权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Novel tools for screening retinal function using improved human retinal organoid models

使用改进的人类视网膜类器官模型筛查视网膜功能的新工具

基本信息

批准号：
10462668
负责人：
Jordan Michael Renna
金额：
$ 11.3万
依托单位：
UNIVERSITY OF AKRON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10462668
关键词：
3-Dimensional 9-cis-retinal Address Affect Biological Assay Biological Models Blindness Calcium Cell physiology Cone Development Devices Disease Disease model Drug Screening Electrophysiology (science)Electroretinography Evaluation Gene Expression Generations Health Histologic Human Human Development Image Individual Light Measures Modeling Organoids Pharmaceutical Preparations Pharmacological Treatment Pharmacotherapy Photoreceptors Physiological Physiology Protocols documentation Reproducibility Research Personnel Retina Retinal Cone Retinal Degeneration Retinal Diseases Screening procedure Signal Transduction Source Speed Supplementation System Techniques Technology Testing Therapeutic Therapeutic Uses Time Tissues Tretinoin Validation Work cost drug development experimental study functional status human stem cells improved interdisciplinary approach interest multi-electrode arrays neurophysiology new technology non-invasive system notch protein novel novel therapeutics patch clamp screening sight restoration small molecule stem cells therapy development tool transcriptomics treatment strategy

项目摘要

Project Summary/Abstract Treating retinal degenerative diseases has been hampered by the lack of suitable systems that can evaluate how new treatment strategies affect the function of the human retina. Human stem cell-derived 3- dimensional retinal organoid technologies have been recently developed. Remarkably, human retinal organoids mimic the native tissue's histological organization, cellular composition, and are able to respond to light. These organoids are an ideal model system for investigating novel therapies to treat blinding diseases. However, to fully realize the unprecedented potential of human retinal organoids for the development of treatments for retinal degenerative diseases, we need new technologies that can rapidly measure retinal function under a wide variety of conditions. Current techniques such as patch-clamp electrophysiology, calcium imaging, and multi-electrode array recordings, measure how individual cells function within the circuitry of the retina. However, these techniques are laborious and ineffective at assessing the health, reproducibility, and functional responsivity of the retina as a whole - features that are key to the application of organoid systems to drug screening and validation. The lack of a device and techniques that allow for rapid, non-invasive screening of the functional status of retinal organoids constitutes a major unmet need. In Aim 1 of this proposal, we will develop an electroretinogram (ERG) recording chamber and a recording protocol for real-time assessment of light-evoked retinal organoid physiology by measuring photoreceptor and bipolar cell function. Our findings will be used to establish critical metrics associated with normal organoid light-evoked responsivity. Further, we will evaluate the power of this approach to detect changes in photoreceptor function, to provide evidence of its applicability to the assessment of disease models and therapeutic screening. To take full advantage of this technology for downstream applications it is critical that it be combined with robust organoid models. The variability and low yield of current protocols for retinal organoid generation and the extended time required for functional maturation of organoid photoreceptors hinder their application in drug development and disease evaluation. In Aim 2 of this proposal, we will address this critical gap by developing and evaluating improved protocols for human retinal organoid generation that increase yield and accelerate photoreceptor differentiation. We will then evaluate retinal function in these improved organoids using our ERG platform. Through the combination of these technologies, we will have created the first system for rapid, non- invasive functional screening in human retinal organoids that can be applied to the evaluation of normal, diseased, and drug-treated conditions. Our system has the potential to greatly accelerate the development of novel therapies to reverse vision loss.

项目总结/摘要视网膜变性疾病的治疗一直受到缺乏合适系统的阻碍，评估新的治疗策略如何影响人类视网膜的功能。人干细胞来源的3- 最近开发了三维视网膜类器官技术。值得注意的是，人类视网膜类器官模仿天然组织的组织学组织、细胞组成，并且能够对光做出反应。这些类器官是研究治疗致盲性疾病的新疗法的理想模型系统。但要充分认识到人类视网膜类器官在开发视网膜病变治疗方面的前所未有的潜力，对于退行性疾病，我们需要新的技术，可以在各种各样的条件下快速测量视网膜功能。的条件。目前的技术，如膜片钳电生理学，钙成像，和多电极阵列记录，测量单个细胞在视网膜电路中的功能。然而，这些技术在评估视网膜的健康、再现性和功能响应性方面是费力和无效的，一个整体-特征是类器官系统应用于药物筛选和验证的关键。缺乏允许快速、非侵入性筛查视网膜类器官功能状态的设备和技术这是一个未满足的主要需求。在本提案的目标1中，我们将开发一种视网膜电图（ERG）记录室和记录协议的实时评估光诱发的视网膜类器官生理测量光感受器和双极细胞的功能。我们的研究结果将用于建立关键指标与正常的类器官光诱发反应有关。此外，我们将评估这种方法的功效检测光感受器功能的变化，提供其适用于疾病评估的证据模型和治疗筛选。为了充分利用这项技术在下游应用中的优势，将其与以下技术相结合至关重要：鲁棒的类器官模型。目前用于视网膜类器官产生的方案的可变性和低产量以及类器官光感受器功能成熟所需的时间延长阻碍了它们在药物中的应用发展和疾病评估。在本提案的目标2中，我们将通过开发以及评估用于人类视网膜类器官产生的改进方案，感光细胞分化然后，我们将使用我们的ERG评估这些改善的类器官中的视网膜功能平台通过这些技术的结合，我们将创造出第一个快速、非在人类视网膜类器官中进行侵入性功能筛选，疾病和药物治疗的情况。我们的系统有潜力大大加快发展，逆转视力丧失的新疗法。