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。我们成功地表达了野生型人类NADH泛醌氧化还原酶亚基4（ND 4）的复合物I的核遗传密码。该蛋白质通过线粒体靶向序列的代理被导入线粒体。该基因被包装到下一代酪氨酸到苯丙氨酸修饰的自身互补腺相关病毒（AAV）中，然后注射到啮齿动物眼中。FLAG标记的野生型人ND 4在注射后1天在大多数内视网膜神经元中快速检测到，并且其整合到复合物I中。此外，在也注射了导致大多数LHON病例的突变型G11778 A ND 4同源物的啮齿动物眼中，野生型ND 4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，并防止了长期持续存在的视神经中轴突的死亡。以相关滴度注射到离体人眼中的自身互补野生型ND 4在大多数视网膜神经节细胞中表达，这表明它将在我们的LHON患者中表达。未标记的ND 4经玻璃体注射灵长类动物无不良反应，提示该载体可作为LHON临床试验的安全有效平台。我们在这项申请中的目标是在G11778 A mtDNA突变患者的I期临床试验中测试AAV介导的人ND 4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疗效。I期将包括在剂量递增研究中，在每名患者的较差眼中开放标签、单侧、单剂量玻璃体内注射AAV-ND 4，研究三种载体剂量的安全性。（5x 10 e9 vg、2.46x10e10vg和1x 10 e11 vg），这些患者具有经分子学证实的G11778 A突变的线粒体DNA，严重视力丧失超过1年（目标1）或急性双侧几个视力丧失少于1年（目标2），然后，最后，视力较好的眼睛，但我们知道是注定要失去显着的视力在6个月内从第一只眼睛视力丧失发作（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).