Successful therapeutic effect in a mouse model of erythropoietic protoporphyria by partial genetic correction and fluorescence-based selection of hematopoietic cells
Successful therapeutic effect in a mouse model of erythropoietic protoporphyria by partial genetic correction and fluorescence-based selection of hematopoietic cells
复制标题
通过部分基因校正和基于荧光的造血细胞选择，在红细胞生成性原卟啉症小鼠模型中取得了成功的治疗效果
作者：
A. Fontanellas;M. Méndez;F. Mazurier;M. Cario;S. Navarro;C. Ged;L. Taine;F. Géronimi;E. Richard;F. Moreau;R. Salamanca;H. Verneuil
红细胞生成性原卟啉症的临床特征是皮肤光敏性，生化特征是铁螯合酶缺乏，导致光反应性原卟啉在红细胞、血浆和其他器官中过度积累。Fechm 1 Pas/Fechm 1 Pas小鼠模型的可用性使我们能够测试基因治疗方案，以纠正卟啉表型。基因治疗通过将人铁螯合酶cDNA与逆转录病毒载体离体转移至缺陷造血细胞，然后在卟啉小鼠中重新注射有或无选择的转导细胞来进行。通过FACS将遗传校正的细胞与缺陷的细胞分离，因为当在紫外光下照射时不存在荧光。移植后5个月，荧光红细胞的数量从61%（EPP小鼠）下降到19%的EPP小鼠移植低荧光选择的BM细胞。在荧光RBC少于20%的小鼠中观察到皮肤光敏性缺失。部分表型校正被发现的动物与20至40%的荧光红细胞。总之，骨髓细胞的部分校正足以逆转卟啉表型并恢复正常的造血。这种选择系统代表了一种快速有效的方法，是在逆转录病毒载体中使用潜在有害基因标记的一种极好的替代方法。
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.
原卟啉症中血红素的合成。
DOI：
--
发表时间：
1991
期刊：
Current problems in dermatology
影响因子：
--
作者：
Bloomer,JR;Hill,HD;Kools,AM;Straka,JG
通讯作者：
Straka,JG
影响因子：
2.7
作者：
Miller Ad;Rosman Gj
通讯作者：
Miller Ad;Rosman Gj
DOI：
10.1046/j.1523-1747.1999.00637.x
发表时间：
1999-07
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
X. Wang;L. Yang;L. Kurtz;A. Lichtin;V. Deleo;J. Bloomer;M. Poh-Fitzpatrick
通讯作者：
X. Wang;L. Yang;L. Kurtz;A. Lichtin;V. Deleo;J. Bloomer;M. Poh-Fitzpatrick