Stefania Mondello,1 Firas H Kobeissy,2 Yehia Mechref,2 Jingfu Zhao,3 Samer El Hayek,4 Kazem Zibara,5 Monica Moresco,6,7 Giuseppe Plazzi,6 Filomena II Cosentino,8 Raffaele Ferri8 1Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, Messina, Italy; 2Faculty of Medicine, American University of Beirut Medical Center, Beirut, Lebanon; 3Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX, 79409, USA; 4Department of Psychiatry, American University of Beirut, Beirut, Lebanon; 5Department of Biology, Faculty of Sciences-I, PRASE, DSST, Lebanese University, Beirut, Lebanon; 6Department of Biomedical and Neuromotor Sciences, Alma Mater Studiorum, University of Bologna, Bologna, Italy; 7IRCCS Istituto delle Scienze Neurologiche di Bologna, Bologna, Italy; 8Sleep Research Centre, Oasi Research Institute - IRCCS, Troina, ItalyCorrespondence: Raffaele FerriSleep Research Centre, Oasi Research Institute - IRCCS, Troina, 94018, ItalyTel +39 0935 936111Fax +39 0935 936231Email rferri@oasi.en.itPurpose: We performed comparative proteomic analyses of blood of patients with RLS and healthy individuals aiming to identify potential biomarker and therapeutic target candidate for RLS.Patients and Methods: Blood serum samples from 12 patients with a clinical diagnosis of RLS (8 females and 4 males, with a mean age of 68.52 years) and 10 healthy controls (5 females and 5 males, with a mean age of 67.61 years) underwent proteomic profiling by liquid chromatography coupled with tandem mass spectrometry. Pathway analysis incorporating protein–protein interaction networks was carried out to identify pathological processes linked to the differentially expressed proteins.Results: We quantified 272 proteins in patients with RLS and healthy controls, of which 243 were shared. Five proteins – apolipoprotein C-II, leucine-rich alpha-2-glycoprotein 1, FLJ92374, extracellular matrix protein 1, and FLJ93143 – were substantially increased in RLS patients, whereas nine proteins – vitamin D-binding protein, FLJ78071, alpha-1-antitrypsin, CD5 antigen-like, haptoglobin, fibrinogen alpha chain, complement factor H-related protein 1, platelet factor 4, and plasma protease C1 inhibitor – were decreased. Bioinformatics analyses revealed that these proteins were linked to 1) inflammatory and immune response, and complement activation, 2) brain-related development, cell aging, and memory disorders, 3) pregnancy and associated complications, 4) myocardial infarction, and 5) reactive oxygen species generation and subsequent diabetes mellitus.Conclusion: Our findings shed light on the multifactorial nature of RLS and identified a set of circulating proteins that may have clinical importance as biomarkers and therapeutic targets.Keywords: idiopathic restless legs syndrome, biomarkers, LC-MS/MS, proteome, interactome
