Glycoproteomics Analysis to Identify Potential Biomarkers and Investigate CD14 Monocyte Activation Mechanisms via Exosomal Proteins in Cerebral Infarction Using a Novel Glycogen-Functionalized Nanoprobe.

Journal: Analytical chemistry
Published Date: Jul 3, 2025

Abstract

Cerebral infarction (CI) is a leading cause of disability and mortality, with activated plasma monocytes and altered protein glycosylation identified as critical contributors to its pathology. However, the mechanisms linking peripheral monocyte activation and glycoproteins in CI remain inadequately understood. In this study, we developed a glycogen-functionalized nanoprobe (mSiO@FeO@Glycogen) to profile plasma N-glycosylated proteins in healthy controls, CI patients, and patients in the rehabilitation phase (CIR). We identified 13 plasma N-glycoproteins with increased expression in CI patients and reduced levels in CIR patients. These N-glycoproteins, enriched in extracellular exosomes and associated with processes such as blood coagulation, fibrin clot formation, and complement activation, were validated as potential biomarkers for distinguishing CI and CIR patients through machine learning models. Flow cytometry analysis of peripheral blood mononuclear cells (PBMCs) revealed a higher proportion of inflammatory CD14 monocytes and a lower proportion of anti-inflammatory CD16 monocytes in CI patients compared with CIR patients. To further investigate whether exosomal proteins contribute to peripheral monocyte inflammation and could exacerbate CI pathogenesis, we developed a two-dimensional size-exclusion chromatography (SEC) method to isolate pure plasma exosomes for proteomic analysis. Four specific N-glycoproteins carried by CI-derived exosomes were identified and demonstrated the ability to directly activate human monocytes, potentially amplifying inflammation at injury sites. Our study highlights the role of exosomal N-glycoproteins in CI pathogenesis and positions them as promising diagnostic biomarkers and therapeutic targets for cerebral infarction.

Authors

  • Xiao Tong
    Shanghai Key Laboratory of Metabolic Remodeling and Health, Institute of Metabolism & Integrative Biology, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200438, China.
  • Weiwei Chen
    Department of Developmental and Behavioral Pediatrics, Shanghai Children's Medical Center affiliated to Shanghai Jiaotong University School of Medicine, Ministry of Education-Shanghai Key Laboratory of Children's Environmental Health, Shanghai, China.
  • Yan Xia
    Radiological Sciences Lab, Stanford University, 94305, CA, USA.
  • Yanmei Wang
    CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, PR China. Electronic address: wangyanm@ustc.edu.cn.
  • Shuangshuang Liu
    School of Electronic and Information Engineering, Beijing Jiaotong University, Beijing 100044, China.
  • Yingli Jia
    Shanghai Key Laboratory of Metabolic Remodeling and Health, Institute of Metabolism & Integrative Biology, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200438, China.
  • Jiaxi Wang
    Key Laboratory of Optoelectronic Technology and System of the Education Ministry of China, Chongqing University, Chongqing 400044, People's Republic of China. Engineering Research Center of Industrial Computed Tomography Nondestructive Testing of the Education Ministry of China, Chongqing University, Chongqing 400044, People's Republic of China.
  • Li-Hao Huang
    Shanghai Key Laboratory of Metabolic Remodeling and Health, Institute of Metabolism & Integrative Biology, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200438, China.

Keywords

External Resources

Popular Topics
Recent Journals