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Successful therapeutic effect in a mouse model of erythropoietic protoporphyria by partial genetic correction and fluorescence-based selection of hematopoietic cells

Gene Therapy volume 8, pages 618–626 (2001)Cite this article

Abstract

Erythropoietic protoporphyria is characterized clinically by skin photosensitivity and biochemically by a ferrochelatase deficiency resulting in an excessive accumulation of photoreactive protoporphyrin in erythrocytes, plasma and other organs. The availability of the Fechm1Pas/Fechm1Pas murine model allowed us to test a gene therapy protocol to correct the porphyric phenotype. Gene therapy was performed by ex vivo transfer of human ferrochelatase cDNA with a retroviral vector to deficient hematopoietic cells, followed by re-injection of the transduced cells with or without selection in the porphyric mouse. Genetically corrected cells were separated by FACS from deficient ones by the absence of fluorescence when illuminated under ultraviolet light. Five months after transplantation, the number of fluorescent erythrocytes decreased from 61% (EPP mice) to 19% for EPP mice engrafted with low fluorescent selected BM cells. Absence of skin photosensitivity was observed in mice with less than 20% of fluorescent RBC. A partial phenotypic correction was found for animals with 20 to 40% of fluorescent RBC. In conclusion, a partial correction of bone marrow cells is sufficient to reverse the porphyric phenotype and restore normal hematopoiesis. This selection system represents a rapid and efficient procedure and an excellent alternative to the use of potentially harmful gene markers in retroviral vectors.

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Acknowledgements

We are grateful to M Landry for liver histological analysis, to F Belloc for flow cytometry analysis, to I Lamrissi-Garcia, S Landriau and M Sanchez for technical assistance and to JY Daniel and P Costet for the animal facilities. This work was supported by the Institut National de la Santé et de la Recherche Médicale (INSERM No. 9508), by Association Française contre les Myopathies (AFM), by the Spanish Fondo de Investigaciones Sanitarias (FIS No. 00/0446) and Comunidad Autónoma de Madrid (CAM No. 08.6/0003/1999.1). The animal facility was financed by a grant from the Comité Départemental des Pyrénées Atlantiques de Ligue Nationale contre le Cancer, and from Région Aquitaine. A Fontanellas was supported by the Spanish Instituto de Salud Carlos III (ISCIII No. 98/3165).

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Authors and Affiliations

  1. Laboratoire de Pathologie Moléculaire et Thérapie Génique, Université Victor Segalen Bordeaux 2, France

    A Fontanellas, M Mendez, F Mazurier, M Cario-André, C Ged, L Taine, F Géronimi, E Richard, F Moreau-Gaudry & H de Verneuil

  2. Centro de Investigación, Hospital 12 de Octubre, Madrid, Spain

    A Fontanellas, S Navarro & R Enriquez de Salamanca

  3. Laboratoire de Génétique, CHU Pellegrin, Bordeaux, France

    L Taine

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Fontanellas, A., Mendez, M., Mazurier, F. et al. Successful therapeutic effect in a mouse model of erythropoietic protoporphyria by partial genetic correction and fluorescence-based selection of hematopoietic cells. Gene Ther 8, 618–626 (2001). https://doi.org/10.1038/sj.gt.3301427

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