Journal article
piggyBac-mediated phenotypic correction of factor VIII deficiency
Molecular therapy. Methods & clinical development, Vol.1(C), pp.14042-14042
2014
DOI: 10.1038/mtm.2014.42
PMCID: PMC4362371
PMID: 26015980
Abstract
Hemophilia A, caused by a deficiency in factor VIII (FVIII), is the most severe inherited bleeding disorder. Hemophilia A is an attractive gene therapy candidate because even small increases in FVIII levels will positively alter the phenotype. While several vectors are under investigation, gene addition from an integrated transgene offers the possibility of long term expression. We engineered the DNA transposon-based vector, piggyBac (PB), to carry a codon-optimized B-domain deleted human FVIII cDNA. Evaluation of gene transfer efficiency in FVIII null mice demonstrated that PB containing the FVIII cDNA, delivered via hydrodynamic injection to immunocompetent hemophilia mice, conferred persistent gene expression, attaining mean FVIII activity of approximately 60% with 3/19 developing inhibitors. In addition to efficacious expression, a goal of gene transfer-based therapies is to develop vectors with low toxicity. To assess endoplasmic reticulum stress in hepatocytes stably expressing the transgene, we evaluated levels of ER stress markers via qPCR and found no evidence of cell stress. To evaluate phenotypic correction, a tail clip assay performed at the end of the study revealed reduced blood loss. These data demonstrate that PB can be used to achieve sustained FVIII expression and long-term therapeutic benefit in a mouse model.
Details
- Title: Subtitle
- piggyBac-mediated phenotypic correction of factor VIII deficiency
- Creators
- Janice M Staber - Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USA ; Stead Family Department of Pediatrics, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USAMolly J Pollpeter - Stead Family Department of Pediatrics, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USAAngela Arensdorf - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USAPatrick L Sinn - Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USA ; Stead Family Department of Pediatrics, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USAD Thomas Rutkowski - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USAPaul B McCray Jr - Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USA ; Stead Family Department of Pediatrics, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USA ; Interdisciplinary Graduate Program in Genetics, Carver College of Medicine, University of Iowa , Iowa City, Iowa, USA
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Methods & clinical development, Vol.1(C), pp.14042-14042
- Publisher
- United States
- DOI
- 10.1038/mtm.2014.42
- PMID
- 26015980
- PMCID
- PMC4362371
- ISSN
- 2329-0501
- eISSN
- 2329-0501
- Grant note
- R44 HL081976 / NHLBI NIH HHS K12 HD027748 / NICHD NIH HHS P30 DK054759 / NIDDK NIH HHS
- Language
- English
- Date published
- 2014
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Anatomy and Cell Biology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Hematology/Oncology; Internal Medicine
- Record Identifier
- 9984065839502771
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