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.

项目摘要/摘要缺乏合适的系统可以阻碍治疗视网膜退行性疾病评估新的治疗策略如何影响人类视网膜的功能。人类干细胞衍生的3- 尺寸视网膜器官技术最近已开发。值得注意的是，人类视网膜器官模仿天然组织的组织学组织，细胞组成，并能够对光反应。这些器官是研究治疗盲疾病的新型疗法的理想模型系统。但是，要充分意识退化性疾病，我们需要新技术，这些技术可以在各种各样的种类下快速测量视网膜功能条件。当前技术，例如贴片钳电生理学，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作用。但是，这些技术在评估视网膜的健康，可重复性和功能反应性方面努力且无效整个功能是将器官系统应用于药物筛查和验证的关键。缺乏一种设备和技术，可以快速，无创筛选视网膜类器官的功能状态构成了主要的未满足需求。在本提案的目标1中，我们将开发电子图（ERG）记录腔室和记录方案，用于实时评估光诱发的视网膜器官生理学测量光感受器和双极细胞功能。我们的发现将用于建立关键指标与正常的器官诱发的响应性相关。此外，我们将评估这种方法的力量检测光感受器功能的变化，以提供其适用于疾病评估的证据模型和治疗性筛查。为了充分利用这项用于下游应用的技术，至关重要健壮的器官模型。视网膜器官产生的当前方案的可变性和低收益率和低收率功能性成熟器官光感受器所需的延长时间阻碍了它们在药物中的应用发展和疾病评估。在该提案的目标2中，我们将通过发展来解决这一关键差距并评估改进人类视网膜器官生成的改进方案，以提高产量和加速光感受器的分化。然后，我们将使用我们的ERG评估这些改进的器官中的视网膜功能平台。通过这些技术的结合，我们将创建第一个用于快速，非 - 在人类视网膜器官中的侵入性功能筛查，可以应用于正常的评估患病和药物治疗的疾病。我们的系统有可能大大加速逆转视力丧失的新疗法。