Hippo Pathway Regulation of Müller Glial Cell-mediated Retinal Regeneration

项目总结/摘要 视网膜疾病如青光眼、黄斑变性和糖尿病性视网膜病变，以及创伤性视网膜病变， 损伤，导致视网膜神经元丧失，从而丧失视力，剥夺了世界各地许多人最宝贵的感官之一。 因此，迫切需要设计策略来恢复丢失的视网膜神经元，从而导致视力恢复。电流 视网膜再生医学的努力大量投资于细胞替代方法。然而，它也可能 有可能诱导哺乳动物视网膜经历内在的自我修复机制以再生神经元。 已知非哺乳类脊椎动物（例如斑马鱼）的视网膜表现出显著的视网膜电生理能力。 再生在这里，Müller胶质细胞（MG）重新编程为增殖性视网膜祖细胞样细胞， 分化成新的光感受器，导致视力恢复。不幸的是，不知什么原因 哺乳动物MG已经失去了这种能力，或者它处于休眠状态。我们的长期目标是确定细胞和分子 阻断哺乳动物MG介导的视网膜再生的机制。这样我们就能设计出 策略绕过这一系统，从而重新唤醒哺乳动物视网膜的再生潜力。 在这个建议中，我们将测试假设，河马信号通路积极阻断哺乳动物 MG介导的视网膜再生，通过防止持续的MG增殖和重新编程为视网膜细胞， 祖细胞样状态我们的具体目标将精确定义Hippo信号在负调控中的需求 MG对视网膜损伤的持续性细胞周期再进入。我们还将确定是否遗传 绕过Hippo信号传导导致MG重编程为祖细胞样状态。最后，我们将评估 重编程MG的神经原性潜力。 该项目将采用多学科方法，使用遗传功能丧失和获得实验， 命运作图、表观基因组学和单细胞转录组学。通过完成上述目标，我们预计 确定Hippo途径为通常抑制MG增殖的内源性分子机制， 重新编程为可再生的祖细胞这些数据将提供一个重要的分子切入点， 这是为了进一步研究促进Müller神经胶质细胞介导的视网膜再生的新方法， 对视网膜再生医学领域产生重大影响。我们预计后续的调查将涉及更多 本发明涉及用于调节Hippo途径活性以促进视网膜再生的翻译治疗方法。

PROJECT SUMMARY/ABSTRACT Retinal diseases such as glaucoma, macular degeneration, and diabetic retinopathy, as well as traumatic injury, result in loss of retinal neurons and thus sight, depriving many worldwide of one of our most valued senses. Thus, there is a critical need to devise strategies to restore lost retinal neurons leading to vision recovery. Current efforts in retinal regenerative medicine are heavily invested in cell replacement approaches. However, it may also be possible to induce the mammalian retinae to undergo an intrinsic self-repair mechanism to regenerate neurons. The retinae of non-mammalian vertebrates, such as zebrafish, are known to exhibit the remarkable ability of retinal regeneration. Here, Müller glial cells (MGs) reprogram to proliferative, retinal progenitor-like cells that in turn differentiate into new photoreceptors leading to restoration of vision. Unfortunately, for unknown reasons, mammalian MGs have lost this ability, or it is dormant. Our long term goal is to identify the cellular and molecular mechanisms blocking mammalian MG-mediated retinal regeneration. By doing so, we may be able to devise strategies to bypass this system and thereby reawaken the regenerative potential of the mammalian retina. In this proposal, we will test the hypothesize that the Hippo signaling pathway actively blocks mammalian MG-mediated retinal regeneration by preventing sustained MG proliferation and reprogramming to a retinal progenitor-like state. Our specific aims will precisely define the requirement of Hippo signaling in negative regulation of persistent cell cycle re-entry of MGs responding to retinal damage. We will also determine whether genetic bypass of Hippo signaling results in MG reprogramming to a progenitor-like state. Finally, we will assess the neurogenic potential of reprogrammed MGs. This project will employ a multi-disciplinary approach using genetic loss- and gain-of-function experiments, fate mapping, epigenomics, and single cell transcriptomics. By completion of the described aims, we expect to have identified the Hippo pathway as the endogenous molecular mechanism normally restraining MG proliferation and reprogramming to regenerative, progenitor cells. These data will provide an essential molecular entry point from which to further investigate novel methods to promote Müller glial cell-mediated retinal regeneration and likely have significant influence on the field of retinal regenerative medicine. We anticipate subsequent investigation into more translational, therapeutic methods to modulate Hippo pathway activity to promote retinal regeneration.