色谱2023，Vol.41Issue(12)：1073-1083,11.DOI:10.3724/SP.J.1123.2022.12022
基于串联质量标记的帕金森病血浆及血浆外泌体定量蛋白质组学分析
Tandem mass tag-based quantitative proteomics analysis of plasma and plasma exosomes in Parkinson's disease
摘要
Abstract
The cardinal clinical features of Parkinson's disease(PD),a common neurodegen-erative disease,include the irreversible impairment of movement coordination,such as trem-ors,gait rigidity,bradykinesia,and hypokinesia.Although various factors are associated with the pathological changes in PD,such as oxidative stress,mitochondrial dysfunction,and neu-roinflammation,the availability of treatments to retard PD progression is limited.Therefore,novel biomarkers for PD diagnosis and therapeutic targets are urgently needed.The diagnosis of PD mainly depends on its clinical manifestations and has an error rate of approximately 20%.Studies have shown that α-synuclein(α-syn)levels are significantly increased in the cerebrospinal fluid of patients with PD;however,the invasive nature of lumbar puncture re-stricts further studies on its clinical applications.Hence,the development of novel peripheral blood markers would be helpful for the early diagnosis of PD.Exosomes are extracellular vesi-cles(EVs)released by various cell types under physiological and pathophysiological conditions.Because exosomes carry a variety of bioactive molecules,they play a key role in biological processes such as intercellular communication and the immune response.Central nervous sys-tem(CNS)-derived exosomes can be detected in the cerebrospinal and peripheral body fluids of patients with PD,and their contents are altered during the disease process,rendering them an attractive biomarker resource.Therefore,a comprehensive and high-throughput investigation of the plasma and its exosomes may enhance our understanding of PD.In this study,we isolated exosomes from plasma using standard differential centrifugation and performed tandem mass tag(TMT)-labeled quantitative proteomic analysis of plasma and plasma exosome samples from healthy individuals and patients with PD using liquid chromatography-tandem mass spec-trometry(LC-MS/MS).A total of 724 proteins were quantified in the plasma samples,and 611 proteins were screened from the exosome samples.Among these 611 proteins,413 were found in the Exosomal Protein Database(Exocarta).Using |log2 FC|＞0.26 and P-value(P)＜0.05 as the cutoff,five upregulated and six downregulated proteins were identified in the plasma sam-ples of the PD group compared with the healthy group.In the plasma exosome samples,com-pared with the healthy group,the PD group showed six upregulated and seven downregulated proteins.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analy-ses were conducted based on gene set enrichment analysis(GSEA).GO-cellular component(CC)analysis revealed that plasma-enriched proteins were mainly located in the nucleus whereas plasma exosome-enriched proteins were mainly located in the cytoplasm.According to the GO-molecular function(MF)analysis,the MFs of differentially expressed proteins in the plasma were mainly enriched in RNA,DNA binding,and complement binding.By contrast,the molecular functions of differentially expressed proteins derived from plasma exosomes were en-riched in antioxidant activity,oxidoreductase activity,and peroxide acceptor activity.We then analyzed the enriched KEGG pathways of differentially expressed proteins derived from the plasma and plasma exosome samples.The enrichment pathways of differentially expressed pro-teins in the plasma samples included the lysosome pathway,cellular senescence,and protein processing in the endoplasmic reticulum.By contrast,the enrichment pathways of differentially expressed proteins in the plasma exosome samples included chemokine signaling and cytokine receptor interactions.Finally,we assessed the functions of some exosomal proteins in PD to elucidate their potential for PD diagnosis and treatment.Significant differences were observed between the plasma and plasma exosome protein profiles,and the functions of differentially ex-pressed proteins in plasma exosomes were strongly related to the pathology of PD.Our study provides a reference for identifying the potential biomarkers and therapeutic targets of PD.关键词
串联质量标记/定量蛋白质组学/帕金森病/外泌体/血浆标志物Key words
tandem mass tag(TMT)/quantitative proteomics/Parkinson's disease(PD)/exosomes/plasma biomarkers分类
化学化工引用本文复制引用
赵媛,刘新,张译丹,张健,刘向,杨国锋..基于串联质量标记的帕金森病血浆及血浆外泌体定量蛋白质组学分析[J].色谱,2023,41(12):1073-1083,11.基金项目
河北省科技计划项目(22377798D). Science and Technology Program of Hebei Province(No.22377798D). （22377798D）
