Toxicology and Pharmacology of an AAV Vector Expressing Codon-Optimized RPGR in RPGR-Deficient Rd9 Mice.-猫眼课题宝

DOI：

10.1089/humc.2018.168

发表时间：

2018-12

期刊：

Human gene therapy. Clinical development

影响因子：

通讯作者：

中科院分区：

其他

文献类型：

作者：

Chunjuan Song;T. Conlon;W. Deng;K. Coleman;Ping Zhu;Cayrn Plummer;Savitri Mandapati;Mailin Van Hoosear;Kari B. Green;P. Sonnentag;Alok K. Sharma;A. Timmers;Paulette M. Robinson;D. R. Knop;W. Hauswirth;J. Chulay;M. Shearman;G. Ye

应用遗传技术公司（AGTC）正在开发一种重组腺相关病毒（rAAV）载体AGTC-501，也称为AAV 2 tYF-GRK 1-RPGRco，用于治疗视网膜色素变性（RP）患者的视网膜色素变性（RP）视网膜色素变性磷酸酶调节因子（RPGR）基因突变。该载体含有由光受体特异性启动子（G蛋白偶联受体激酶1，GRK 1）驱动的密码子优化的人RPGR cDNA（RPGRco），并包装在具有三个表面酪氨酸残基改变为苯丙氨酸的AAV 2衣壳（AAV 2 tYF）中。我们在天然存在的RPGR缺陷型Rd 9小鼠模型中进行了通过视网膜下注射施用该载体的安全性和效力研究。六十只Rd 9小鼠（每组20只）在右眼中接受视网膜下注射媒介物（对照）或两种剂量水平（4 × 108或4 × 109 vg/眼）之一的AAV 2 tYF-GRK 1-RPGRco，并在注射后随访12周。载体注射耐受性良好，无全身毒性。高向量剂量组中存在视网膜电图b波振幅降低的趋势，但无统计学显著性。血液学或临床化学参数无临床重要变化，并且在生活中或通过组织学检查无载体相关的眼部变化。剂量依赖性RPGR蛋白表达，主要在光感受器的内段和相邻的连接纤毛区域，观察到在所有载体处理的眼睛检查。通过PCR扩增的DNA或从载体处理的视网膜组织提取的总RNA逆转录的cDNA的测序，以及通过从转染的HEK 293细胞获得的RPGR蛋白的测序，证实密码子优化的RPGR的序列完整性。这些结果支持rAAV 2 tYF-GRK 1-RPGRco用于RPGR突变引起的XLRP患者的临床研究。

Applied Genetic Technologies Corporation (AGTC) is developing a recombinant adeno-associated virus (rAAV) vector AGTC-501, also designated AAV2tYF-GRK1-RPGRco, to treat retinitis pigmentosa (RP) in patients with mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene. The vector contains a codon-optimized human RPGR cDNA (RPGRco) driven by a photoreceptor-specific promoter (G protein-coupled receptor kinase 1, GRK1) and is packaged in an AAV2 capsid with three surface tyrosine residues changed to phenylalanine (AAV2tYF). We conducted a safety and potency study of this vector administered by subretinal a injection in the naturally occurring RPGR-deficient Rd9 mouse model. Sixty Rd9 mice (20 per group) received a subretinal injection in the right eye of vehicle (control) or AAV2tYF-GRK1-RPGRco at one of two dose levels (4 × 108 or 4 × 109 vg/eye) and were followed for 12 weeks after injection. Vector injections were well tolerated, with no systemic toxicity. There was a trend towards reduced electroretinography b-wave amplitudes in the high vector dose group that was not statistically significant. There were no clinically important changes in hematology or clinical chemistry parameters and no vector-related ocular changes in life or by histological examination. Dose-dependent RPGR protein expression, mainly in the inner segment of photoreceptors and the adjacent connecting cilium region, was observed in all vector-treated eyes examined. Sequence integrity of the codon-optimized RPGR was confirmed by sequencing of PCR-amplified DNA, or cDNA reverse transcribed from total RNA extracted from vector-treated retinal tissues, and by sequencing of RPGR protein obtained from transfected HEK 293 cells. These results support the use of rAAV2tYF-GRK1-RPGRco in clinical studies in patients with XLRP caused by RPGR mutations.

Toxicology and Pharmacology of an AAV Vector Expressing Codon-Optimized RPGR in RPGR-Deficient Rd9 Mice.

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