Metabolic correction of congenital erythropoietic porphyria with iPSCs free of reprogramming factors.

Abstract

Congenital erythropoietic porphyria (CEP) is due to a deficiency in the enzymatic activity of uroporphyrinogen III synthase (UROS); such a deficiency leads to porphyrin accumulation and results in skin lesions and hemolytic anemia. CEP is a candidate for retrolentivirus-mediated gene therapy, but recent reports of insertional leukemogenesis underscore the need for safer methods. The discovery of induced pluripotent stem cells (iPSCs) has opened up new horizons in gene therapy because it might overcome the difficulty of obtaining sufficient amounts of autologous hematopoietic stem cells for transplantation and the risk of genotoxicity. In this study, we isolated keratinocytes from a CEP-affected individual and generated iPSCs with two excisable lentiviral vectors. Gene correction of CEP-derived iPSCs was obtained by lentiviral transduction of a therapeutic vector containing UROS cDNA under the control of an erythroid-specific promoter shielded by insulators. One iPSC clone, free of reprogramming genes, was obtained with a single proviral integration of the therapeutic vector in a genomic safe region. Metabolic correction of erythroblasts derived from iPSC clones was demonstrated by the disappearance of fluorocytes. This study reports the feasibility of porphyria gene therapy with the use of iPSCs.

• Authors: Aurélie Bedel, Miguel Taillepierre, Véronique Guyonnet-Duperat, Eric Lippert, Pierre Dubus, Sandrine Dabernat, Thibaud Mautuit, Bruno Cardinaud, Catherine Pain, Benoît Rousseau, Magalie Lalanne, Cécile Ged, Yann Duchartre, Emmanuel Richard, Hubert de Verneuil, François Moreau-Gaudry
• Journal: American journal of human genetics (Am J Hum Genet) Vol. 91 Issue 1 Pg. 109-21 (Jul 13 2012) ISSN: 1537-6605 [Electronic] United States
• PMID: 22795135 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 The American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.

Chemical References

• Uroporphyrinogen III Synthetase

Topics

• Cell Differentiation
• Feasibility Studies
• Genetic Therapy (methods)
• Genetic Vectors
• Hematopoietic Stem Cells (cytology)
• Humans
• Induced Pluripotent Stem Cells (transplantation)
• Keratinocytes (cytology)
• Lentivirus (genetics)
• Porphyria, Erythropoietic (genetics, therapy)
• Transduction, Genetic
• Uroporphyrinogen III Synthetase (genetics)