title: Safe and Effective Gene Therapy for Murine Wiskott-Aldrich Syndrome Using an Insulated Lentiviral Vector.
abstract: Wiskott-Aldrich syndrome (WAS) is a life-threatening immunodeficiency caused by mutations within theWASgene. Viral gene therapy to restore WAS protein (WASp) expression in hematopoietic cells of patients with WAS has the potential to improve outcomes relative to the current standard of care, allogeneic bone marrow transplantation. However, the development of viral vectors that are both safe and effective has been problematic. While use of viral transcriptional promoters may increase the risk of insertional mutagenesis, cellular promoters may not achieve WASp expression levels necessary for optimal therapeutic effect. Here we evaluate a self-inactivating (SIN) lentiviral vector combining a chromatin insulator upstream of a viral MND (MPSV LTR, NCR deleted, dl587 PBS) promoter driving WASp expression. Used as a gene therapeutic inWas-/-mice, this vector resulted in stable WASp+cells in all hematopoietic lineages and rescue of T and B cell defects with a low number of viral integrations per cell, without evidence of insertional mutagenesis in serial bone marrow transplants. In a gene transfer experiment in non-human primates, the insulated MND promoter (driving GFP expression) demonstrated long-term polyclonal engraftment of GFP+cells. These observations demonstrate that the insulated MND promoter safely and efficiently reconstitutes clinically effective WASp expression and should be considered for future WAS therapy.
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Swati
authors.initials: S
authors.lastname: Singh
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Iram
authors.initials: I
authors.lastname: Khan
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Socheath
authors.initials: S
authors.lastname: Khim
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Brenda
authors.initials: B
authors.lastname: Seymour
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Sommer
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Wielgosz
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
authors.forename: Zachary
authors.initials: Z
authors.lastname: Norgaard
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Department of Pathology, University of Washington, Seattle, WA 98105, USA.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Kiem
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Department of Medical Oncology, University of Washington, Seattle, WA 98105, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Adair
authors.affiliation: Department of Comparative Medicine, University of Washington, Seattle, WA 98105, USA.
authors.forename: Denny
authors.initials: D
authors.lastname: Liggitt
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
authors.forename: Arthur
authors.initials: A
authors.lastname: Nienhuis
authors.affiliation: Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, WA 98101, USA; Department of Pediatrics, University of Washington, Seattle, WA 98105, USA; Department of Immunology, University of Washington, Seattle, WA 98105, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rawlings
keywords: Wiskott-Aldrich Syndrome; gene therapy; insulated lentivirus