A Next Generation Outer Retinal Microphysiological System

摘要 诱导多能干细胞(IPSCs)的发现为科学家提供了发展的机会 遗传性疾病患者的分化组织，并使用这些体外模型来研究疾病 并测试药物和基于基因的治疗方法。然而，当前的区分协议经常失败 以准确概括天然组织中存在的细胞组织。例如，即使是最先进的- ART视网膜分化方案，这导致了三维视网膜器质，不准确 重述存在于视网膜外层的细胞与细胞的相互作用。特别是脉络膜内皮细胞 细胞很少出现，并且永远不会在RPE下面形成有组织的层。类似地，与其形成一个 与新形成的光感受器并列的单层RPE细胞通常以簇状分布在 神经视网膜的边缘。在本申请中，我们描述了一个专注于开发和 验证微生理系统的工程设计，以准确地重建复杂的架构 外面的视网膜被灌流。具体地说，我们建议将超高分辨率、最先进的3D打印 视网膜支架、IPSC技术和微流控技术开发下一代微生理系统 它概括了视网膜外部的单位。完成该提案中概述的目标将有助于铺平道路 未来的研究重点是评估复杂的视网膜退行性疾病的病理生理学，如 老年性黄斑变性与新药和基于基因的疗法的发展。

ABSTRACT The discovery of induced pluripotent stem cells (iPSCs) provided scientists with the opportunity to develop differentiated tissues from patients with inherited disease, and to use these in vitro models to study disease mechanisms and to test drug and gene-based therapeutics. However, current differentiation protocols often fail to accurately recapitulate the cellular organization present in the native tissue. For example, even state-of-the- art retinal differentiation protocols, which give rise to three dimensional retinal organoids, do not accurately recapitulate the cell-cell interactions that are present in the outer retina. Specifically, choroidal endothelial cells are rarely present and never form an organized layer beneath the RPE. Similarly, rather than forming a monolayer juxtaposed to newly developed photoreceptors, RPE cells are often found in clusters located at the edges of the neural retina. In this application we describe a program focused on the development and validation of a microphysiological system engineered to accurately recreate the complex architecture of the perfused outer retina. Specifically, we propose to combine ultra-high resolution, state-of-the-art 3D printing of retinal scaffolds, iPSC technology, and microfluidics to develop a next generation microphysiological system that recapitulates the outer retina unit. Completion of the aims outlined in this proposal will help pave the way for future studies focused on evaluating the pathophysiology of complex retinal degenerative disorders such as age-related macular degeneration and development of novel drug and gene-based therapeutics.