Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation

Abstract: β-thalassemia (β-thal) is a worldwide hereditary red cell disorder characterized by severe chronic anemia. Recently, the pyruvate kinase (PK) activator mitapivat has been shown to improve anemia and ineffective erythropoiesis in a mouse model of β-thal and in patients with non–transfusion-...

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Main Authors: Angela Siciliano, Angelo D’Alessandro, Alessandro Matte, Giovanni Bisello, Mariarita Bertoldi, Monika Dzieciatkowska, Amy Argabright, Richard Huot Pozzetto, Veronica Riccardi, Andrea Mattarei, Alberto Ongaro, Jacopo Ceolan, Roberta Russo, Leonardo Rivadeneyra, Megan Wind-Rotolo, Achille Iolascon, Carlo Brugnara, Lucia De Franceschi
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Blood Advances
Online Access:http://www.sciencedirect.com/science/article/pii/S2473952925001636
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author Angela Siciliano
Angelo D’Alessandro
Alessandro Matte
Giovanni Bisello
Mariarita Bertoldi
Monika Dzieciatkowska
Amy Argabright
Richard Huot Pozzetto
Veronica Riccardi
Andrea Mattarei
Alberto Ongaro
Jacopo Ceolan
Roberta Russo
Leonardo Rivadeneyra
Megan Wind-Rotolo
Achille Iolascon
Carlo Brugnara
Lucia De Franceschi
author_facet Angela Siciliano
Angelo D’Alessandro
Alessandro Matte
Giovanni Bisello
Mariarita Bertoldi
Monika Dzieciatkowska
Amy Argabright
Richard Huot Pozzetto
Veronica Riccardi
Andrea Mattarei
Alberto Ongaro
Jacopo Ceolan
Roberta Russo
Leonardo Rivadeneyra
Megan Wind-Rotolo
Achille Iolascon
Carlo Brugnara
Lucia De Franceschi
author_sort Angela Siciliano
collection DOAJ
description Abstract: β-thalassemia (β-thal) is a worldwide hereditary red cell disorder characterized by severe chronic anemia. Recently, the pyruvate kinase (PK) activator mitapivat has been shown to improve anemia and ineffective erythropoiesis in a mouse model of β-thal and in patients with non–transfusion-dependent thalassemia. Here, we showed that in vitro CD34+-derived erythroblasts from patients with β-thal (codb039) are characterized by persistent expression of 2 PK isoforms, PKR and PKM2, compared with healthy cells. Activation of PKR and PKM2 via mitapivat promoted significant metabolic reprogramming of β-thal erythroblasts, resulting in higher levels of high-energy phosphate compounds, including adenosine triphosphate (ATP) and triphosphate nucleoside pools. Proteomics analyses revealed an accumulation of PKR, suggesting a possible beneficial effect of mitapivat on the stability of PKs. Increased ATP availability was accompanied by a higher degree of protein phosphorylation, especially in proteins involved in cell cycle regulation at the transcriptional, translational, and posttranslational levels, supporting the effect of mitapivat on erythroid maturation. Upon treatment with mitapivat, β-thal erythroblasts showed decreased markers of oxidation, including cysteine oxidative posttranslational modifications, downregulation of heat shock protein 70 and peroxiredoxin-2 expression, and normalization of the redox-dependent subcellular distribution of the latter enzyme. Collectively, our data support a protective effect of mitapivat in β-thal erythropoiesis, an effect favored by its activation of persistently expressed PKR and PKM2. In addition to the anticipated benefits on energy metabolism, we report that mitapivat treatment mitigated the oxidative damage in β-thal erythropoiesis, ensuring improved β-thal erythroblast maturation and survival.
