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

基本信息

批准号：
10405629
负责人：
Nidhi Bhutani
金额：
$ 52.74万
依托单位：
STANFORD UNIVERSITY
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：
2021-05-17 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10405629
关键词：
Affect Animal Model Animals Antibodies Atlases Cartilage Cartilage Diseases Cells Chondrocytes Clinical Trials Cytometry D Cells 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 Reporting Research Resolution Resources Sampling Science Signal Pathway Stains Techniques Technology Testing Therapeutic Tissue Engineering Tissues articular cartilage cartilage regeneration cartilage repair cell type cohort drug candidate drug development inflammatory marker 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

项目摘要

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.

抽象的尽管已经提出了多种途径和靶标进行OA处理，但临床药物衰竭率试验一直很高。成功有限的原因包括对疾病的晚期发现并且缺乏对患者之间分子异质性的了解。在此提案中，我们的目标是利用新开发的单细胞蛋白质组学技术，质量细胞术（CYTOF），该技术允许单个细胞中同时检测40-80蛋白，目的是识别多种细胞 OA软骨中的亚群。尽管软骨是一种相对简单的组织，但单细胞类型为封装在其分泌的细胞外基质（ECM）中，变性程度与每个OA患者都建议在单个细胞水平上了解该组织（和其他关节组织）可以提供对OA病理和患者异质性的新颖见解。这将补充单细胞转录组数据，具有额外优势，即蛋白质组学快照还可以识别活动信号通路确定的亚群。单细胞蛋白质组学方法在鲁棒性识别中尤其相关罕见的细胞群很难与RNA测序数据辨别。在此提案中，我们将建立使用精制的稀土金属标记的大量OA软骨样品的单细胞剖面 AIM1中的抗体以鉴定OA软骨中的不同亚群。在AIM 2中，我们将测试是否调制在创伤后的OA和遵循他们的动力，疾病进展。在AIM 3中，我们将分析药物治疗如何影响软骨在不同患者中的亚群及其串扰，特别是辨别均匀或异质的患者队列中的反应。总体而言，拟议的研究将对识别新颖 OA中的再生和病理细胞群，并测试其调节的治疗潜力。