Induced Pluripotent Stem Cell Approach to Glaucomatous Optic Neuropathy

诱导多能干细胞治疗青光眼视神经病变

基本信息

批准号：
8437370
负责人：
Iqbal Ahmad
金额：
$ 37.13万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-12-01 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8437370
关键词：
Acids Address Adult Affect African American American Autologous Axon Biological Models Blindness Cell Differentiation process Cell Lineage Cell Therapy Cells Clinic Clinical Coculture Techniques Conditioned Culture Media Cornea Cues Data Development Embryo Ensure Exogenous Factors Experimental Models Eye Fibroblast Growth Factor Generations Genes Genetic Glaucoma Goals Home environment In Vitro Insertional Mutagenesis Lateral Geniculate Body Lead Link Mediating Methods Midbrain structure Molecular Natural regeneration Neurons Nucleic Acids Pathway interactions Perception Physiologic Intraocular Pressure Pluripotent Stem Cells Preclinical Drug Evaluation Property Rattus Reagent Recruitment Activity Relative (related person)Research Retina Retinal Retinal Degeneration Retinal Ganglion Cells Risk Signal Pathway Signal Transduction Somatic Cell Source Specificity Stem cells Testing Transplantation Vision Visual Visual Pathways Work area striata axonal guidance base drug testing effective therapy gene therapy in vivo induced pluripotent stem cell innovation limbal nerve stem cell notch protein novel strategies optic nerve disorder pluripotency prevent progenitor public health relevance relating to nervous system response retinal progenitor cell small molecule stem cell therapy success superior colliculus Corpora quadrigemina

项目摘要

DESCRIPTION (provided by applicant): Glaucoma is the most prevalent optic neuropathy where a progressive degeneration of retinal ganglion cells (RGCs) leads to vision loss. Our long-term goal is to help prevent the degeneration of glaucomatous RGCs by characterizing induced-pluripotent stem cells (iPSCs) as a renewable source of retinal progenitors for autologous ex vivo cell therapy. The objective of this application is to optimize the use of limbal iPSCs to generate RGCs that are functional, safe, and practical for clinical use. The central hypothesis of the proposed study is that the molecular mechanism underlying RGC differentiation is active in iPSC-derived retinal progenitors and recruited in response to specific extrinsic cues to generate RGCs with target specificity. Our reasoning is based on the following observations:(1) retinal progenitors can be derived from limbal iPSCs, generated through safe non-nucleic acid method (2) iPSC-derived retinal progenitors respond to cues conducive for RGC differentiation, and (3) iPSC-derived RGCs demonstrate target specificity. The rationale for the proposed research is that once conditions are identified, we can efficiently generate RGC precursors to treat RGC degeneration through transplantation, and develop a robust model system for testing drugs and genetic approaches for optic neuropathy. Based on our preliminary data the following specific aims are proposed to test the hypothesis: Specific Aim 1: To determine the conditions for generating retinal progenitors from iPSCs, Specific Aim 2: To determine conditions for the generation of RGCs from iPSC-derived retinal progenitors, and Specific Aim 3: To determine the target specificity and in vivo differentiation of iPSC-derived RGCs. The retinal potential will be examined in limbal iPSCs generated by non-nucleic acid means, pioneered in our lab. This approach of reprogramming by recruiting endogenous pluripotency genes instead of introducing exogenous genes, which can lead to insertional mutagenesis, addresses a significant barrier to iPSC-based therapy. Controls will include limbal iPSC derived by a conventional nucleic acid method to compare the effects of two different approaches of reprogramming on the acquisition of retinal and RGC potential. The induction of iPSCs along a neural lineage, their subsequent specification into retinal progenitors, and their final differentiation into RGCs will be accomplished non-cell autonomously by perturbing specific signaling pathways to recapitulate developmental mechanism. Therefore, our research proposed is innovative because it presents an entirely different and a safe approach for reprogramming somatic cells to a pluripotent state and generating RGCs without using nucleic acids or forced expression of exogenous factors. The emerging information will be significant because it will not only address each of the barriers that currently make the ex-vivo stem cell therapy approach impractical but also lead to the development of a robust model system for testing normal mechanisms of RGC development and for screening drugs and genes for additional new approaches for addressing glaucomatous retinal degeneration.

描述（由申请人提供）：青光眼是最普遍的视神经神经病，其中视网膜神经节细胞（RGC）的进行性变性会导致视力丧失。我们的长期目标是通过将诱导的血管干细胞（IPSC）表征为自体外细胞疗法的视网膜祖细胞的可再生能源来帮助防止青光眼RGC的变性。此应用程序的目的是优化缘缘的使用 IPSC生成功能性，安全且可用于临床使用的RGC。拟议的研究的中心假设是，RGC分化的分子机制在IPSC衍生的视网膜祖细胞中活跃，并响应特定的外部线索，以产生具有目标特异性的RGC。我们的推理基于以下观察结果：（1）视网膜祖细胞可以源自通过安全的非核酸方法生成的层层IPSC（2）IPSC衍生的视网膜祖细胞对有利于RGC分化的提示的响应，并且我们（3）IPSC衍生的RGC均表明靶向靶标。拟议的研究的理由是，一旦确定了条件，我们就可以通过移植有效地生成RGC前体来治疗RGC变性，并开发出可靠的模型系统，用于测试药物和遗传方法的视神经病变。 Based on our preliminary data the following specific aims are proposed to test the hypothesis: Specific Aim 1: To determine the conditions for generating retinal progenitors from iPSCs, Specific Aim 2: To determine conditions for the generation of RGCs from iPSC-derived retinal progenitors, and Specific Aim 3: To determine the target specificity and in vivo differentiation of iPSC-derived RGCs.视网膜电位将以非核酸均值产生的缘含IPSC进行检查，并在我们的实验室中率先研究。通过募集内源性多能基因而不是引入外源基因来重新编程的方法，这可能导致插入诱变，它涉及基于IPSC治疗的重大障碍。对照将包括通过常规核酸方法得出的缘含IPSC，以比较两种不同的重编程方法对获得视网膜和RGC电位的采集的影响。 IPSC沿着神经谱系的诱导，随后将其指定为视网膜祖细胞，并将其最终分化为RGC，将通过扰动特定的信号通路来概括发育机制来实现非单元的自主。因此，我们提出的研究具有创新性，因为它提出了一种完全不同且安全的方法，可将体细胞重编程到多能状态并生成RGC，而无需使用核酸或强迫表达外源性因子。新兴信息将是重要的，因为它不仅将解决当前使前病毒细胞疗法方法不切实际的每个障碍，而且还导致开发了一种可靠的模型系统，用于测试RGC开发的正常机制，以及用于筛查药物和基因的其他新方法，用于解决glaucomatos glaucomatos retaucomatios retaucomatios retaucomatios。