Efficacy of gene therapy for Wiskott-Aldrich syndrome using a WAS promoter/cDNA-containing lentiviral vector and nonlethal irradiation.
Wiskott-Aldrich syndrome (WAS) is a life-threatening X-linked primary immunodeficiency characterized by infections, hemorrhages, autoimmune disorders, and lymphomas. Transplantation of genetically corrected autologous hematopoietic stem cells (HSCs) could represent an alternative treatment to allogeneic HSC transplantation, the latter being often associated with severe complications. We used WAS-/- mice to test the efficacy of a gene therapy approach based on nonlethal irradiation followed by transplantation of WAS-/- HSCs transduced with lentiviral vectors encoding the WAS protein (WASP) from either the ubiquitous PGK promoter or the tissue- specific WAS promoter. The procedure resulted in significant levels of engraftment of WASP-expressing T cells, B cells, platelets, and myeloid cells. T cells harbored one or two vector copies and displayed partial to full correction of T cell receptor-driven interleukin-2 production and proliferation. In addition, polymerization of F-actin and localization of WASP at the site of the immunological synapse were restored. The treatment was well tolerated and no pathology was detected by systematic blood analysis and autopsy. The efficacy of WAS gene transfer into HSCs, using the WAS promoter-containing lentiviral vector, combined with nonlethal irradiation provides a strong rationale for the development of gene therapy for WAS patients.
Dupré
Loïc
L
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), 20132 Milan, Italy.
Marangoni
Francesco
F
Scaramuzza
Samantha
S
Trifari
Sara
S
Hernández
Raisa Jofra
RJ
Aiuti
Alessandro
A
Naldini
Luigi
L
Roncarolo
Maria-Grazia
MG
D000199:Actins / Q000378:metabolism; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D049109:Cell Proliferation; D018076:DNA, Complementary / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D014161:Transduction, Genetic; D014182:Transplantation, Autologous; D014923:Wiskott-Aldrich Syndrome / Q000235:genetics / Q000628:therapy*; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics*
D000199:Actins; D018076:DNA, Complementary; D007376:Interleukin-2; D051316:Wiskott-Aldrich Syndrome Protein