Elucidating the role of Polycomb Repressive Complexes in Lingual Papillae Development

阐明多梳抑制复合物在舌乳头发育中的作用

• 批准号: 10200755
• 负责人: Elena Ezhkova
• 金额: $ 35.83万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10200755
• 关键词: Address; Anterior; Antibodies; Basal Cell; Binding; Biological Assay; Candidate Disease Gene; Cell Differentiation process; Cell Extracts; Cell Lineage; Cell Maintenance; Cells; ChIP-seq; Chromatin; Communication; Complex; Connective Tissue; Coupled; Data; Defect; Development; Drosophila genus; Eating; Embryo; Embryonic Development; Epithelial; Epithelial Cells; Esthesia; Failure; Filiform Papilla; Fungiform Papilla; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Histone H2A; In Vitro; Ligands; Maintenance; Mastication; Mediating; Mesenchymal; Microglossias; Molecular; Morphogenesis; Mus; Muscle; Ontology; Organ; PRC1 Protein; Pathway interactions; Pattern; Pattern Formation; Phenotype; Play; Polycomb; Process; Protocols documentation; RNA; Regulation; Research Proposals; Role; SHH gene; Side; Signal Pathway; Structure; Taste Buds; Taste Perception; Testing; Tissues; To specify; Tongue; Transcription Repressor; Transcriptional Regulation; WNT Signaling Pathway; appendage; base; chromatin immunoprecipitation; embryonic stem cell; experimental study; gene repression; in vivo; insight; keratinization; loss of function; novel; oral tissue; progenitor; recruit; smoothened signaling pathway; stem cells; tongue papilla; transcription factor; transcriptome sequencing

Summary The tongue is a complex organ that is essential for mastication, taste sensation, and communication. Proper development of the tongue is dependent on complex interactions between different tissues, including muscle, connective tissue, and epithelium. Importantly, failure to establish these interactions leads to developmental defects, such as aglossia, microglossia, and ankyloglossia. The lingual epithelium is an array of epithelial-mesenchymal appendages termed papillae. Fungiform papillae harbor taste buds required for taste sensation. They are surrounded by keratinized, non-gustatory filiform papillae, which provide essential barrier functions. Wnt and Shh signaling pathways are central players in regulating the patterning and development of fungiform and filiform papillae; however, the role of cell-intrinsic mechanisms in the specification and maintenance of these structures remains unclear. Polycomb repressive complex (PRC) 1 is a major chromatin transcriptional regulator and has been shown to play a critical role in lineage identity and early embryonic development; however, PRC1-mediated regulation of cell lineages has yet to be explored in oral tissues. To investigate the role of PRC1 in the development of lingual papillae, we generated mice in which essential PRC1 subunits, Ring1a and Ring1b, were conditionally ablated in the basal lingual epithelial cells (Ring1a/b 2KO). Our results showed that in Ring1a/b 2KO mice, the fungiform papillae are not maintained and the filiform papillae are not formed. Further analysis of the Ring1a/b-null lingual epithelium revealed widespread expression of the Sonic Hedgehog (Shh) ligand and the transcription factor Sox2, whereas in control mice, the expression of these genes was confined to taste progenitors. The expression pattern of components of the Wnt signaling pathway, Wnt10b and Lef1, was also altered in Ring1a/b 2KO; however, this did not result in aberrant canonical Wnt signaling. We hypothesize that PRC1 restricts gene expression and thus promotes lingual papilla patterning and formation. To dissect the molecular mechanisms of PRC1's control of the lingual epithelium, we will carry out an RNA-seq analysis of FACS-purified control and Ring1a/b- null lingual epithelial cells. This analysis will be coupled with ChIP-seq with antibodies against PRC1 subunits to identify the direct targets of PRC1. To identify the specific molecular pathways disrupted in the Ring1a/b-null lingual epithelium, we will carry out Gene Ontology analysis and focus on candidate genes that regulate epithelial-mesenchymal interactions, transcriptional regulation, and lineage identity. We will analyze the functional significance of the candidate genes by performing genetic experiments in vivo. Together, our studies will allow us to elucidate PRC1's regulation of the lingual epithelium, including its interaction with developmental signaling pathways. Understanding the complex molecular interactions between lingual tissues may also help us to reveal the mechanisms that cause congenital tongue abnormalities to form.

概括 舌头是一个复杂的器官，对于咀嚼、味觉和交流至关重要。 舌头的正常发育取决于不同组织之间复杂的相互作用，包括 肌肉、结缔组织和上皮。重要的是，未能建立这些相互作用会导致 发育缺陷，例如失语、小舌和舌强直。舌上皮是一系列 上皮间质附属物称为乳头。菌状乳头含有味觉所需的味蕾 感觉。它们被角化的、非味觉的丝状乳头包围，这些乳头提供了必要的屏障 功能。 Wnt 和 Shh 信号通路是调节细胞模式和发育的核心参与者。 菌状和丝状乳头；然而，细胞内在机制在规范和 这些结构的维护仍不清楚。 Polycomb 抑制复合物 (PRC) 1 是主要染色质 转录调节因子，已被证明在谱系识别和早期胚胎发育中发挥关键作用 发展;然而，PRC1 介导的细胞谱系调节尚未在口腔组织中进行探索。 为了研究 PRC1 在舌乳头发育中的作用，我们培育了小鼠，其中 必要的 PRC1 亚基 Ring1a 和 Ring1b 在基底舌上皮细胞中被条件性消融 （环1a/b 2KO）。我们的结果表明，在 Ring1a/b 2KO 小鼠中，菌状乳头没有得到维持，并且 丝状乳头未形成。对 Ring1a/b-null 舌上皮的进一步分析揭示 Sonic Hedgehog (Shh) 配体和转录因子 Sox2 的广泛表达，而 对照小鼠中，这些基因的表达仅限于味觉祖细胞。的表达模式为 Wnt 信号通路的组成部分 Wnt10b 和 Lef1 在 Ring1a/b 2KO 中也发生了改变；然而，这 没有导致异常的规范 Wnt 信号传导。我们假设 PRC1 限制基因表达并且 从而促进舌乳头的形成和形成。剖析 PRC1 的分子机制 为了控制舌上皮细胞，我们将对 FACS 纯化的对照和 Ring1a/b-进行 RNA 序列分析 舌上皮细胞无效。该分析将与带有 PRC1 亚基抗体的 ChIP-seq 相结合 确定 PRC1 的直接目标。鉴定 Ring1a/b-null 中被破坏的特定分子途径 舌上皮细胞，我们将进行Gene Ontology分析，重点寻找调控的候选基因 上皮-间质相互作用、转录调控和谱系同一性。我们将分析 通过进行体内基因实验来确定候选基因的功能意义。 总之，我们的研究将使我们能够阐明 PRC1 对舌上皮的调节，包括其 与发育信号通路的相互作用。了解之间复杂的分子相互作用 舌组织还可以帮助我们揭示导致先天性舌头异常形成的机制。