Targeted Metabolomics Highlights Dramatic Antioxidant Depletion, Increased Oxidative/Nitrosative Stress and Altered Purine and Pyrimidine Concentrations in Serum of Primary Myelofibrosis Patients

Renata Mangione; Cesarina Giallongo; Andrea Duminuco; Enrico La Spina; Lucia Longhitano; Sebastiano Giallongo; Daniele Tibullo; Giuseppe Lazzarino; Miriam Wissam Saab; Arianna Sbriglione; Giuseppe A Palumbo; Andrea Graziani; Amer M Alanazi; Valentina Di Pietro; Barbara Tavazzi; Angela Maria Amorini; Giacomo Lazzarino

doi:10.3390/antiox13040490

Targeted Metabolomics Highlights Dramatic Antioxidant Depletion, Increased Oxidative/Nitrosative Stress and Altered Purine and Pyrimidine Concentrations in Serum of Primary Myelofibrosis Patients

Antioxidants (Basel). 2024 Apr 19;13(4):490. doi: 10.3390/antiox13040490.

Authors

Renata Mangione^{1

2}, Cesarina Giallongo³, Andrea Duminuco⁴, Enrico La Spina⁵, Lucia Longhitano⁵, Sebastiano Giallongo³, Daniele Tibullo⁵, Giuseppe Lazzarino⁵, Miriam Wissam Saab⁵, Arianna Sbriglione⁵, Giuseppe A Palumbo³, Andrea Graziani², Amer M Alanazi⁶, Valentina Di Pietro^{7

8}, Barbara Tavazzi², Angela Maria Amorini⁵, Giacomo Lazzarino²

Affiliations

¹ Department of Basic Biotechnological Sciences, Intensive and Perioperative Clinics, Catholic University of the Sacred Heart of Rome, Largo F. Vito 1, 00168 Rome, Italy.
² Departmental Faculty of Medicine, UniCamillus-Saint Camillus International University of Health and Medical Sciences, Via di Sant'Alessandro 8, 00131 Rome, Italy.
³ Department of Medical and Surgical Sciences and Advanced Technologies "G.F. Ingrassia", Division of Hematology, University of Catania, Via S. Sofia 87, 95123 Catania, Italy.
⁴ Hematology Unit with BMT, A.O.U. Policlinico "G.Rodolico-San Marco", Via S. Sofia 78, 95123 Catania, Italy.
⁵ Department of Biomedical and Biotechnological Sciences, Division of Medical Biochemistry, University of Catania, Via S. Sofia 97, 95123 Catania, Italy.
⁶ Pharmaceutical Biotechnology Laboratory, Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia.
⁷ Neurotrauma and Ophthalmology Research Group, School of Clinical and Experimental Medicine, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
⁸ National Institute for Health Research Surgical Reconstruction and Microbiology Research Centre, Queen Elizabeth Hospital, Edgbaston, Birmingham B15 2TH, UK.

Abstract

To date, little is known concerning the circulating levels of biochemically relevant metabolites (antioxidants, oxidative/nitrosative stress biomarkers, purines, and pyrimidines) in patients with primary myelofibrosis (PMF), a rare form of myeloproliferative tumor causing a dramatic decrease in erythropoiesis and angiogenesis. In this study, using a targeted metabolomic approach, serum samples of 22 PMF patients and of 22 control healthy donors were analyzed to quantify the circulating concentrations of hypoxanthine, xanthine, uric acid (as representative purines), uracil, β-pseudouridine, uridine (as representative pyrimidines), reduced glutathione (GSH), ascorbic acid (as two of the main water-soluble antioxidants), malondialdehyde, nitrite, nitrate (as oxidative/nitrosative stress biomarkers) and creatinine, using well-established HPLC method for their determination. Results showed that PMF patients have dramatic depletions of both ascorbic acid and GSH (37.3- and 3.81-times lower circulating concentrations, respectively, than those recorded in healthy controls, p < 0.0001), accompanied by significant increases in malondialdehyde (MDA) and nitrite + nitrate (4.73- and 1.66-times higher circulating concentrations, respectively, than those recorded in healthy controls, p < 0.0001). Additionally, PMF patients have remarkable alterations of circulating purines, pyrimidines, and creatinine, suggesting potential mitochondrial dysfunctions causing energy metabolism imbalance and consequent increases in these cell energy-related compounds. Overall, these results, besides evidencing previously unknown serum metabolic alterations in PMF patients, suggest that the determination of serum levels of the aforementioned compounds may be useful to evaluate PMF patients on hospital admission for adjunctive therapies aimed at recovering their correct antioxidant status, as well as to monitor patients' status and potential pharmacological treatments.

Keywords: GSH; HPLC; antioxidants; ascorbic acid; oxidative/nitrosative stress; primary myelofibrosis; purines; pyrimidines; serum.

Grants and funding

The authors Daniele Tibullo, Giuseppe Lazzarino, Angela Maria Amorini, were supported by the University Research Project Grant (PIACERI 2020–2022), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, Italy. The author Amer M. Alanazi was funded by the researchers supporting project number (RSP2024R261) King Saud University, Riyadh, Saudi Arabia.