Evaluation of Italian Patients with Leber Congenital Amaurosis due to AIPL1 Mutations Highlights the Potential Applicability of Gene Therapy
Evaluation of Italian Patients with Leber Congenital Amaurosis due to AIPL1 Mutations Highlights the Potential Applicability of Gene Therapy
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DOI：
10.1167/iovs.10-6543
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Simonelli, Francesca
中科院分区：
文献类型：
--
作者：
Testa, Francesco;Surace, Enrico Maria;Simonelli, Francesca
目的。目的评价基于基因传递的方法治疗AIPL1基因突变所致Leber先天性黑色素4(LCA4)的可行性。方法：应用基因芯片和直接测序技术分析患者的基因组DNA。对AIPL1基因突变患者进行详细的临床评估，包括眼底自发荧光(Faf)和光学相干断层扫描(OCT)。AIPL1基因缺失的小鼠和猪眼视网膜下注射携带人AIPL1编码序列的腺相关病毒(AAV)载体。P.W278X序列变异是最常见的一个。临床评估揭示了包括弥漫性视网膜营养不良和黄斑病变在内的共同特征。然而，光学相干断层扫描显示，所有年龄段的黄斑外区域的光感受器都有部分保留。眼底自发荧光在后极部可见，而在中心凹内无。AAV介导的Aipl1-/-小鼠的基因转移与AIPL1和βPDE在光感受器上的表达恢复和防止退化有关。临床相关剂量的AAV2/8-AIPL1应用于临床前的大型猪视网膜，可在无毒性的情况下导致AIPL1高水平的光感受器表达。结论：利用先进的影像诊断技术，我们发现黄斑病变是LCA4的主要特征。我们确定了视网膜后极的区域，即使在成年LCA4患者中也存在存活的光感受器，这可能是基因治疗的目标。此外，在Aipl 1-/-小鼠中观察到的对光感受器退化的保护，以及在AAV2/8将AAV2/8传递到大的猪视网膜后观察到的高水平的光感受器转导，支持了对LCA4的可能的基因治疗。(投资眼科VS科学。2011年；52：56185624)doi：10.1167/iovs.10-6543
PURPOSE. To evaluate the suitability of gene delivery-based approaches as potential treatment of Leber congenital amaurosis 4 (LCA4) due to AIPL1 mutations.METHODS. Genomic DNA from patients was analyzed using a microarray chip and direct sequencing. A detailed clinical evaluation including fundus autofluorescence (FAF) and optical coherence tomography (OCT) was performed in patients with AIPL1 mutations. Aipl1 null mice and porcine eyes were subretinally injected with adeno-associated viral (AAV) vectors harboring the human AIPL1 coding sequence.RESULTS. We identified 10 LCA4 patients with mutations in AIPL1. The p. W278X sequence variation was the one most frequently found. Clinical assessment revealed common features including diffuse retinal dystrophies and maculopathy. However, optical coherence tomography showed partially retained photoreceptors in extramacular regions at all ages. The fundus autofluorescence was elicitable at the posterior pole and absent in the fovea. AAV-mediated gene transfer in Aipl1 -/- mice was associated with restoration of AIPL1 and beta PDE expression in photoreceptors and protection from de-generation. Administration of a clinically relevant dose of AAV2/8-AIPL1 to the preclinical large porcine retina resulted in high level of AIPL1 photoreceptor expression in the absence of toxicity.CONCLUSIONS. Using advanced imaging diagnostics we showed that maculopathy is a main feature of LCA4. We identified retinal areas at the posterior pole with surviving photoreceptors present even in adult LCA4 patients, which could be the target of gene therapy. The possible use of gene therapy for LCA4 is additionally supported by the protection from photoreceptor degeneration observed in Aipl 1-/- mice and by the high levels of photoreceptor transduction in the absence of toxicity observed after AAV2/8 delivery to the large porcine retina. (Invest Ophthalmol Vis Sci. 2011;52:5618-5624) DOI:10.1167/iovs.10-6543