Derivation and disease modeling of human stem cell-derived retinal ganglion cells

人类干细胞来源的视网膜神经节细胞的衍生和疾病模型

• 批准号: 9187803
• 负责人: Jason Stephen Meyer
• 金额: $ 35.45万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9187803
• 关键词: Affect; Allogenic; Alpha Cell; Apoptosis; Apoptotic; Astrocytes; Attention; Autologous; Biological Assay; Biological Models; Blindness; Cell Cycle; Cell Differentiation process; Cell Survival; Cells; Characteristics; Coculture Techniques; Cues; Degenerative Disorder; Derivation procedure; Development; Disease; Disease model; Electrophysiology (science); Event; Exhibits; Exposure to; Genes; Genetic; Glaucoma; Growth Factor; Human; Hydrogen Peroxide; In Vitro; Injury; Lead; Measurement; Methods; Mutation; Nature; Nerve Degeneration; Ocular Hypertension; Patients; Pharmacologic Substance; Phenotype; Photoreceptors; Physiologic Intraocular Pressure; Population; Predisposition; Primary Open Angle Glaucoma; Proteins; Protocols documentation; Reactive Oxygen Species; Research; Resistance; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Retinal Ganglion Cells; Retinal Pigments; Role; Sampling; Somatic Cell; Source; Stem cells; Stress; Study models; Symptoms; System; Testing; base; cell injury; cell type; cytokine; design; disease phenotype; environmental stressor; experimental study; human stem cells; immunocytochemistry; in vitro Model; induced pluripotent stem cell; novel; novel strategies; novel therapeutics; optic nerve disorder; public health relevance; response; retinal progenitor cell; small molecule inhibitor; stressor

DESCRIPTION (provided by applicant): Human induced pluripotent stem cells (hiPSCs) provide a unique source of stem cells that have the potential to differentiate into any cell type o the body, including retinal ganglion cells. Furthermore, when derived from a patient's own somatic cells, they have the potential to faithfully serve as a model system of glaucoma. Previously, we have shown the ability to derive various retinal cell types including retinal ganglion cells from hiPSCs. In the current proposal, hiPSCs will be derived from patients with normal-tension primary open angle glaucoma, which would constitute a source of cells with a high predisposition to retinal ganglion cell damage. Conversely, wild type hiPSCs will be utilized as a point of comparison which would have a greater resistance to glaucomatous injury. The comparison of retinal ganglion cells derived from these two sources will allow for the identification of factors contributing to glaucoma-related symptoms. To test this hypothesis, retinal ganglion cells derived from both sources will be examined for the accumulation of reactive oxygen species characteristic of a glaucomatous condition, both under control conditions as well as after exposure to environmental stressors such as hydrogen peroxide. Viability of these cells will also be tested through the use of apoptotic assays to identify differences between the two populations. It is expected that such stressors will result in increased apoptotic activity in those cells derived from glaucoma patients compared with those derived from ocular hypertension sources. Additionally, the role of astrocytes in the progression of a glaucomatous phenotype will be tested. Astrocytes will be derived from patient-specific hiPSCs and co-cultured with retinal ganglion cells. Analysis will be performed to determine the extent of astrocyte contribution to the glaucoma disease phenotype. The successful completion of the aims of this proposal will establish patient-specific hiPSC- derived retinal ganglion cells s a valuable model system for studies of glaucoma, and will also serve as a significant basis upon which to design new pharmaceuticals for the treatment of this disease.

描述(申请人提供)：人类诱导多能干细胞(HiPSCs)提供了一种独特的干细胞来源，有可能分化为身体内任何类型的细胞，包括视网膜神经节细胞。此外，当它们来自患者自己的体细胞时，它们有可能忠实地充当青光眼的模型系统。在此之前，我们已经展示了从HiPSC分化出各种类型的视网膜细胞的能力，包括视网膜神经节细胞。在目前的提议中，HiPSCs将来自正常眼压性原发性开角型青光眼患者，这将构成视网膜神经节细胞损伤的高易感性细胞来源。相反，野生型HiPSCs将被用作对青光眼损伤具有更强抵抗力的比较点。对来自这两个来源的视网膜神经节细胞进行比较，将有助于识别导致青光眼相关症状的因素。为了验证这一假设，我们将检查来自这两个来源的视网膜神经节细胞在控制条件下以及暴露于过氧化氢等环境应激源后，青光眼条件下特有的活性氧物种的积累情况。这些细胞的活性也将通过使用凋亡分析来测试，以确定两个群体之间的差异。预计这种应激源将导致青光眼患者来源的细胞与高眼压来源的细胞相比，细胞凋亡活性增加。此外，还将测试星形胶质细胞在青光眼表型进展中的作用。星形胶质细胞将从患者特有的HiPSCs中分离出来，并与视网膜神经节细胞共同培养。将进行分析以确定星形胶质细胞对青光眼疾病表型的贡献程度。这项提案目标的成功完成将使患者特异性的HIPSC来源的视网膜神经节细胞S成为青光眼研究的一个有价值的模型系统，并将作为设计治疗这种疾病的新药的重要基础。