title: Preclinical safety and efficacy of human CD34(+) cells transduced with lentiviral vector for the treatment of Wiskott-Aldrich syndrome.
abstract: Gene therapy with ex vivo-transduced hematopoietic stem/progenitor cells may represent a valid therapeutic option for monogenic immunohematological disorders such as Wiskott-Aldrich syndrome (WAS), a primary immunodeficiency associated with thrombocytopenia. We evaluated the preclinical safety and efficacy of human CD34(+) cells transduced with lentiviral vectors (LV) encoding WAS protein (WASp). We first set up and validated a transduction protocol for CD34(+) cells derived from bone marrow (BM) or mobilized peripheral blood (MPB) using a clinical grade, highly purified LV. Robust transduction of progenitor cells was obtained in normal donors and WAS patients' cells, without evidence of toxicity. To study biodistribution of human cells and exclude vector release in vivo, LV-transduced CD34(+) cells were transplanted in immunodeficient mice, showing a normal engraftment and differentiation ability towards transduced lymphoid and myeloid cells in hematopoietic tissues. Vector mobilization to host cells and transmission to germline cells of the LV were excluded by different molecular assays. Analysis of vector integrations showed polyclonal integration patterns in vitro and in human engrafted cells in vivo. In summary, this work establishes the preclinical safety and efficacy of human CD34(+) cells gene therapy for the treatment of WAS.
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy, Milan, Italy.
authors.forename: Samantha
authors.initials: S
authors.lastname: Scaramuzza
authors.forename: Luca
authors.initials: L
authors.lastname: Biasco
authors.forename: Anna
authors.initials: A
authors.lastname: Ripamonti
authors.forename: Maria C
authors.initials: MC
authors.lastname: Castiello
authors.forename: Mariana
authors.initials: M
authors.lastname: Loperfido
authors.forename: Elena
authors.initials: E
authors.lastname: Draghici
authors.forename: Raisa J
authors.initials: RJ
authors.lastname: Hernandez
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Benedicenti
authors.forename: Marina
authors.initials: M
authors.lastname: Radrizzani
authors.forename: Monica
authors.initials: M
authors.lastname: Salomoni
authors.forename: Marco
authors.initials: M
authors.lastname: Ranzani
authors.forename: Cynthia C
authors.initials: CC
authors.lastname: Bartholomae
authors.forename: Elisa
authors.initials: E
authors.lastname: Vicenzi
authors.forename: Andrea
authors.initials: A
authors.lastname: Finocchi
authors.forename: Robbert
authors.initials: R
authors.lastname: Bredius
authors.forename: Marita
authors.initials: M
authors.lastname: Bosticardo
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Christof
authors.initials: C
authors.lastname: von Kalle
authors.forename: Eugenio
authors.initials: E
authors.lastname: Montini
authors.forename: Alessandra
authors.initials: A
authors.lastname: Biffi
authors.forename: Maria G
authors.initials: MG
authors.lastname: Roncarolo
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
authors.forename: Anna
authors.initials: A
authors.lastname: Villa
authors.forename: Alessandro
authors.initials: A
authors.lastname: Aiuti
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D016026:Bone Marrow Transplantation; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D018345:Mice, Knockout; D014161:Transduction, Genetic; D014923:Wiskott-Aldrich Syndrome / Q000628:therapy*
chemical_list: D018952:Antigens, CD34