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Foamy virus vector-mediated gene correction of a mouse model of Wiskott-Aldrich syndrome.

ABSTRACT: The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by eczema, thrombocytopenia and immunodeficiency. Hematopoietic cell transplantation can cure the disease and gene therapy is being tested as an alternative treatment option. In this study, we assessed the use of foamy virus (FV) vectors as a gene transfer system for WAS, using a Was knockout (KO) mouse model. Preliminary experiments using FV vectors expressing the green fluorescent protein under the transcriptional control of the endogenous WAS promoter or a ubiquitously acting chromatin opening element allowed us to define transduction conditions resulting in high (>40%) and long-term in-vivo marking of blood cells after transplantation. In following experiments, Was KO mice were treated with FV vectors containing the human WAS complementary DNA (cDNA). Transplanted animals expressed the WAS protein (WASp) in T and B lymphocytes, as well as platelets and showed restoration of both T-cell receptor-mediated responses and B-cell migration. We also observed recovery of platelet adhesion and podosome formation in dendritic cells (DCs) of treated mice. These data demonstrate that FV vectors can be effective for hematopoietic stem cell (HSC)-directed gene correction of WAS.

SUBMITTER: Uchiyama T

PROVIDER: S-EPMC3369282 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature

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Foamy virus vector-mediated gene correction of a mouse model of Wiskott-Aldrich syndrome.

Uchiyama Toru T Adriani Marsilio M Jagadeesh G Jayashree GJ Paine Adam A Candotti Fabio F

Molecular therapy : the journal of the American Society of Gene Therapy 20120103 6

The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by eczema, thrombocytopenia and immunodeficiency. Hematopoietic cell transplantation can cure the disease and gene therapy is being tested as an alternative treatment option. In this study, we assessed the use of foamy virus (FV) vectors as a gene transfer system for WAS, using a Was knockout (KO) mouse model. Preliminary experiments using FV vectors expressing the green fluorescent protein under the transcriptional control ...[more]

PMID: 22215016

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Project description:BackgroundWiskott-Aldrich syndrome (WAS) is a severe X-linked immunodeficiency characterized by microthrombocytopenia, eczema, recurrent infections, and susceptibility to autoimmunity and lymphomas. Hematopoietic stem cell transplantation is the treatment of choice; however, administration of WAS gene-corrected autologous hematopoietic stem cells has been demonstrated as a feasible alternative therapeutic approach.ObjectiveBecause B-cell homeostasis is perturbed in patients with WAS and restoration of immune competence is one of the main therapeutic goals, we have evaluated reconstitution of the B-cell compartment in 4 patients who received autologous hematopoietic stem cells transduced with lentiviral vector after a reduced-intensity conditioning regimen combined with anti-CD20 administration.MethodsWe evaluated B-cell counts, B-cell subset distribution, B cell-activating factor and immunoglobulin levels, and autoantibody production before and after gene therapy (GT). WAS gene transfer in B cells was assessed by measuring vector copy numbers and expression of Wiskott-Aldrich syndrome protein.ResultsAfter lentiviral vector-mediated GT, the number of transduced B cells progressively increased in the peripheral blood of all patients. Lentiviral vector-transduced progenitor cells were able to repopulate the B-cell compartment with a normal distribution of B-cell subsets both in bone marrow and the periphery, showing a WAS protein expression profile similar to that of healthy donors. In addition, after GT, we observed a normalized frequency of autoimmune-associated CD19(+)CD21(-)CD35(-) and CD21(low) B cells and a reduction in B cell-activating factor levels. Immunoglobulin serum levels and autoantibody production improved in all treated patients.ConclusionsWe provide evidence that lentiviral vector-mediated GT induces transgene expression in the B-cell compartment, resulting in ameliorated B-cell development and functionality and contributing to immunologic improvement in patients with WAS.

Project description:Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency characterized by microthrombocytopenia, eczema, recurrent infections, and increased incidence of autoimmune disorders and malignancies. WAS is caused by mutations in the was gene, which is expressed exclusively in hematopoietic cells, and the spleen plays an important role in hematopoiesis and red blood cell clearance. However, to date, detailed comparative analyses of the spleen between WASp-deficient (WAS-KO) mice and WT mice, particularly at the transcriptomic level, have not been reported. Here, we investigated the differences in the transcriptomes of spleen tissues of 10-week-old WAS-KO mice. Comparison of the gene expression profiles of WAS-KO and WT mice revealed 1964 differentially expressed genes (DEGs). Among these genes, 996 DEGs were upregulated, and 968 were downregulated in WAS-KO mice. To determine the functions of the DEGs, GO and KEGG enrichment analyses were performed for significantly upregulated and downregulated DEGs, respectively. The results showed that the levels of cell senescence and apoptosis-related genes were increased, that the antigen processing and presentation mechanisms involved in the immune response were damaged, and that signal transduction processes were impaired in the spleens of WAS-KO mice. Thus, was gene deletion may lead to anemia and hemolysis-related disease, mainly due to increased osmotic fragility of red blood cells, low hemoglobin, increased bilirubin levels and increased serum ferritin. These results indicate that senescence and apoptosis of blood cells also play an important role in the occurrence of WAS. However, most studies have focused only on the immune response. Therefore, the interesting findings of this paper can provide a stronger theoretical basis for further study and help improve the treatment of WAS. We performed gene expression profiling analysis using data obtained from RNA-seq of 2 different group of WT and WAS-KO mice.

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Foamy virus vector-mediated gene correction of a mouse model of Wiskott-Aldrich syndrome.

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Foamy virus vector-mediated gene correction of a mouse model of Wiskott-Aldrich syndrome.

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