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spelling doaj-art-da048064392b4e308ce3fc8804534a5d2025-08-20T02:02:25ZengElsevierBlood Advances2473-95292025-06-019112818283010.1182/bloodadvances.2024013591Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidationAngela Siciliano0Angelo D’Alessandro1Alessandro Matte2Giovanni Bisello3Mariarita Bertoldi4Monika Dzieciatkowska5Amy Argabright6Richard Huot Pozzetto7Veronica Riccardi8Andrea Mattarei9Alberto Ongaro10Jacopo Ceolan11Roberta Russo12Leonardo Rivadeneyra13Megan Wind-Rotolo14Achille Iolascon15Carlo Brugnara16Lucia De Franceschi17Department of Engineering for Innovative Medicine, University of Verona, Verona, Italy; Azienda Ospedaliera Universitaria Integrata, Verona, ItalyDepartment of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CODepartment of Medicine, University of Verona, Verona, ItalyDepartment of Neuroscience, Biomedicine and Movement Sciences, University of Verona, Verona, ItalyDepartment of Neuroscience, Biomedicine and Movement Sciences, University of Verona, Verona, ItalyDepartment of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CODepartment of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CODepartment of Engineering for Innovative Medicine, University of Verona, Verona, ItalyDepartment of Engineering for Innovative Medicine, University of Verona, Verona, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, Padua, ItalyDepartment of Engineering for Innovative Medicine, University of Verona, Verona, Italy; Azienda Ospedaliera Universitaria Integrata, Verona, ItalyDipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Naples, Italy; CEINGE Biotecnologie Avanzate, Naples, ItalyAgios Pharmaceuticals, Inc, Cambridge, MAAgios Pharmaceuticals, Inc, Cambridge, MADipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli Federico II, Naples, Italy; CEINGE Biotecnologie Avanzate, Naples, ItalyDepartment of Laboratory Medicine, Boston Children’s Hospital, Boston, MA; Department of Pathology, Harvard Medical School, Boston, MADepartment of Engineering for Innovative Medicine, University of Verona, Verona, Italy; Azienda Ospedaliera Universitaria Integrata, Verona, Italy; Correspondence: Lucia De Franceschi, Department of Engineering for Innovation Medicine, University of Verona & AOUI Verona, P.Le L Scuro, 10, 37134 Verona, Italy;Abstract: β-thalassemia (β-thal) is a worldwide hereditary red cell disorder characterized by severe chronic anemia. Recently, the pyruvate kinase (PK) activator mitapivat has been shown to improve anemia and ineffective erythropoiesis in a mouse model of β-thal and in patients with non–transfusion-dependent thalassemia. Here, we showed that in vitro CD34+-derived erythroblasts from patients with β-thal (codb039) are characterized by persistent expression of 2 PK isoforms, PKR and PKM2, compared with healthy cells. Activation of PKR and PKM2 via mitapivat promoted significant metabolic reprogramming of β-thal erythroblasts, resulting in higher levels of high-energy phosphate compounds, including adenosine triphosphate (ATP) and triphosphate nucleoside pools. Proteomics analyses revealed an accumulation of PKR, suggesting a possible beneficial effect of mitapivat on the stability of PKs. Increased ATP availability was accompanied by a higher degree of protein phosphorylation, especially in proteins involved in cell cycle regulation at the transcriptional, translational, and posttranslational levels, supporting the effect of mitapivat on erythroid maturation. Upon treatment with mitapivat, β-thal erythroblasts showed decreased markers of oxidation, including cysteine oxidative posttranslational modifications, downregulation of heat shock protein 70 and peroxiredoxin-2 expression, and normalization of the redox-dependent subcellular distribution of the latter enzyme. Collectively, our data support a protective effect of mitapivat in β-thal erythropoiesis, an effect favored by its activation of persistently expressed PKR and PKM2. In addition to the anticipated benefits on energy metabolism, we report that mitapivat treatment mitigated the oxidative damage in β-thal erythropoiesis, ensuring improved β-thal erythroblast maturation and survival.http://www.sciencedirect.com/science/article/pii/S2473952925001636
spellingShingle Angela Siciliano
Angelo D’Alessandro
Alessandro Matte
Giovanni Bisello
Mariarita Bertoldi
Monika Dzieciatkowska
Amy Argabright
Richard Huot Pozzetto
Veronica Riccardi
Andrea Mattarei
Alberto Ongaro
Jacopo Ceolan
Roberta Russo
Leonardo Rivadeneyra
Megan Wind-Rotolo
Achille Iolascon
Carlo Brugnara
Lucia De Franceschi
Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation
Blood Advances
title Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation
title_full Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation
title_fullStr Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation
title_full_unstemmed Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation
title_short Mitapivat metabolically reprograms human β-thalassemic erythroblasts, increasing their responsiveness to oxidation
title_sort mitapivat metabolically reprograms human β thalassemic erythroblasts increasing their responsiveness to oxidation
url http://www.sciencedirect.com/science/article/pii/S2473952925001636
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