Diversification of cell types in the Drosophila retina - Resubmission - 1

果蝇视网膜细胞类型的多样化 - 重新提交 - 1

• 批准号: 10328555
• 负责人: Jessica E Treisman
• 金额: $ 24.66万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10328555
• 关键词: Activities of Daily Living; Adenine; Binding Sites; Cell Differentiation process; Cells; Complex; Cone; DNA; Data Set; Development; Developmental Biology; Differentiated Gene; Drosophila eye; Drosophila genus; EGFR gene; Elements; Enhancers; Epidermal Growth Factor Receptor; Eye; Eye Development; Eye diseases; Feeds; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Transcription; Genomics; Glass; Goals; Individual; Injury; Light; Mediating; Methods; Methyltransferase; Mitotic; Molecular; Neural Retina; Neuroglia; Neurons; Nucleic Acid Regulatory Sequences; Organ; Output; Pathway interactions; Pattern; Photoreceptors; Pigments; Process; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Regenerative Medicine; Regulation; Retina; Retinal Cone; Signal Pathway; Signal Transduction; Specific qualifier value; Specificity; System; Testing; To specify; Transcription Repressor; Undifferentiated; Vision; Work; Zinc Fingers; cell fate specification; cell replacement therapy; cell type; design; experimental study; extracellular; fly; genetic analysis; improved; lens; mutant; prevent; progenitor; programs; retinal regeneration; screening; stem cells; transcription factor; transcriptomics

Summary The development of a complex organ such as the eye involves the specification of multiple distinct cell types and the integration of their functions. In Drosophila, the light-detecting photoreceptors and the glial-like cone and pigment cells that secrete the lens and produce screening pigments arise from a single field of equivalent progenitors. Commitment to one of these cell fates requires extracellular signals to be integrated with a complex network of intrinsic transcription factors. Although receptor tyrosine kinase signaling has been known for many years to induce both photoreceptors and cone cells, most of its target genes remain unknown. It is also unclear which intrinsic transcription factors distinguish these cell identities. The zinc finger transcription factor Glass had been thought to specify the photoreceptor fate, but our recent work showed that it acts in each of the three cell types to promote their normal differentiation. This proposal seeks to understand how these common factors activate distinct differentiation pathways. The first aim will look for direct target genes of receptor tyrosine kinase signaling and Glass during retinal differentiation. Targeted DNA adenine methyltransferase identification (DamID) will be used to identify direct target genes of Pointed, the transcription factor that mediates Epidermal growth factor receptor (EGFR) signaling, in undifferentiated cells, photoreceptors and cone cells. A similar approach will be used to identify Glass target genes, and both datasets will be compared to transcriptomic analysis of genes that change their expression in Egfr or glass mutants. The goal of these experiments is to identify transcription factors that are induced by EGFR signaling to promote the differentiation of photoreceptors and cone cells, and elucidate how Glass feeds into their regulation. The second aim concerns how transcriptional repressors restrict the effects of Glass to drive cell type-specific gene expression. Two defined regulatory regions that are known to drive photoreceptor-specific expression in a Glass-dependent manner will be used to identify repressors that prevent Glass from activating these genes in cone and pigment cells. The importance of these repressor binding sites will be tested in the endogenous genomic context. In a complementary approach, enhancer regions that drive Glass-dependent expression specifically in cone or pigment cells will be identified and their regulation characterized. In combination, these experiments will reveal how distinct cell identities can be specified from common progenitors using a limited set of signals and transcription factors, a process that also occurs in the mammalian retina. The results will help to refine our ability to produce specific cell types from stem cells for regenerative medicine.

概括 眼睛等复杂器官的发育涉及多个不同的规范 细胞类型及其功能的整合。在果蝇中，光检测光感受器和 分泌晶状体并产生筛选色素的神经胶质样锥细胞和色素细胞来自于单个细胞 同等祖细胞的领域。对这些细胞命运之一的承诺需要细胞外信号 与内在转录因子的复杂网络整合。虽然受体酪氨酸激酶 多年来，人们都知道信号传导可以诱导光感受器和视锥细胞，这是其大部分目标 基因仍然未知。还不清楚哪些内在转录因子区分这些细胞身份。 锌指转录因子 Glass 曾被认为决定了光感受器的命运，但我们最近的研究 研究表明，它在三种细胞类型中发挥作用，促进它们的正常分化。这个提议 试图了解这些共同因素如何激活不同的分化途径。第一个目标将 寻找视网膜分化过程中受体酪氨酸激酶信号传导和 Glass 的直接靶基因。 靶向DNA腺嘌呤甲基转移酶鉴定（DamID）将用于鉴定直接靶基因 Pointed 是介导表皮生长因子受体 (EGFR) 信号传导的转录因子， 未分化细胞、感光细胞和视锥细胞。类似的方法将用于识别 Glass 目标 基因，并且两个数据集将与改变其表达的基因的转录组分析进行比较 在 Egfr 或玻璃突变体中。这些实验的目的是鉴定诱导的转录因子 通过 EGFR 信号传导促进感光细胞和视锥细胞的分化，并阐明 Glass 如何 纳入他们的监管。第二个目标涉及转录抑制因子如何限制 玻璃驱动细胞类型特异性基因表达。已知可驱动的两个明确的监管区域 以玻璃依赖性方式的光感受器特异性表达将用于识别阻遏物 阻止 Glass 激活视锥细胞和色素细胞中的这些基因。这些阻遏物的重要性 结合位点将在内源基因组环境中进行测试。在补充方法中，增强剂 将识别并特别在视锥细胞或色素细胞中驱动玻璃依赖性表达的区域 他们的监管特点。结合起来，这些实验将揭示不同的细胞身份如何能够 使用一组有限的信号和转录因子从共同祖细胞中指定，这一过程 也发生在哺乳动物的视网膜中。结果将有助于提高我们生产特定细胞类型的能力 来自用于再生医学的干细胞。