Accelerating vision restoration with in-vivo cellular imaging of retinal function

 描述(由申请人提供)：缺乏对活体眼睛的视网膜回路进行成像的方法是所有视力恢复努力的根本障碍。同时评估体内多个视网膜层视网膜回路的结构和功能的能力不仅将改变我们对不同视力恢复策略成败的理解，这些工具将提供的反馈将极大地缩短所有采用它们的方法的开发周期。罗切斯特大学高级视网膜成像联盟(ARIA)的研究人员(詹妮弗·亨特、比尔·梅里根和大卫·威廉姆斯)将创建一种新的视网膜成像工具，旨在跟踪单个动物随着时间的推移，在细胞分辨率下内视网膜和外视网膜结构和功能的变化。该工具将结合遗传编码的钙指示剂的双光子、自适应光学成像来跟踪两种不同动物模型--小鼠和猴子--中单个光感受器和神经节细胞的神经元反应。我们将评估这种成像技术在三种不同的视力恢复方法中的价值。将这些工具应用于基因治疗方法，罗切斯特团队将与康妮·切普科在哈佛大学的实验室合作，通过上调抗氧化剂的表达来挽救视网膜色素变性的视锥功能。我们将与Miescher生物医学研究所的Botond Roska合作，通过在猴子视锥细胞中表达通道视紫红质，来展示这些成像工具对于光遗传学方法的价值。这项工作还将开发一种视网膜变性的灵长类动物模型，在这种模型中，病毒载体被用来刮除光感受器外部节段。最后，这些成像工具将与威斯康星大学麦迪逊分校的David Gamm合作，用于监测小鼠和猴子体内干细胞的分化和神经连接。该联盟不仅将开发先进的成像技术，还将把它转化为视力恢复领域，并创造一个分享最佳实践和结合不同方法的协作环境。

 DESCRIPTION (provided by applicant): The lack of methods to image functioning retinal circuitry in the living eye is a fundamental impediment to all efforts at vision restoration. The ability to assess the structure and function of retinal circuitry in vivo at multiple retinal layer simultaneously will not only transform our understanding of where different vision restoration strategies succeed and fail, the feedback these tools will provide will greatly shorten the development cycle for all approaches in which they are deployed. Investigators in the Advanced Retinal Imaging Alliance (ARIA) at the University of Rochester (Jennifer Hunter, Bill Merigan, and David Williams) will create a new retinal imaging tool designed to track changes in structure and function in both the inner and outer retina at cellular resolution in individual animals over time. This tool will combine two-photon, adaptive optics imaging of genetically-encoded calcium indicators to track the neuronal responses of individual photoreceptors and ganglion cells in two different animal models, mouse and monkey. We will assess the value of this imaging technology in three different approaches to vision restoration. Applying these tools to a gene therapy approach, the Rochester team will collaborate with Connie Cepko's laboratory at Harvard University to rescue cone function in retinitis pigmentosa through the up-regulation of anti-oxidants. We will demonstrate the value of these imaging tools for an optogenetics approach by expressing channel-rhodopsin in monkey cones in collaboration with Botond Roska at the Miescher Institute for Biomedical Research. This effort will also develop a primate model of retinal degeneration in which a viral vector is used to shave off photoreceptor outer segments. Finally, these imaging tools will be deployed to monitor the differentiation and neural connectivity of stem cells in mouse and then monkey in collaboration with David Gamm at the University of Wisconsin, Madison. This consortium will not only develop advanced imaging technology, it will translate it to the field of vision restoration, and create a collaborative environment for sharing best practices and combining different approaches.