Extra-adrenal expression of Cyp21a1 for gene therapy of congenital adrenal hyperplasia

21-通过与H-2aw 18单倍型小鼠的杂合子交配制备羟化酶缺陷型（Cyp 21 a1 D）小鼠。构建了含Cyp 21 a1基因的逆转录病毒载体，并将其转染21 OHD小鼠成纤维细胞。将Cyp 21 a1诱导的成纤维细胞注射到Cyp 21 a1 D小鼠中。用腺相关病毒载体pAAV-CMV-shuttle构建了Cyp 21 acDNA的腺相关病毒载体（AAV-Cyp 21 a1）。注射前和注射后测定血清P4和DOC浓度。将转基因成纤维细胞移植到Cyp 21 a1 D小鼠体内，血清P4/DOC比值变化不大。AAV-Cyp 21 a1转基因21 OHD小鼠在注射后4周表现出DOC产量增加和P4/DOC比值降低（1223至39.2）。通过腺相关病毒载体将Cyp 21 a1基因导入Cyp 21 a1 D小鼠肌肉中4周，成功地恢复了Cyp 21 a1 D小鼠21 OH活性类固醇产生。

21-hydorxylase deficient (Cyp21a1D) mice were made by breeding with heterozygous of H-2aw18 haplotype mice. Retrovirus vector containing mouse Cyp21a1 cDNA was constructed and transmitted to fibroblasts cultured from 21OHD mice. Cyp21a1-induced fibroblasts were injected into Cyp21a1D mice. Adenoassociated virus vector containing Cyp21a cDNA (AAV-Cyp21a1) was constructed with pAAV-CMV-shuttle and injected into limbs muscles of 21OHD mice. Serum P4 and DOC concentrations were also measured before and after injection. Transplantation of the transgenic fibroblasts into Cyp21a1D mice showed slight change serum P4/DOC ratio. Transgenic 21OHD mice with AAV-Cyp21a1 showed increase DOC production and decreased P4/DOC ratio (1223 to 39.2) at 4weeks after injection. We succeeded recovering 21OH activity steroid production in in Cyp21a1D mice by introducing Cyp21a1 gene into muscle with adenoassociated virus vectors for 4 weeks.