Ye Guo-jie, Budzynski Ewa, Sonnentag Peter, Nork T Michael, Miller Paul E, McPherson Leslie, Ver Hoeve James N, Smith Leia M, Arndt Tara, Mandapati Savitri, Robinson Paulette M, Calcedo Roberto, Knop David R, Hauswirth William W, Chulay Jeffrey D
1 Applied Genetic Technologies Corporation (AGTC) , Alachua, Florida.
2 Covance Laboratories Inc. , Madison, Wisconsin.
Hum Gene Ther Clin Dev. 2016 Mar;27(1):27-36. doi: 10.1089/humc.2015.163.
Applied Genetic Technologies Corporation (AGTC) is developing rAAV2tYF-PR1.7-hCNGB3, a recombinant adeno-associated virus (rAAV) vector expressing the human CNGB3 gene, for treatment of achromatopsia, an inherited retinal disorder characterized by markedly reduced visual acuity, extreme light sensitivity, and absence of color discrimination. We report here results of a study evaluating safety and biodistribution of rAAV2tYF-PR1.7-hCNGB3 in CNGB3-deficient mice. Three groups of animals (n = 35 males and 35 females per group) received a subretinal injection in one eye of 1 μl containing either vehicle or rAAV2tYF-PR1.7-hCNGB3 at one of two dose concentrations (1 × 10(12) or 4.2 × 10(12) vg/ml) and were euthanized 4 or 13 weeks later. There were no test-article-related changes in clinical observations, body weights, food consumption, ocular examinations, clinical pathology parameters, organ weights, or macroscopic observations at necropsy. Cone-mediated electroretinography (ERG) responses were detected after vector administration in the treated eyes in 90% of animals in the higher dose group and 31% of animals in the lower dose group. Rod-mediated ERG responses were reduced in the treated eye for all groups, with the greatest reduction in males given the higher dose of vector, but returned to normal by the end of the study. Microscopic pathology results demonstrated minimal mononuclear cell infiltrates in the retina and vitreous of some animals at the interim euthanasia and in the vitreous of some animals at the terminal euthanasia. Serum anti-AAV antibodies developed in most vector-injected animals. No animals developed antibodies to hCNGB3. Biodistribution studies demonstrated high levels of vector DNA in vector-injected eyes but little or no vector DNA in nonocular tissue. These results support the use of rAAV2tYF-PR1.7-hCNGB3 in clinical studies in patients with achromatopsia caused by CNGB3 mutations.
应用基因技术公司（AGTC）正在研发rAAV2tYF-PR1.7-hCNGB3，这是一种表达人CNGB3基因的重组腺相关病毒（rAAV）载体，用于治疗色盲症，这是一种遗传性视网膜疾病，其特征为视力显著下降、对光极度敏感且无法辨别颜色。我们在此报告一项评估rAAV2tYF-PR1.7-hCNGB3在CNGB3缺陷小鼠中的安全性和生物分布的研究结果。三组动物（每组35只雄性和35只雌性）在一只眼睛接受视网膜下注射1μl含有赋形剂或两种剂量浓度之一（1×10¹²或4.2×10¹²vg/ml）的rAAV2tYF-PR1.7-hCNGB3，并在4周或13周后实施安乐死。在临床观察、体重、食物消耗、眼部检查、临床病理参数、器官重量或尸检时的宏观观察中，未发现与受试物相关的变化。在高剂量组90%的动物和低剂量组31%的动物中，给药后在治疗眼中检测到锥体介导的视网膜电图（ERG）反应。所有组治疗眼中的杆体介导的ERG反应均降低，给予高剂量载体的雄性动物降低最为明显，但在研究结束时恢复正常。显微镜病理结果显示，在中期安乐死时，部分动物的视网膜和玻璃体中有少量单核细胞浸润，在末期安乐死时，部分动物的玻璃体中有少量单核细胞浸润。大多数注射载体的动物产生了血清抗AAV抗体。没有动物产生针对hCNGB3的抗体。生物分布研究表明，注射载体的眼中载体DNA水平很高，但非眼部组织中载体DNA很少或没有。这些结果支持将rAAV2tYF-PR1.7-hCNGB3用于由CNGB3突变引起的色盲症患者的临床研究。
