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

Cyp21a1 的肾上腺外表达用于先天性肾上腺增生症的基因治疗

• 批准号: 22591148
• 负责人: NAIKI Yasuhiro
• 金额: $ 2.83万
• 依托单位: National Research Institute for Child Health and Development
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22591148/
• 关键词: 遺伝子治療; 副腎皮質過形成; レトロウィルスベクター; アデノウィルス関連ウィルスベクター; 先天性副腎皮質過形成; ips細胞; 21水酸化酵素欠損症; 線維芽細胞; 小児内分泌; ウィルスベクター

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.

21-羟化酶缺乏（CYP21A1D）小鼠是通过用H-2AW18单倍型小鼠的杂合繁殖而成的。构建了含有小鼠CYP21A1 cDNA的逆转录病毒载体，并将其传播到从21OHD小鼠中培养的成纤维细胞。将CYP21A1诱导的成纤维细胞注射到CYP21A1D小鼠中。用PAAV-CMV-Shuttle构建了含有CYP21A cDNA的腺体相关病毒载体（AAV-CYP21A1），并注入了21OHD小鼠的四肢肌肉中。还测量了在注射前后的血清P4和DOC浓度。将转基因成纤维细胞移植到CYP21A1D小鼠中显示出轻微的变化血清P4/DOC比。注射后4周，具有AAV-CYP21A1的转基因21OHD小鼠在4周时显示出DOC的产生增加，P4/DOC比（1223至39.2）降低。我们通过将CYP21A1基因引入肌肉中的肌肉中，在CYP21A1D小鼠中成功恢复了21OH活性类固醇的产生4周。

项目成果

Novel intronic CYP21A2 mutation in Japanese patient with classic salt-wasting steroid 21-hydroxylase deficiency

日本典型耗盐类固醇 21-羟化酶缺乏症患者的新型内含子 CYP21A2 突变

• 发表时间: 2010
• 期刊: Metabolism
• 影响因子: 9.8
• 作者: Nagamitsu S;et al;Noriyuki Katsumata
• 通讯作者: Noriyuki Katsumata