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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要多种人类病理会导致视网膜神经节细胞 (RGC) 变性和不可逆的视力丧失。基于干细胞的细胞替代疗法为视力恢复提供了有前景的新方法。然而，在这些疗法进行临床试验之前，还有许多问题需要克服：将胚胎干细胞 (ESC) 分化为视网膜命运会产生少量的 RGC，而之前的研究报告表明，只有一小部分 RGC 可以与宿主视网膜整合移植。本提案的主要目标是：（1）改进现有协议以有效地在最佳移植年龄将 ESC 分化为 RGC 命运，以及 (2) 识别可能的障碍到细胞植入。 1) 为了产生高产量的 ESC 衍生的 RGC：我们的实验室和其他实验室已经实施了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通过系统地测试阵列信号线索和 miRNA 调节因子来增加 RGC 的产量已知参与正常发育过程中的 RGC 分化。通过使用多种方法包括 RNA 测序和电生理学，我们将表征 ESC 衍生的 RGC。我们将标记，纯化并移植一系列不同年龄的 RGC，以确定最佳发育阶段成为移植的供体细胞。 2) 分析移植后 ESC 衍生的 RGC 的整合潜力：我们将使用状态- 最先进的成像技术可追踪活体啮齿动物移植后个体 ESC 衍生的 RGC 准确测量生存、迁移和与宿主的融合。我们将采用小鼠模型 RGC 变性以测定环境在 RGC 植入潜力中的作用。