摘要
背景:椎间盘退变是下腰痛最常见的原因,然而其发病机制尚不完全清楚,因此探索预防和治疗椎间盘退变的新策略势在必行。单细胞转录组测序技术可以在单个细胞水平对mRNA进行高通量测序,揭示细胞的异质性和特定亚群,深入了解椎间盘退变发生发展的调控机制,提高对于潜在靶点的识别,促进早期诊断和治疗的探索。目的:了解单细胞转录组测序技术及其主要流程并综述近几年单细胞转录组测序技术在椎间盘退变中的研究进展,探讨其在发现靶向生物标志物方面的应用潜力。方法:应用计算机检索PubMed、Web of Science、中国知网、万方数据库中2014年1月至2024年6月发表的相关文献,中文检索词为“单细胞转录组测序,测序技术,椎间盘退变,髓核,纤维环,软骨终板”,英文检索词为“single-cell RNA sequencing,sequencing technology,intervertebral disc degeneration,nucleus pulposus,annulus fibrosus,cartilage endplate”,排除重复性文献和非相关文献,共筛选出57篇文献进行综述分析。结果与结论:①单细胞转录组测序技术已成为研究单细胞水平基因表达的强有力工具,该技术在椎间盘退变中揭示了新的亚群及其功能,提高了对椎间盘退变病理过程的理解;识别了髓核中的几种新型细胞亚群,包括肥大软骨样髓核细胞、效应髓核细胞和稳态髓核细胞等,阐明了这些细胞亚群在椎间盘退变过程中所发挥的功能及分化路径,在椎间盘退变的不同阶段,这些细胞亚群的比例及主导群体有所不同;发现了新的纤维环细胞亚群Grem1+亚群和Lum+亚群,Grem1+是纤维环的常驻祖亚群,Sox9和Id1是其关键调控因子,与组织发育和细胞分化有关;Nr2f2和Creb5可能负责Lum+亚群的血管化功能,为椎间盘退变治疗和组织修复提供了潜在的细胞来源和调控靶点。②在软骨终板中发现的间充质软骨细胞可进一步细分为稳态、效应和调节性间充质软骨细胞,稳态软骨细胞主要参与骨化和胶原结合过程,调节软骨细胞主要参与刺激和控制反应,效应软骨细胞主要参与代谢过程。③退变椎间盘组织免疫细胞中的单核/巨噬细胞进一步富集为氧化应激、活化组织、稳态等单核/巨噬细胞,加深了对单核/巨噬细胞亚群在椎间盘退变进展过程中不同功能的理解,从而提供基于特定亚群的个性化治疗。总结了近年来单细胞转录组测序在椎间盘退变中的靶向生物标志物,为其诊断及治疗策略提供了新的见解。
BACKGROUND:Intervertebral disc degeneration is the most common cause of low back pain;however,its pathogenesis is not yet fully understood.Therefore,it is imperative to explore new strategies for the prevention and treatment of intervertebral disc degeneration.Single-cell RNA sequencing allows highthroughput sequencing of mRNAs at the single-cell level,revealing cell heterogeneity and specific subgroups.This technology provides deeper insights into the regulatory mechanisms of the development of intervertebral disc degeneration,enhances the identification of potential targets,and promotes the exploration of early diagnosis and treatment.OBJECTIVE:To understand single-cell RNA sequencing and its main processes,and to review the recent research progress in single-cell RNA sequencing in intervertebral disc degeneration over the past few years,exploring its potential applications in the discovery of targeted biomarkers.METHODS:A computerized search of relevant literature published from January 2014 to June 2024 in PubMed,Web of Science,CNKI,and WanFang databases was performed using the search terms“single-cell RNA sequencing,sequencing technology,intervertebral disc degeneration,nucleus pulposus,annulus fibrosus,cartilage endplate”in English and Chinese.A total of 57 articles were finally selected for relevant analysis after duplicate and irrelevant literature was excluded.RESULTS AND CONCLUSION:(1)Single-cell RNA sequencing has emerged as a powerful tool for studying gene expression at the single-cell level.It has identified new cell subpopulations and elucidated their functions in intervertebral disc degeneration,thereby enhancing our understanding of the pathological processes involved in this condition.Several new cell subpopulations in the nucleus pulposus have been identified,including hypertrophic cartilage-like nucleus pulposus cells,effector nucleus pulposus cells,and homeostatic nucleus pulposus cells.These findings elucidate the functions and differentiation pathways of these subpopulations during intervertebral disc degeneration,highlighting variations in their proportions and dominant groups at different stages of degeneration.New subpopulations of annulus fibrosus cells,including the Grem1+and Lum+subpopulations,have been discovered.Grem1+is a resident progenitor subpopulation in the annulus fibrosus,with Sox9 and Id1 as key regulatory factors involved in tissue development and cell differentiation.Nr2f2 and Creb5 may be responsible for the vascularization function of the Lum+subpopulation,providing potential cell sources and regulatory targets for the treatment and repair of intervertebral disc degeneration.(2)Mesenchymal chondrocytes found in the cartilage endplate can be further subdivided into homeostatic,effector,and regulatory subtypes.Homeostatic chondrocytes are primarily involved in ossification and collagen-binding processes,regulatory chondrocytes participate mainly in stimulatory and control responses,and effector chondrocytes are predominantly involved in metabolic processes.(3)Further enrichment of monocytes/macrophages in the immune cells of degenerating disc tissues into monocytes/macrophages for oxidative stress,activated tissues,and homeostasis has deepened the understanding of the different functions of monocyte/macrophage subpopulations during the progression of intervertebral disc degeneration to provide personalized treatments based on specific subpopulations.The review summarizes recent advances in single-cell RNA sequencing for identifying targeted biomarkers in intervertebral disc degeneration,offering new insights into diagnostic and therapeutic strategies.
作者
刘兆峰
杨学军
Liu Zhaofeng;Yang Xuejun(Inner Mongolia Medical University,Hohhot 010030,Inner Mongolia Autonomous Region,China;Spine Surgery Centre A,The Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010000,Inner Mongolia Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2025年第20期4351-4360,共10页
Chinese Journal of Tissue Engineering Research
关键词
单细胞转录组测序
椎间盘退变
椎间盘
髓核
纤维环
测序技术
高通量测序
细胞亚群
工程化组织构建
single-cell RNA sequencing
intervertebral disc degeneration
intervertebral disc
nucleus pulposus
annulus fibrosus
sequencing technology
highthroughput sequencing
cell subpopulation
engineered tissue construction
