疼痛的可再现外周血生物标志物。