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

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY

卷:144

Article

Sereni, Lucia¹  Castiello, Maria Carmina¹  Di Silvestre, Dario⁵  Della Valle, Patrizia^2,3  Brombin, Chiara⁶  Ferrua, Francesca^1,4,7  Cicalese, Maria Pia^1,4  Pozzi, Loris^2,3  Migliavacca, Maddalena^1,4  Bernardo, Maria Ester^1,4  Pignata, Claudio⁸  Farah, Roula⁹  Notarangelo, Lucia Dora¹⁰  Marcus, Nufar^11,12,13  Cattaneo, Lorella¹⁴  Spinelli, Marco¹⁵  Giannelli, Stefania¹  Bosticardo, Marita^1,18  van Rossem, Koen¹⁶  D'Angelo, Armando^2,3  Aiuti, Alessandro^1,4,7  Mauri, Pierluigi⁵  Villa, Anna^1,17 

[1] IRCCS San Raffaele Sci Inst, San Raffaele Telethon Inst Gene Therapy SR Tiget, Div Regenerat Med Stem Cells & Gene Therapy, Milan, Italy

[2] IRCCS San Raffaele Sci Inst, Coagulat Serv, Milan, Italy

[3] IRCCS San Raffaele Sci Inst, Thrombosis Res Unit, Milan, Italy

[4] IRCCS San Raffaele Sci Inst, Pediat Immunohematol Unit, Milan, Italy

[5] CNR, ITB, Natl Res Council, Prote & Metabol Lab,Inst Biomed Technol, Segrate, Italy

[6] Univ Vita Salute San Raffaele, Univ Ctr Stat Biomed Sci CUSSB, Milan, Italy

[7] Univ Vita Salute San Raffaele, Milan, Italy

[8] Univ Naples Federico II, Dept Translat Med Sci, Pediat Sect, Naples, Italy

[9] St George Hosp Univ Med Ctr, Dept Pediat, Div Hematol Oncol, Beirut, Lebanon

[10] Childrens Hosp, ASST Spedali Civili Brescia, Pediat Oncohaematol & BMT Unit, Brescia, Italy

[11] Schneider Childrens Med Ctr Israel, Dept Pediat, Petah Tiqwa, Israel

[12] Schneider Childrens Med Ctr Israel, Kipper Inst Immunol, Petah Tiqwa, Israel

[13] Tel Aviv Univ, Sackler Fac Med, Tel Aviv, Israel

[14] Osped Infantile C Arrigo, SC Pediat, Alessandria, Italy

[15] MBBM Fdn, Maria Letizia Verga Ctr, Pediat Clin, Monza, Italy

[16] GlaxoSmithKline, Rare Dis Unit, Brentford, England

[17] CNR, Ist Ric Genet & Biomed, Milan Unit, Milan, Italy

[18] Natl Inst Allergy & Infect Dis, Lab Clin Immunol & Microbiol, IDGS, Div Intramural Res,NIH,Dept Hlth & Human Serv, Bethesda, MD USA

关键词: Wiskott-Aldrich syndrome;    X-linked thrombocytopenia;    gene therapy;    platelets;   

DOI  :  10.1016/j.jaci.2019.03.012

来源: Elsevier

【 摘 要 】

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.

【 授权许可】

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