Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Jesús Castro-Marrero, Esther Serrano-Pertierra, Myriam Oliveira-Rodríguez, Maria Cleofé Zaragozá, Alba Martínez-Martínez, María Del Carmen Blanco-López, José Alegre, Jesús Castro-Marrero, Esther Serrano-Pertierra, Myriam Oliveira-Rodríguez, Maria Cleofé Zaragozá, Alba Martínez-Martínez, María Del Carmen Blanco-López, José Alegre

Abstract

Chronic Fatigue Syndrome (CFS), also known as Myalgic Encephalomyelitis (ME) is an acquired, complex and multisystem condition of unknown etiology, no established diagnostic lab tests and no universally FDA-approved drugs for treatment. CFS/ME is characterised by unexplicable disabling fatigue and is often also associated with numerous core symptoms. A growing body of evidence suggests that extracellular vesicles (EVs) play a role in cell-to-cell communication, and are involved in both physiological and pathological processes. To date, no data on EV biology in CFS/ME are as yet available. The aim of this study was to isolate and characterise blood-derived EVs in CFS/ME. Blood samples were collected from 10 Spanish CFS/ME patients and 5 matched healthy controls (HCs), and EVs were isolated from the serum using a polymer-based method. Their protein cargo, size distribution and concentration were measured by Western blot and nanoparticle tracking analysis. Furthermore, EVs were detected using a lateral flow immunoassay based on biomarkers CD9 and CD63. We found that the amount of EV-enriched fraction was significantly higher in CFS/ME subjects than in HCs (p = 0.007) and that EVs were significantly smaller in CFS/ME patients (p = 0.014). Circulating EVs could be an emerging tool for biomedical research in CFS/ME. These findings provide preliminary evidence that blood-derived EVs may distinguish CFS/ME patients from HCs. This will allow offer new opportunities and also may open a new door to identifying novel potential biomarkers and therapeutic approaches for the condition.

Keywords: Chronic fatigue syndrome; extracellular vesicles; lateral flow immunoassay system; myalgic encephalomyelitis; tetraspanins.

Figures

Figure 1.
Figure 1.
(a) Box plot for the protein concentration of the EV fractions isolated in 10 CFS/ME patients and 5 healthy controls (HCs). (b) Box plot for the measurement of CD63 levels by optical density (O.D.) in CFS/ME individuals (n = 10) and HCs (n = 5). (c) Detection of CD63 (25 kDa) in EV fractions from CFS/ME patients and HCs by western blotting (left lane shows 26 kDa band corresponding to the molecular weight marker).
Figure 2.
Figure 2.
Characterization of isolated EVs by NTA. (a) Representative graph depicting the hydrodynamic size distribution profiles of isolated EVs from a CFS/ME subject and an HC, measured by NTA. (b) Box plots for the diameter sizes and (c) particle concentration of the EV fractions of study participants.
Figure 3.
Figure 3.
Detection of serum isolated-EVs using anti-CD9 as capture antibody and AuNP-anti-CD63 as detection probe. (a) Representative immunostrip test obtained of HCs and CFS/ME individuals. EV-depleted serum was used as negative control (C-). Control line (Ct). Test line (CD9). (b) Box plots for the signal intensities measured with the ESEQuant LR3 lateral flow strip reader of study participants.

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