Establishing a Single-Cell Proteomic Atlas for Normal and Osteoarthritic Articular Cartilage

建立正常和骨关节炎关节软骨的单细胞蛋白质组图谱

• 批准号: 10612005
• 负责人: Nidhi Bhutani
• 金额: $ 53.27万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-17 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612005
• 关键词: Ablation; Acceleration; Affect; Animal Model; Animals; Antibodies; Atlases; Cartilage; Cartilage Diseases; Cells; Chondrocytes; Clinical Trials; Cytometry; Data; Degenerative polyarthritis; Detection; Disease; Disease Progression; Encapsulated; Event; Extracellular Matrix; Failure; Goals; Heterogeneity; Homeostasis; Human; I-kappa B Proteins; In Situ; Individual; Inflammation; Inflammatory; Joints; Knowledge; Label; Maps; Medical; Molecular; Pain management; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Population; Proteins; Proteomics; Publishing; Rare Earth Metals; Replacement Arthroplasty; Reporting; Research; Resolution; Resources; Sampling; Science; Signal Pathway; Stains; Techniques; Technology; Testing; Therapeutic; Tissue Engineering; Tissues; Traumatic Arthropathy; articular cartilage; cartilage regeneration; cartilage repair; cell type; cohort; drug candidate; drug development; inflammatory marker; inflammatory modulation; inhibitor; insight; mouse model; novel; pre-clinical; receptor; regeneration potential; regenerative; regenerative cell; response; senescence; single-cell RNA sequencing; small molecule inhibitor; success; therapeutic evaluation; transcriptome sequencing; transcriptomics

Abstract Although multiple pathways and targets have been proposed for OA treatment, the rate of drug failure in clinical trials has been astoundingly high. The reasons for the limited success include the late detection of the disease and a lack of understanding of the molecular heterogeneity between patients. In this proposal, we aim to capitalize on the newly developed single-cell proteomic technique, mass cytometry (CyTOF) that allows detection of 40-80 proteins simultaneously in single cells, with the aim of identifying the diverse cellular subpopulations in OA cartilage. Although cartilage is a relatively simple tissue, with a single cell type being encapsulated in its secreted extracellular matrix (ECM), the variable degree of degeneration associated with each OA patient suggests that understanding this tissue (and other joint tissues) at a single cell level can provide novel insights into both OA pathology and patient heterogeneity. This will compliment single-cell transcriptomic data, with the additional advantage that the proteomic snapshot can also identify active signaling pathways in the identified subpopulations. The single-cell proteomic approach is especially pertinent in robustly identifying rare cell populations that are difficult to discern from RNA-sequencing data. In this proposal, we will establish single cell profiles of a large cohort of OA cartilage samples using a refined panel of rare earth metal labeled antibodies in Aim1 to identify distinct subpopulations in OA cartilage. In aim 2, we will test if the modulation of two newly identified rare subpopulations would be therapeutic in a mouse model of post-traumatic OA as well as follow their dynamics with disease progression. In Aim 3, we will analyze how drug treatments affect the cartilage subpopulations and their crosstalk in different patients especially to discern between a uniform or heterogenous response among the patient cohort. Collectively, the proposed studies will be impactful in identifying novel regenerative and pathological cell populations in OA and testing the therapeutic potential of their modulation.

抽象的 尽管针对 OA 治疗提出了多种途径和靶点，但临床上的药物失败率 试验率高得惊人。成功有限的原因包括疾病发现较晚 以及缺乏对患者之间分子异质性的了解。在本提案中，我们的目标是 利用新开发的单细胞蛋白质组学技术，质谱流式细胞仪 (CyTOF)，允许 同时检测单个细胞中的 40-80 个蛋白质，目的是识别不同的细胞 OA 软骨中的亚群。尽管软骨是一种相对简单的组织，具有单一细胞类型 封装在其分泌的细胞外基质（ECM）中，与相关的不同程度的退化 每个 OA 患者都认为，在单细胞水平上了解这种组织（和其他关节组织）可以提供 对 OA 病理学和患者异质性的新颖见解。这将补充单细胞转录组学 数据，还有一个额外的优点，即蛋白质组快照还可以识别活跃的信号通路 已确定的亚群。单细胞蛋白质组学方法对于可靠地识别 难以从 RNA 测序数据中辨别的稀有细胞群。在本提案中，我们将建立 使用稀土金属标记的精制面板对大量 OA 软骨样本进行单细胞分析 Aim1 中的抗体可识别 OA 软骨中的不同亚群。在目标 2 中，我们将测试是否调制 两个新发现的罕见亚群可用于治疗创伤后 OA 小鼠模型 跟踪它们随疾病进展的动态。在目标 3 中，我们将分析药物治疗如何影响软骨 亚群及其在不同患者中的串扰，尤其是区分均匀或异质的 患者队列中的反应。总的来说，拟议的研究将对识别新颖的 OA 中的再生和病理细胞群并测试其调节的治疗潜力。