Regulation of interneuron formation in the developing retina

视网膜发育中中间神经元形成的调节

• 批准号: 10677004
• 负责人: DAVID L TURNER
• 金额: $ 42.76万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10677004
• 关键词: Amacrine Cells; Binding Proteins; Binding Sites; Biological Assay; Biology; Birth; Blindness; CRISPR/Cas technology; Cell Differentiation process; Cells; Central Nervous System; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA Binding; Development; Developmental Process; Ectopic Expression; Embryonic Development; Eye; Family; Gene Expression Profile; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Genomics; Goals; Health; Health Care Costs; Human; Individual; Inherited; Injury; Interneurons; Intervention; Knowledge; Link; Mammals; Mediating; Messenger RNA; Methodology; MicroRNAs; Molecular; Mus; Natural regeneration; Neural Crest; Neurons; Pathway interactions; Patients; Proteins; Quality of life; RNA Interference; Regulation; Reporter; Reporting; Repression; Retina; Retinal Diseases; Role; Site; Structure; Testing; Tissues; Transcription Coactivator; Transcription Repressor; Variant; Visual Perception; Visual impairment; Work; cell determination; cell type; cholinergic; data integration; economic cost; gene repression; gene therapy; genetic variant; insight; mRNA Expression; novel strategies; overexpression; postmitotic; repair strategy; repaired; retinal regeneration; retinogenesis; stem cell biology; stem cells; transcription factor; transcriptome sequencing

Abstract/Project Summary Retinal disease or injury leading to impaired vision or blindness are human health problems that reduce quality of life, generating significant human and economic costs. Advances in gene therapy and stem cell biology have made retinal repair a feasible goal. Nonetheless, rational repair strategies are constrained by current knowledge of retinal biology and development. The complex cellular composition of the retina, like other parts of the mammalian central nervous system, is an essential component of the retina’s functional capabilities, yet remains incompletely understood. The mammalian retina includes more than 100 distinct types of neurons. The generation of this cellular diversity during retinal development depends in part on sequential cascades of transcriptional regulators as well as other factors. We found the miR-216b microRNA can influence retinal development including the formation of amacrine and bipolar interneurons. We identified a target gene for this microRNA, a transcriptional repressor in the forkhead family, Foxn3, that when inhibited using RNAi or CRISPR in the developing retina increases amacrine cell formation, and when overexpressed reduces amacrine cell formation. The target genes of Foxn3 during retinal development are not known. Here we propose to identify mRNAs regulated by either loss or gain of Foxn3 function in the developing retina. We will analyze changes in mRNA expression to identify the molecular pathways by which Foxn3 modulates retinal cell fate decisions. In addition, we will identify genomic sites at which the Foxn3 protein binds in the developing retina to find candidate target genes. We also propose to analyze retinal development in the absence of miR- 216b or the related miR-216a microRNA, to determine if these miRNAs are required for amacrine cell differentiation or other functions, and to assess the function of two miR-216a/b genetic variants that may be linked to retinal disease. These studies will provide new insights into the regulation of development and cell determination in the mammalian retina. They are expected to provide information that may be relevant to retinal disease and that may contribute to new strategies to repair retinal tissue.

摘要/项目摘要 导致视力受损或失明的视网膜疾病或损伤是人类健康问题， 生活质量，产生巨大的人力和经济成本。基因治疗与干细胞研究进展 生物学使视网膜修复成为一个可行的目标。尽管如此，合理的修复策略受到以下因素的限制： 视网膜生物学和发育的现有知识。视网膜的复杂细胞组成， 哺乳动物中枢神经系统的其他部分，是视网膜功能的重要组成部分， 能力，但仍不完全了解。哺乳动物的视网膜包括100多种不同的类型 的神经元。在视网膜发育过程中，这种细胞多样性的产生部分取决于连续的 转录调节因子以及其他因子的级联反应。我们发现miR-216 b microRNA可以 影响视网膜发育，包括无长突和双极中间神经元的形成。我们确定了一个 这种微小RNA的靶基因，叉头家族中的转录抑制因子Foxn 3， 在发育中的视网膜中使用RNAi或CRISPR增加了无长突细胞的形成，当过度表达时， 减少无长突细胞的形成。Foxn 3在视网膜发育过程中的靶基因尚不清楚。这里 我们建议鉴定发育中视网膜中Foxn 3功能丧失或获得所调控的mRNA。我们 将分析mRNA表达的变化，以确定Foxn 3调节视网膜病变的分子途径。 细胞命运的决定此外，我们将确定Foxn 3蛋白在发育中结合的基因组位点。 视网膜寻找候选靶基因。我们还建议分析视网膜发育的情况下，miR- 216 b或相关的miR-216 a microRNA，以确定这些miRNAs是否是无长突细胞所必需的。 分化或其他功能，并评估两种miR-216 a/B遗传变异体的功能， 与视网膜疾病有关。这些研究将为发育和细胞生长的调控提供新的见解。 在哺乳动物视网膜中的决定。他们应提供可能与下列事项有关的资料： 这可能有助于新的策略来修复视网膜组织。