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Lentivirus-mediated gene transfer of uroporphyrinogen III synthase fully corrects the porphyric phenotype in human cells

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Congenital erythropoietic porphyria (CEP) is an inherited disease due to a deficiency in the uroporphyrinogen III synthase, the fourth enzyme of the heme biosynthesis pathway. It is characterized by accumulation of uroporphyrin I in the bone marrow, peripheral blood and other organs. The prognosis of CEP is poor, with death often occurring early in adult life. For severe transfusion-dependent cases, when allogeneic cell transplantation cannot be performed, the autografting of genetically modified primitive/stem cells may be the only alternative. In vitro gene transfer experiments have documented the feasibility of gene therapy via hematopoietic cells to treat this disease. In the present study lentiviral transduction of porphyric cell lines and primary CD34+ cells with the therapeutic human uroporphyrinogen III synthase (UROS) cDNA resulted in both enzymatic and metabolic correction, as demonstrated by the increase in UROS activity and the suppression of porphyrin accumulation in transduced cells. Very high gene transfer efficiency (up to 90%) was achieved in both cell lines and CD34+ cells without any selection. Expression of the transgene remained stable over long-term liquid culture. Furthermore, gene expression was maintained during in vitro erythroid differentiation of CD34+ cells. Therefore the use of lentiviral vectors is promising for the future treatment of CEP patients by gene therapy.

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Bone marrow transplant


Congenital erythropoietic porphyria


Colony-forming cell


Enhanced green fluorescent protein

Epo :


FL :

FLT3 ligand


Human immunodeficiency virus


Hematopoietic stem/progenitor cell

IL :


LB :



Lymphoblastoid cells from patients with congenital erythropoietic porphyria


Lymphoblastoid cells from normal individuals


Long-term culture initiating cell


Long-term culture medium


Long terminal repeat


Multiplicity of infection

mPB :

Cytokine-mobilized peripheral blood


RM-B00 medium + cytokines


Stem cell factor


Severe combined immunodeficiency










Uroporphyrinogen III synthase


Woodchuck hepatitis virus posttranscriptional regulatory element


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We thank P. Charneau, A. Dubart-Kupperschmidt, and F. Pflumio for donating the TRIPΔU3-EF1α vector, J. Reiffers and B. Dazey for the normal CD34+ mPB cells, and M. Morey for the porphyric cells. This work was supported by grants from Association Française contre les Myopathies and Conseil Régional d'Aquitaine.

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Correspondence to H. de Verneuil.

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Géronimi, F., Richard, E., Lamrissi-Garcia, I. et al. Lentivirus-mediated gene transfer of uroporphyrinogen III synthase fully corrects the porphyric phenotype in human cells. J Mol Med 81, 310–320 (2003).

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