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      Quantitative proteomics of plasma vesicles identify novel biomarkers for hemoglobin E/β-thalassemic patients.

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          Hemoglobin E (HbE)/β-thalassemia has a wide spectrum of clinical manifestations that cannot be explained purely by its genetic background. Circulating extracellular vesicles (EVs) are one factor that likely contributes to disease severity. This study has explored the differences in protein composition and quantity between EVs from HbE/β-thalassemic patients and healthy individuals. We used tandem mass tag labeling mass spectrometry to analyze the EV proteins isolated from the plasma of 15 patients compared with the controls. To reduce biological variation between individuals, the EV proteins isolated from randomly assigned groups of 5 HbE/β-thalassemic patients were pooled and compared with 5 pooled age- and sex-matched controls in 3 separate experiments. Alpha hemoglobin-stabilizing protein had the highest fold increase. Catalase, superoxide dismutase, T-complex proteins, heat shock proteins, transferrin receptor, ferritin, and cathepsin S were also upregulated in thalassemic circulating EVs. Importantly, haptoglobin and hemopexin were consistently reduced in patients' EVs across all data sets, in keeping with the existing hemolysis that occurs in thalassemia. The proteomic data analysis of EV samples isolated from 6 individual HbE/β-thalassemic patients and western blotting results corroborated these findings. In conclusion, we have successfully identified consistent alterations of protein quantity between EVs from HbE/β-thalassemic and healthy individuals. This work highlights haptoglobin, hemopexin, and cathepsin S as potential clinically relevant biomarkers for levels of hemolysis and inflammation. Monitoring of these plasma proteins could help in the clinical management of thalassemia.

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          Author and article information

          Blood Adv
          Blood advances
          American Society of Hematology
          Jan 23 2018
          : 2
          : 2
          [1 ] Department of Transfusion Medicine, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand.
          [2 ] Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant, Bristol, United Kingdom.
          [3 ] School of Biochemistry and.
          [4 ] Proteomics Facility, University of Bristol, Bristol, United Kingdom.
          [5 ] Department of Medicine Faculty of Medicine Siriraj Hospital, Bangkok, Thailand; and.
          [6 ] National Institute for Health Research Blood and Transplant Unit in Red Blood Cell Products, University of Bristol, Bristol, United Kingdom.


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