Regulation of Human Embryonic Stem cell Neuro-retinal Differentiation

人胚胎干细胞神经视网膜分化的调控

• 批准号: 8460658
• 负责人: THOMAS A REH
• 金额: $ 27.81万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-12-17 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8460658
• 关键词: Accounting; Affect; Age; American; Area; Biomedical Research; Caenorhabditis elegans; Cell Line; Cell Therapy; Cells; Cultured Cells; Data; Defect; Degenerative Disorder; Development; Disease; Disease model; Engineering; Event; Failure; Gene Expression Profiling; Genes; Health; Human; In Vitro; Individual; Inherited; Macular degeneration; Messenger RNA; MicroRNAs; Modeling; Muller' s cell; Mus; Neuroglia; Organism; Pathway interactions; Patients; Phase; Population; Production; Protocols documentation; Regulation; Research; Retina; Retinal; Retinal Cone; Retinal Diseases; Staging; Stem Cell Research; Stem cells; Testing; Time; Vision; age related; cell type; embryonic stem cell; fetal; fetus cell; ganglion cell; histogenesis; horizontal cell; human DICER1 protein; human embryonic stem cell; in vivo; induced pluripotent stem cell; interest; mouse model; overexpression; postnatal; progenitor; research study; retinal progenitor cell; retinal rods; self-renewal; sensor

One of the most important potential uses of embryonic stem cells and induced pluripotent stem cell research is in the development of disease models. Although many diseases can be modeled in mice, others are difficult or impossible to adequately model in non-human organisms for a variety of reasons. In the retina, for example, there are mouse models for many inherited degenerations, but these have been more difficult to generate for diseases like macular degeneration and many forms of Usher's disease. Therefore, disease models from human patient iPS cells would be of great utility in retinal research. In the past six years, our group, and others, have developed protocols for efficient production of retinal cells from human ESCs and iPSCs. The early stages of retinal development are extremely well recapitulated by our protocol of directed differentiation of hESC cells; however, the cells fail to attain the level of expression of markers that we observe in the late stages of human fetal or postnatal development, even with prolonged culture periods. Therefore, a significant challenge for creation of disease models from iPSCs will be to better understand and promote the progression of the cultured cells to mature stages of retina. The mechanisms that control the developmental timing of retinal progenitor cells are not clear; however, recent evidence from our lab shows a key role for miRNAs. Specifically, we found that the loss of Dicer in retinal progenitor cells leads to their failure to progress from the "early" state to the "late" state. We therefore hypothesize (1) that miRNAs regulate developmental timing in retinal progenitor cells and (2) that misregulation of the heterochronic pathway in ESC-derived retinal cells accounts for their failure to progress in vitro at the same rate as in vivo. We propose to test these hypotheses with the following specific aims. Aim 1: Determine whether the expression of specific miRNAs (and the genes they regulate) correlates with the progression of retinal progenitors from the early to late stage. Aim 2.Determine whether the developmental progression of mouse ESC/iPSC-derived retinal progenitors is controlled by stage-specific progenitor miRNAs. Aim 3. Determine whether the developmental progression of human ESC derived retinal progenitors can be accelerated by stage-specific miRNAs. The results of these studies will enable us to better control the development of hESC-derived retinal cells and produce more appropriate models of retinal disease.

胚胎干细胞和诱导多能干细胞研究的最重要潜在用途之一是疾病模型的发展。尽管许多疾病可以在小鼠中进行建模，但由于各种原因，其他疾病很难或不可能在非人类生物中进行充分建模。例如，在视网膜中，有许多鼠标用于许多遗传的变性，但这些模型更难生成 诸如黄斑变性和多种形式的usher病等疾病。因此，来自人类患者IPS细胞的疾病模型在视网膜研究中将非常有用。在过去的六年中，我们的小组和其他人开发了有效生产人类ESC和IPSC的视网膜细胞的方案。视网膜发育的早期阶段被我们的定向分化方案很好地概括了。但是，即使培养期延长，这些细胞无法达到我们在人类胎儿或产后发育的后期观察到的标志物表达水平。因此，从IPSC创建疾病模型的重大挑战将是更好地理解和促进培养细胞到视网膜成熟阶段的发展。控制视网膜祖细胞的发育时机的机制尚不清楚。但是，我们实验室的最新证据显示了miRNA的关键作用。具体而言，我们发现视网膜祖细胞中的DICER损失导致它们未能从“早期”状态发展到“晚期”状态。因此，我们假设（1）miRNA调节发育时机 视网膜祖细胞和（2）ESC衍生的视网膜细胞中异缘途径的正调是由于其在体外无法在体外进展的速度与体内相同的速度。我们建议以以下特定目的检验这些假设。 目标1：确定特定miRNA（及其调节基因）的表达是否与视网膜祖细胞从早期到晚期的进展相关。 AIM 2.确定是否由小鼠ESC/IPSC衍生的视网膜祖细胞的发育进展是否受阶段特异性的祖细胞miRNA控制。目标3。确定人ESC衍生的视网膜祖细胞的发育进展是否可以通过特定于阶段的miRNA加速。这些研究的结果将使我们能够更好地控制hESC衍生的视网膜细胞的发展，并产生更合适的视网膜疾病模型。