Articolo in rivista, 2019, ENG, 10.1016/j.jaci.2019.03.012

Lentiviral gene therapy corrects platelet phenotype and function in patients with Wiskott-Aldrich syndrome

Sereni L1, Castiello MC1, Di Silvestre D2, Della Valle P3, Brombin C4, Ferrua F5, Cicalese MP6, Pozzi L3, Migliavacca M6, Bernardo ME6, Pignata C7, Farah R8, Notarangelo LD9, Marcus N10, Cattaneo L11, Spinelli M12, Giannelli S1, Bosticardo M1, van Rossem K13, D'Angelo A3, Aiuti A5, Mauri P2, Villa A14.

1 San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy. 2 Proteomic and Metabolomic Laboratory, Institute of Biomedical Technologies, National Research Council (ITB-CNR), Segrate, Italy. 3 Coagulation Service & Thrombosis Research Unit, San Raffaele Scientific Institute, Milan, Italy. 4 University Centre for Statistics in the Biomedical Sciences (CUSSB), Vita-Salute San Raffaele University, Milan, Italy. 5 San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy; Vita-Salute San Raffaele University, Milan, Italy; Pediatric Immunohematology Unit, San Raffaele Scientific Institute, Milan, Italy. 6 San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy; Pediatric Immunohematology Unit, San Raffaele Scientific Institute, Milan, Italy. 7 Pediatric Section, Department of Translational Medical Sciences, University of Naples Federico II, Naples, Italy. 8 Department of Pediatrics, Division of Hematology-Oncology, Saint George Hospital University Medical Centre, Beirut, Lebanon. 9 Pediatric Onco-Haematology and BMT Unit, Children's Hospital, ASST Spedali Civili of Brescia, Brescia, Italy. 10 Department of Pediatrics, Schneider Children's Medical Center of Israel, Petach Tikva, Israel; Kipper Institute of Immunology, Schneider Children's Medical Center of Israel, Petach Tikva, Israel; Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel. 11 SC Pediatria, Ospedale Infantile C. Arrigo, Alessandria, Italy. 12 Pediatric Clinic, MBBM Foundation, Maria Letizia Verga Center, Monza, Italy. 13 Rare Diseases Unit, GlaxoSmithKline, Brentford, United Kingdom. 14 San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy; Milan Unit, Istituto di Ricerca Genetica e Biomedica, Consiglio Nazionale delle Ricerche, Milan, Italy. Electronic address: villa.anna@hsr.it.

BACKGROUND: Thrombocytopenia is a serious issue for all patients with classical Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT) because it causes severe and life-threatening bleeding. Lentiviral gene therapy (GT) for WAS has shown promising results in terms of immune reconstitution. However, despite the reduced severity and frequency of bleeding events, platelet counts remain low in GT-treated patients. OBJECTIVE: We carefully investigated platelet defects in terms of phenotype and function in untreated patients with WAS and assessed the effect of GT treatment on platelet dysfunction. METHODS: We analyzed a cohort of 20 patients with WAS/XLT, 15 of them receiving GT. Platelet phenotype and function were analyzed by using electron microscopy, flow cytometry, and an aggregation assay. Platelet protein composition was assessed before and after GT by means of proteomic profile analysis. RESULTS: We show that platelets from untreated patients with WAS have reduced size, abnormal ultrastructure, and a hyperactivated phenotype at steady state, whereas activation and aggregation responses to agonists are decreased. GT restores platelet size and function early after treatment and reduces the hyperactivated phenotype proportionally to WAS protein expression and length of follow-up. CONCLUSIONS: Our study highlights the coexistence of morphologic and multiple functional defects in platelets lacking WAS protein and demonstrates that GT normalizes the platelet proteomic profile with consequent restoration of platelet ultrastructure and phenotype, which might explain the observed reduction of bleeding episodes after GT. These results are instrumental also from the perspective of a future clinical trial in patients with XLT only presenting with microthrombocytopenia.

Journal of allergy and clinical immunology 144 (3), pp. 825–838

Keywords

Wiskott-Aldrich syndrome, X-linked thrombocytopenia, gene therapy, platelets

CNR authors

Di Silvestre Dario, Villa Anna, Mauri Pietro Luigi

CNR institutes

IRGB – Istituto di Ricerca Genetica e Biomedica, ITB – Istituto di tecnologie biomediche

ID: 417797

Year: 2019

Type: Articolo in rivista

Creation: 2020-02-28 16:31:52.000

Last update: 2022-06-13 15:48:39.000

External links

OAI-PMH: Dublin Core

OAI-PMH: Mods

OAI-PMH: RDF

DOI: 10.1016/j.jaci.2019.03.012

External IDs

CNR OAI-PMH: oai:it.cnr:prodotti:417797

DOI: 10.1016/j.jaci.2019.03.012

ISI Web of Science (WOS): 000485222300026