Role of growth and differentiation factors in retinal ganglion cell development

生长和分化因子在视网膜神经节细胞发育中的作用

• 批准号: 9794004
• 负责人: Kun-Che Chang
• 金额: $ 6.37万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-24 至 2021-09-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9794004
• 关键词: Address; Adult; Affect; Alleles; Biochemical; Biological Assay; Blindness; Bone Morphogenetic Proteins; Brain; Cell Culture Techniques; Cell Death; Cell Differentiation process; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Data; Degenerative Disorder; Development; Differentiation and Growth; Disease; Down-Regulation; Embryo; Equilibrium; Eye; Flow Cytometry; GDF11 gene; Generations; Genes; Glaucoma; Goals; Hippocampus (Brain); Human; Immunofluorescence Immunologic; Injury; Knock-out; LoxP-flanked allele; Mammals; Mediating; Mentors; Microscopy; Mus; Nerve Degeneration; Nervous system structure; Neuraxis; Neurites; Neurodegenerative Disorders; Neurons; Optic Nerve; Patients; Pattern; Phosphorylation; Physiologic Intraocular Pressure; Play; Protocols documentation; Regulation; Reporting; Retina; Retinal; Retinal Ganglion Cells; Role; SOX11 gene; SOX4 gene; Signal Pathway; Signal Transduction; Stem cell transplant; Stem cells; Symptoms; Testing; Therapeutic; Time; Transforming Growth Factor beta; Transplantation; Vision; Western Blotting; Work; axon growth; base; cell fate specification; embryonic stem cell; human embryonic stem cell; human stem cells; improved; in vivo; induced pluripotent stem cell; inhibitor/antagonist; interest; member; myelination; neurite growth; neurodevelopment; neurogenesis; neuronal growth; novel; novel therapeutic intervention; optic nerve disorder; overexpression; receptor; retinal progenitor cell; retinogenesis; stem cell differentiation; transcription factor; treatment strategy; visual information

Role of Growth and Differentiation Factors in Retinal Ganglion Cell Development Glaucoma is among the leading causes of blindness, affecting roughly 60 million people worldwide.! Retinal ganglion cell and optic nerve degeneration are the major symptoms that result in permanent loss of vision in the patients. Currently, lowering intraocular pressure (IOP) is the only way to treat glaucoma; however, IOP reduction is not always sufficient to stop the underlying progression of RGC death. Understanding the intrinsic and extrinsic factors involved in RGC development and their application in cell- based therapies are critical therapeutic objectives for reversing vision loss from glaucoma and other optic neuropathies. Growth and differentiation factor 11 (GDF-11) was reported to negatively regulate RGC differentiation. Interestingly, we have found that another GDF member, GDF-15, was shown to positively regulate hippocampal neurogenesis. In retinal progenitor cell (RPC) culture, GDF-11 and GDF-15 regulate opposing RGC fate and mediate different Smad signaling. Based on our preliminary data, I hypothesize that GDF-15 promotes RGC differentiation and neurite patterning during development, and that Smad-2 signaling is important for RGC differentiation during retinal development. To address this hypothesis, I will determine the levels of RGC marker expression in PRC culture during embryonic stages in the presence of GDF-11 or GDF- 15 by qPRC and Western blot. To investigate the effects of GDFs in retinal development, I will use a floxed GDF-11 or Smad-2 allele to conditionally knock-out target genes in Chx10 expressing retinal cells at different developmental time points. In addition, I will apply GDF-15 in a novel directly induced RGC (iRGC) protocol, which will engage the differentiation of RGCs from embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs). The overall goal of this study is to better understand how GDFs influences RGC differentiation, the mechanism behind the regulation and its application to stem cell replacement, highlighting it as a potential therapeutic strategy.

生长分化因子在视网膜神经节细胞发育中的作用 青光眼是导致失明的主要原因之一，影响全球约6000万人。 视网膜神经节细胞和视神经变性是导致永久性视网膜功能丧失的主要症状。 患者的视力。目前，降低眼内压（IOP）是治疗青光眼的唯一方法; 然而，降低IOP并不总是足以阻止RGC死亡的潜在进展。 了解参与RGC发育的内在和外在因素及其在细胞中的应用- 基础疗法是逆转青光眼和其他视神经疾病引起的视力丧失的关键治疗目标。 神经病生长和分化因子11（GDF-11）被报道负调节RGC 分化有趣的是，我们发现GDF的另一个成员GDF-15， 调节海马神经发生。在视网膜祖细胞（RPC）培养物中，GDF-11和GDF-15调节视网膜祖细胞的生长。 与RGC命运相反，并介导不同的Smad信号传导。根据我们的初步数据，我假设 GDF-15在发育过程中促进RGC分化和神经突形成，而Smad-2信号传导是促进RGC分化和神经突形成的重要因素。 在视网膜发育过程中对RGC分化起重要作用。为了解决这个假设，我将确定 在GDF-11或GDF-11存在下，在胚胎阶段期间PRC培养物中RGC标志物表达水平 15通过qPRC和Western印迹。为了研究GDFs在视网膜发育中的作用，我将使用一个floxed GDF-11或Smad-2等位基因在不同浓度下条件性敲除表达Chx 10的视网膜细胞中的靶基因 发展时间点。此外，我将在一种新的直接诱导RGC（iRGC）方案中应用GDF-15， 其将参与来自胚胎干细胞（ESC）或诱导多能干细胞（MSC）的RGCs的分化。 细胞（iPSC）。本研究的总体目标是更好地了解GDF如何影响RGC分化， 调控背后的机制及其在干细胞替代中的应用，强调它是一种潜在的 治疗策略