Géronimi F, Richard E, Lamrissi-Garcia I, Lalanne M, Ged C, Redonnet-Vernhet I, Moreau-Gaudry F, de Verneuil H
INSERM E 0217, Université Victor Segalen, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France.
J Mol Med (Berl). 2003 May;81(5):310-20. doi: 10.1007/s00109-003-0438-7. Epub 2003 Apr 30.
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.
先天性红细胞生成性卟啉病（CEP）是一种由于血红素生物合成途径的第四个酶——尿卟啉原III合酶缺乏而导致的遗传性疾病。其特征是尿卟啉I在骨髓、外周血和其他器官中蓄积。CEP的预后很差，常在成年早期死亡。对于严重依赖输血的病例，当无法进行异基因细胞移植时，基因修饰的原始/干细胞自体移植可能是唯一的选择。体外基因转移实验已证明通过造血细胞进行基因治疗来治疗这种疾病的可行性。在本研究中，用治疗性人尿卟啉原III合酶（UROS）cDNA对卟啉细胞系和原代CD34(+)细胞进行慢病毒转导，导致了酶学和代谢方面的纠正，这通过转导细胞中UROS活性的增加和卟啉蓄积的抑制得以证明。在细胞系和CD34(+)细胞中均实现了非常高的基因转移效率（高达90%），且无需任何筛选。转基因的表达在长期液体培养中保持稳定。此外，在CD34(+)细胞的体外红细胞分化过程中基因表达得以维持。因此，慢病毒载体在未来通过基因治疗CEP患者方面具有广阔前景。
