Embryonic Stem Cell Approach to Retinal Ganglion Cell Replacement: an In Vivo Study

胚胎干细胞替代视网膜神经节细胞的方法：体内研究

• 批准号: 9319272
• 负责人: Anna La Torre
• 金额: $ 39.25万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9319272
• 关键词: Affect; Age; Alpha Cell; Biological Assay; Blindness; Cell Death; Cell Differentiation process; Cell Survival; Cell Transplantation; Cell Transplants; Cells; Clinical Trials; Cues; Data; Development; Disease; Electrophysiology (science); Embryonic Stem Cell Transplantation; Engraftment; Environment; Generations; Glaucoma; Goals; Growth Factor; Human; Human Pathology; Image; Imaging technology; Individual; Inner Limiting Membrane; Intervention; Label; Laboratories; Lasers; Lead; Maintenance; Measures; Methods; MicroRNAs; Molecular; Morphology; Mus; Neuroglia; Neurons; Ophthalmoscopy; Optical Coherence Tomography; Pharmaceutical Preparations; Photoreceptors; Population; Probability; Production; Prosthesis; Protocols documentation; Replacement Therapy; Reporting; Resolution; Resources; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Retinal Ganglion Cells; Rodent; Role; Scanning; Signal Transduction; Signaling Molecule; Specific qualifier value; Stem cells; Synapses; Technology; Testing; Therapeutic; Transplantation; Vision; adaptive optics scanning laser ophthalmoscopy; axon growth; base; effective therapy; embryonic stem cell; experimental study; improved; in vivo; in vivo imaging; migration; mouse model; multimodality; novel; novel strategies; optic nerve disorder; progenitor; repaired; restoration; retinal neuron; retinal progenitor cell; stem cell differentiation; success; transcriptome sequencing

Project Summary Several human pathologies lead to retinal ganglion cell (RGC) degeneration and irreversible vision loss. Stem cell-based cell replacement therapy offers promising novel approaches for vision restoration. However, there are many issues to overcome before these therapies can be clinically trialled: The current methods to differentiate embryonic stem cells (ESCs) into retinal fates yield modest numbers of RGCs, and previous reports have suggested that only a small fraction of RGCs can integrate with a host retina upon transplantation. The main goals of the present proposal are: (1) to improve the current protocols to efficiently differentiate ESCs into RGC fates at the optimal age for transplantation, and (2) to identify possible barriers to cell engraftment. 1) To generate high yields of ESC-derived RGCs: Our lab and others have implemented methods to differentiate embryonic stem cells into bona-fide retinal progenitors. We will develop the current protocols to increase the production of RGCs by systematically testing an array signaling cues and miRNA regulators known to be involved in RGC differentiation during normal development. By using a variety of approaches including RNA-sequencing and electrophysiology, we will characterize the ESC-derived RGCs. We will label, purify and transplant RGCs from an array of different ages to pinpoint the optimal developmental stage to become donor cells for transplantation. 2) To analyze integration potential of ESC-derived RGCs following transplantation: We will use state- of-the-art imaging technologies to follow individual ESC-derived RGCs after transplantation in living rodents to accurately measure survival, migration and integration with the host. We will employ murine models of RGC degeneration to assay the role of the environment in RGC engraftment potential.

项目摘要 多种人类疾病导致视网膜神经节细胞(RGC)变性和不可逆转的视力丧失。 基于干细胞的替代疗法为视力恢复提供了很有前途的新方法。然而， 在这些疗法可以临床试验之前，有许多问题需要克服：目前的方法 将胚胎干细胞(ESCs)分化为视网膜命运可以产生适度数量的RGC，而以前 有报道称，只有一小部分RGC可以与宿主视网膜结合。 移植。本提案的主要目标是：(1)改进现有的协议，使其更有效地 在移植的最佳年龄将胚胎干细胞分化为RGC命运，以及(2)确定可能的障碍 到细胞移植。 1)为了产生ESC来源的RGC的高产量：我们的实验室和其他人已经实现了以下方法 将胚胎干细胞分化为真正的视网膜祖细胞。我们将开发当前的协议以 通过系统地测试信号信号和miRNA调节器阵列来增加RGC的产量 已知在正常发育过程中参与RGC分化。通过使用各种方法 包括RNA测序和电生理学，我们将对ESC来源的RGCs进行表征。我们会贴上标签， 从不同年龄的人中提纯和移植视网膜神经节细胞，以确定最佳发育阶段 成为移植的供体细胞。 2)分析胚胎干细胞来源的视网膜节细胞移植后的整合潜能：我们将使用状态- 最先进的成像技术在活体啮齿动物体内移植后追踪单个胚胎干细胞来源的视网膜节细胞 以准确测量生存、迁徙和与宿主的融合。我们将使用小鼠模型 RGC退变检测环境对RGC植入潜能的影响。