Modeling of congenital erythropoietic porphyria by RNA interference: a new tool for preclinical gene therapy evaluation
✍ Scribed by Elodie Robert-Richard; Magalie Lalanne; Isabelle Lamrissi-Garcia; Véronique Guyonnet-Duperat; Emmanuel Richard; Vincent Pitard; Fréderic Mazurier; François Moreau-Gaudry; Cécile Ged; Hubert de Verneuil
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- English
- Weight
- 357 KB
- Volume
- 12
- Category
- Article
- ISSN
- 1099-498X
- DOI
- 10.1002/jgm.1478
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
Background
Congenital erythropoietic porphyria (CEP) is a severe autosomal recessive disorder characterized by a deficiency in uroporphyrinogen III synthase (UROS), the fourth enzyme of the heme biosynthetic pathway. We recently demonstrated the definitive cure of a murine model of CEP by lentiviral vector‐mediated hematopoietic stem cell (HSC) gene therapy. In the perspective of a gene therapy clinical trial, human cellular models are required to evaluate the therapeutic potential of lentiviral vectors in UROS‐deficient cells. However, the rare incidence of the disease makes difficult the availability of HSCs derived from patients.
Methods
RNA interference (RNAi) has been used to develop a new human model of the disease from normal cord blood HSCs. Lentivectors were developed for this purpose.
Results
We were able to down‐regulate the level of human UROS in human cell lines and primary hematopoietic cells. A 97% reduction of UROS activity led to spontaneous uroporphyrin accumulation in human erythroid bone marrow cells of transplanted immune‐deficient mice, recapitulating the phenotype of cells derived from patients. A strong RNAi‐induced UROS inhibition allowed us to test the efficiency of different lentiviral vectors with the aim of selecting a safer vector. Restoration of UROS activity in these small hairpin RNA‐transduced CD34^+^ cord blood cells by therapeutic lentivectors led to a partial correction of the phenotype in vivo.
Conclusions
The RNAi strategy is an interesting new tool for preclinical gene therapy evaluation. Copyright © 2010 John Wiley & Sons, Ltd.