Leber's Hereditary Optic Neuropathy: Gene Therapy Clinical Trial

描述（由申请人提供）：在过去的十年中，我们在确定Leber's遗传性视神经病变（LHON）的发病机制和测试治疗方面取得了重大进展。我们成功地在核遗传密码中表达了野生型人NADH泛醌氧化还原酶I亚基4 （ND4）。该蛋白通过线粒体靶向序列被导入线粒体。该基因被包装成下一代酪氨酸-苯丙氨酸修饰的自互补腺相关病毒（AAV），然后注射到啮齿动物的眼睛。flag标记的野生型人ND4在注射后1天在大多数视网膜内神经元中快速检测到，并整合到复合物i中。此外，在啮齿类动物的眼睛中，也注射了导致大多数LHON的突变体Gl 1778A ND4同源物，野生型ND4恢复了有缺陷的ATP合成，抑制了视力丧失，减少了视网膜神经节细胞的凋亡，并阻止了视神经轴突的长期死亡。在离体人眼中注射相应滴度的自互补野生型ND4在大多数视网膜神经节细胞中表达，提示其在我们的LHON患者中也会表达。裸鼠体注射ND4无不良反应，提示该载体可作为LHON临床试验安全有效的平台。我们的目标是在G11778A mtDNA突变患者的I期临床试验中测试aav介导的人类ND4基因递送的安全性，然后进入II期研究，以证明该项目后期的有效性。第一阶段将包括一个非盲、单边,单剂intravitreal注入AAV-ND4每个病人的糟糕眼睛增大剂量研究调查三个向量的安全剂量(2.46 5 x10e9 vg, x10e10 vg和1 xloel1 vg)在少数分子证实Gl 1778线粒体DNA突变患者有慢性双边、严重的视力丧失超过1年(目标1)或急性双边几个视觉损失更少的那1年(目标2),然后,最后,在视力较好的眼睛中，但我们知道，在第一只眼睛视力丧失开始后的6个月内，注定会失去明显的视力（目标3）。

DESCRIPTION (provided by applicant): Over the past decade, we have made major strides towards determining the pathogenesis and testing a treatment for Leber's Hereditary Optic Neuropathy (LHON). We successfully expressed the wild-type human NADH ubiquinone oxidoreductase subunit 4 (ND4) of complex I in the nuclear genetic code. The protein was imported into the mitochondria by agency of a mitochondrial targeting sequence. The gene was packaged into next generation tyrosine to phenylalanine modified self-complementary adenoassociated virus (AAV) then injected into rodent eyes. FLAG-tagged wild-type human ND4 was detected quickly in most inner retinal neurons by 1 day post injection and it integrated into Complex I. Furthermore, in rodent eyes also injected with a mutant Gl 1778A ND4 homologue responsible for most cases of LHON, wild-type ND4 restored defective ATP synthesis, suppressed visual loss, reduced apoptosis of retinal ganglion cells and prevented demise of axons in the optic nerve that persisted long-term. The self-complementary wild-type ND4 injected at the relevant titer into the ex vivo human eye expressed in most retinal ganglion cells, suggesting that it will do so in our LHON patients. Primates vitreally injected with untagge ND4 had no adverse reactions, suggesting that this vector should be a safe and effective platform for clinical testing in LHON. Our goal in this application is to test the safety of AAV-mediated delivery of the human ND4 gene in a Phase I clinical trial of patients with mutated G11778A mtDNA and then move to a Phase II study to prove efficacy in the later years of this program. Phase I will consist of an open-label, unilateral, single-dose intravitreal injection of AAV-ND4 per patient in the worse eye in a dose-escalation study investigating the safety of three vector doses (5x10e9 vg, 2.46x10e10 vg and 1xlOel1 vg) in a small number of patients with molecularly confirmed Gl 1778A-mutated mitochondrial DNA who have chronic bilateral, severe visual loss for more than 1 year (Aim 1) or acute bilateral several visual loss for less tha 1 year (Aim 2), and then, lastly, in the eye with better vision but that we know is predestined to lose significant vision within 6 months from the onset of visual loss in the first eye (Aim 3).