The Proteomic Analysis of Chronic Migraine Exosomes Reveals Disease Patterns and Potential Biomarkers

Exosomes have been identified as optimal biomarkers to screen for multiple diseases. However, few studies focus on the abundant exosome population isolated from plasma of migraine. This study investigated whether proteins in abundant exosomes can aid in the diagnosis of chronic migraine (CM). Plasma exosomes were collected by centrifugation, from which protein samples were extracted. A pilot study (CM, 18; episodic migraine (EM), 26) followed by a second dataset (CM, 26; EM, 16; tension-type headache (TTH), 20; control, 22) was applied to establish a diagnostic model of CM. We employed proteomics based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) to search for potential candidate biomarkers in plasma exosomes from CM patients. In total, 530 proteins in plasma exosomes were co-detected. Among them, 13 proteins were found significantly dysregulated between the plasma exosomes of CM patients and other groups. The receiver operating characteristic curve analysis revealed a combination of six proteins (upregulated: RAP2B, AK1, BID, DAG1, PICALM, PSMB2) could distinguish CM patients with high accuracy. Linear correlation analysis showed that the combination was significantly correlated with Headache Impact Test (HIT-6) scores (assessing the negative impact of headaches on normal daily activity). The RT-qPCR results showed the same trends in CM models with nitroglycerin as the exosomal protein sequencing results. These data revealed dysregulated proteins in plasma exosomes of CM, and the combination of plasma exosomal proteins RAP2B, AK1, BID, DAG1, PICALM, and PSMB2 could serve as a novel candidate biomarker for CM diagnosis.