Correction of the disease phenotype in the mouse model of Stargardt disease by lentiviral gene therapy
Correction of the disease phenotype in the mouse model of Stargardt disease by lentiviral gene therapy
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DOI：
10.1038/gt.2008.78
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发表时间：
2008-10-01
期刊：
影响因子：
5.1
通讯作者：
Allikmets, R.
中科院分区：
文献类型：
--
作者：
Kong, J.;Kim, S-R;Allikmets, R.
常染色体隐性遗传性Stargardt病（STGD 1）是由ABCA 4（ABCR）基因突变引起的黄斑营养不良。在STGD 1患者以及Abca 4（/）小鼠（一种疾病模型）中最常见的疾病表型是视网膜色素上皮中的脂褐素积累。在这里，我们测试了通过慢病毒载体将正常（wt）人ABCA 4基因递送到Abca 4（/）小鼠的视网膜下腔是否会纠正疾病表型;即，减少脂褐素色素A2 E的积累。构建马传染性贫血病毒（EIAV）衍生的慢病毒载体，其在组成型（CMV）或光感受器特异性（Rho）启动子的控制下表达人ABCA 4基因或LacZ报告基因。在出生后第4天和第5天，在Abca 4（/）小鼠的一只眼睛中视网膜下注射1 μ l（类似于5.0 × 105 TU）的每种EIAV载体。注射生理盐水、EIAV无效载体或未注射的对侧眼作为对照。在注射后的不同时间处死小鼠以测定光感受器（PR）转导效率和A2 E浓度。EIAV-LacZ载体转导小鼠注射区域中5 - 20%的PR。最重要的是，与未处理的和模拟处理的对照眼相比，将EIAV-CMV-ABCA 4单次视网膜下注射至Abca 4（i）小鼠眼实质上减少了疾病相关的A2 E积累。Abca 4（/）小鼠的治疗眼每眼累积8-12 pmol（s. D.= 2.7)治疗后1年，A2 E的量与wt对照相当，而模拟治疗或未治疗的眼睛具有3-5倍的A2 E（每只眼睛27-39 pmol，s. D. = 1.5; P = 0.001-0.005）。尽管外推到人类需要谨慎，但杆和锥光感受器的高转导效率以及STGD 1小鼠模型中A2 E积累的统计学显著减少表明慢病毒基因疗法是治疗ABCA 4相关疾病的潜在有效工具。
Autosomal recessive Stargardt disease (STGD1) is a macular dystrophy caused by mutations in the ABCA4 (ABCR) gene. The disease phenotype that is most recognized in STGD1 patients, and also in the Abca4(/) mouse (a disease model), is lipofuscin accumulation in retinal pigment epithelium. Here, we tested whether delivery of the normal (wt) human ABCA4 gene to the subretinal space of the Abca4(/) mice via lentiviral vectors would correct the disease phenotype; that is, reduce accumulation of the lipofuscin pigment A2E. Equine infectious anemia virus (EIAV)-derived lentiviral vectors were constructed expressing either the human ABCA4 gene or the LacZ reporter gene under the control of the constitutive (CMV) or photoreceptor-specific (Rho) promoters. Abca4(/) mice were injected subretinally with 1 mu l (similar to 5.0 X 105 TU) of each EIAV vector in one eye at postnatal days 4 and 5. An injection of saline, an EIAV-null vector, or an uninjected contralateral eye served as a control. Mice were killed at various times after injection to determine photoreceptor (PR) transduction efficiency and A2E concentrations. EIAV-LacZ vectors transduced from 5 to 20% of the PRs in the injected area in mice. Most importantly, a single subretinal injection of EIAV-CMV-ABCA4 to Abca4(/) mouse eyes substantially reduced disease-associated A2E accumulation compared to untreated and mock-treated control eyes. Treated eyes of Abca4(/) mice accumulated 8-12 pmol per eye (s. d.= 2.7) of A2E 1 year after treatment, amounts comparable to wt controls, whereas mock-treated or untreated eyes had 3-5 times more A2E (27-39 pmol per eye, s. d. = 1.5; P = 0.001-0.005). Although extrapolation to humans requires caution, the high transduction efficiency of both rod and cone photoreceptors and the statistically significant reduction of A2E accumulation in the mouse model of STGD1 suggest that lentiviral gene therapy is a potentially efficient tool for treating ABCA4-associated diseases.