Optogenetic Vision Restoration

光遗传学视力恢复

• 批准号: 10004654
• 负责人: John Gerard Flannery
• 金额: $ 39.25万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004654
• 关键词: Affect; Age; American; Animals; Automobile Driving; Behavior; Behavioral; Blindness; Brain; Categories; Cell Death; Cells; Characteristics; Chemicals; Choroideremia; Clinic; Complement; Complex; Cone; Cues; Data; Disease; Engineering; Eye; Eye diseases; FDA approved; Formulation; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Glutamate Receptor; Goals; Goggles; Head; Image; Inherited; Injections; Interneurons; Kinetics; Light; Medical; Motion; Mus; Mutation; Natural regeneration; Neurons; Opsin; Outcome; Output; Patients; Pattern; Pattern Recognition; Performance; Photophobia; Photoreceptors; Plug-in; Population; Property; Proteins; Research; Resolution; Retina; Retinal Degeneration; Retinal Diseases; Retinitis Pigmentosa; Risk; Rod; S-nitro-N-acetylpenicillamine; Schedule; Shock; Signal Transduction; Speed; System; Testing; Therapeutic; Time; Vertebrate Photoreceptors; Virus; Vision; Visual; Visual Acuity; Wild Type Mouse; base; combinatorial; design; experimental study; foot; ganglion cell; gene therapy; improved; inherited retinal degeneration; intravitreal injection; light intensity; microbial; novel strategies; object recognition; optical sensor; optogenetics; photoreceptor degeneration; preference; programs; receptor; reconstitution; relating to nervous system; response; restoration; retinal neuron; retinal prosthesis; sensor; success

Over 100,000 Americans of all ages suffer from inherited retinal diseases (IRD), which cause a progressive loss of vision. In most IRDs, disease begins in the rods, causing vision loss from the periphery to the center, leaving patients unable to navigate their surroundings. Electronic retinal prosthesis restore useful vision in patients affected by IRDs, and optogenetics is an alternative therapeutic. A major limitation of microbial opsins for restoration of retinal light sensitivity is the high light intensity required for activating channelrhodopsins. A solution to this caveat is the use of opsins with higher light sensitivity but sufficiently fast kinetics for useful motion vision. We propose a novel approach to restore vision to patients using a virus to express a light sensitive protein in specific, second-order retina neurons to make them light sensitive. Our approach uses a common neuronal receptor, modified to add a light receptive function to the remaining light-insensitive retinal neurons that survive after photoreceptor degeneration. The receptor uses either retinal, which is available in the eye, or a synthetic chemical photoswitch delivered by intravitreal injection. In this way, the cells in which the receptor is located respond to light with a change in neural firing. This compensates for their loss of input from photoreceptors, restoring light responsiveness to the retina and sending information to the brain to restore vision. In most cases, this approach is independent of the mutation that caused the photoreceptor degeneration. Exceptions to this approach may be diseases that cause RPE cell death, such as choroideremia. To date, versions of this approach, developed by Co-PIs Isacoff and Flannery, and others in the field, have employed receptors that are rather insensitive to light or very slow in response and so could not support normal vision. We now propose a new strategy that uses the natural amplification properties of GPCR signaling to increase sensitivity (by 1000 times) and speed. GPCR signaling cascades are intrinsic to rods and cones, as well as bipolar, ganglion cells and other cells in the retina. We also pursue a new discovery, emerging from our preliminary experiments, which enables a combinatorial approach that uses more than one optical sensor molecule at a time in order to recreate the natural diversity of natural signaling in the retina that had earlier been missing. Finally, we employ sophisticated behavioral analysis to test not only the restoration of the ability to tell light from dark or flashing from steady light, but to determine if the animal is able to see images. Success of this program would represent a major step in the creation of a retinal prosthetic based on gene therapy.

所有年龄段的超过100,000名美国人患有遗传性视网膜疾病（IRD），这会导致视力丧失。在大多数IRD中，疾病始于杆子，导致外围的视力丧失到中心，使患者无法在周围环境中度过。电子视网膜假体恢复了受IRD影响的患者的有用视力，而光遗传学是一种替代性治疗。微生物蛋白用于恢复视网膜光敏感性的主要局限性是激活通道旋转所需的高光强度。解决此警告的一种解决方案是使用具有较高光灵敏度的OPSIN，但具有足够快的动力学，以实现有用的运动视觉。 我们提出了一种新型方法，以恢复使用病毒表达光敏病毒的患者的视力 特定的二阶视网膜神经元中的蛋白质使其对光敏。我们的方法使用常见 神经元受体，被修饰以在其余光线视网膜上添加光感受功能 光感受器变性后生存的神经元。该受体使用的是可用的视网膜 在眼中，或通过玻璃体内注射提供的合成化学照片开关。这样，单元格 受体所定位的对光的反应，随着神经发射的变化。这弥补了他们的损失 来自光感受器的输入，恢复对视网膜的光响应能力并将信息发送给 大脑恢复视力。在大多数情况下，这种方法独立于引起的突变 感光体变性。这种方法的例外可能是导致RPE细胞死亡的疾病，这样 作为绒毛膜血症。 迄今 领域，使用的受体对响应的光或非常缓慢不敏感，因此无法 支持正常视力。现在，我们提出了一种使用自然放大属性的新策略 GPCR信号传导以提高灵敏度（提高1000倍）和速度。 GPCR信号级联是固有的 到视网膜中的杆和锥，以及双极，神经节细胞和其他细胞。我们也追求新的 发现，从我们的初步实验中得出 一次以上的光学传感器分子以重新创建自然的自然多样性 先前缺失的视网膜信号。最后，我们采用复杂的行为分析来 不仅可以测试从稳定光中从黑暗或闪烁的灯光来恢复的能力，还可以确定 如果动物能够看到图像。该计划的成功将代表创建的重大步骤 基于基因治疗的视网膜假体。