Function and origin of circumvallate taste bud cells from SOX10 expressing von Ebners glands

表达 SOX10 的冯·艾伯纳腺体周围味蕾细胞的功能和起源

• 批准号: 10374910
• 负责人: Hongxiang Liu
• 金额: $ 21.3万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10374910
• 关键词: Abbreviations; Adopted; Adult; Cell Communication; Cell Differentiation process; Cell physiology; Cells; Circumvallate Papilla; Clinical; Complex; Connective Tissue; Connective Tissue Cells; Data; Data Set; Derivation procedure; Development; Duct (organ) structure; Ductal Epithelial Cell; Eating; Embryo; Epithelial; Epithelial Cells; Esthesia; Eye; Foliate Papilla; Fungiform Papilla; Future; Gene Expression; Gland; Goals; Homeostasis; In Situ Hybridization; In Vitro; Individual; LGR5 gene; Label; Life; Location; Longevity; Mammals; Maps; Methods; Minor salivary gland structure; Mitotic; Modernization; Molecular; Mus; Neural Crest; Nutritional; Organ; Population; Property; Publishing; Quality of life; Radiation therapy; Regulation; Reporting; Rodent; Sensory; Sodium Chloride; Soft Palate; Solid; Source; Specialized Epithelial Cell; Stratum Basale; Structure; System; Taste Bud Cell; Taste Buds; Taste Disorders; Taste Perception; Techniques; Tissue Transplantation; Tissues; Tongue; Transplantation; Type III Epithelial Receptor Cell; cell type; chemotherapy; effective therapy; insight; migration; novel; postnatal; progenitor; single-cell RNA sequencing; stem cells; three dimensional cell culture; tongue papilla; tool; transcriptomics

Project Summary/Abstract (abbreviations: TB - taste bud; CvP – circumvallate papilla; vEG – von Ebner's gland) In mammals, the sensory end organs for taste are taste buds (TBs) that reside primarily in the epithelium of taste papillae in the tongue -- fungiform, foliate, and circumvallate. In rodents the single circumvallate papilla (CvP) houses hundreds of TBs, which enables efficient analyses of TB cells and will be our focus in this proposal. TB cells are post-mitotic and they have a short lifespan and undergo continuous renewal, therefore, progenitors in the immediate surrounding tissue must be available to renew TB cells for homeostasis. Thorough information of TB cell function, origin, and differentiation is essential to understanding taste disorders caused by TB progenitor deficiencies. TB progenitor cells have been thought to reside in the basal layer of the stratified lingual epithelium and express Krt14, Krt5, p63, SOX2 and Lgr5. However, our recent published data have shown that certain TB cells (~35% of TB cells in the CvP) originate from a previously unappreciated source of progenitors under, but not within, the stratified tongue epithelium -- the SOX10-expressing (SOX10+) cells. The labeled TB cells were mainly type III that transduce a sour and a salt taste, and had a significantly lower level of pan-taste cell marker Krt8. Further studies on the tissue locations and lineage mapping of SOX10+ cells have led to the idea that von Ebner's gland (vEG) is the tissue nest of SOX10+ progenitors for TB cells in the CvP. This is supported by the report that TBs-containing “circular structures” formed where vEG ducts locate in transplanted CvP tissue. The goal of this application is to collect direct and solid evidence about whether SOX10+ ductal cells of vEGs migrate and differentiate to TB cells (Aim 1), and establish the transcriptomic profiles of CvP/vEG epithelial cells, including SOX10+ progenitors and their derived TB cells (Aim 2). In Aim 1, we will adopt the previously proven approaches for examining taste cell differentiation - the 3D cultures of dissociated cells and CvP/vEG transplantations. In addition to taking advantage of these in vitro and ex vivo systems, we will utilize genetically labeled (RFP+) SOX10+ ductal cells of vEGs and CvP/vEG complex from SOX10-iCreERT2/RFP mice to differentiate taste cells in 3D cultures and transplanted tissues. In Aim 2, we will take advantage of the power of a modern high-throughput technique, single cell (sc) RNA-Seq that enables a highly efficient and informative analysis, to establish the transcriptomic profiles of all types of CvP/vEG epithelial cells including SOX10+ progenitors in vEGs and derived TB cells. The yielded data will provide definitive information about the differentiation of SOX10+ vEG cells to TB cells and bring comprehensive information of gene expression for all CvP TB cells and surrounding epithelial cells including those TB progenitors such as SOX10+ vEG ductal cells. The data will be of significant value for a long term in facilitating studies for a clear understanding of TB cell functions and the mechanisms, tissue compartments and cell types of TB progenitors in CvP/vEGs. Key words: taste bud (TB); von Ebner's gland (vEG); circumvallate papilla (CvP); SOX10; progenitor.

项目总结/摘要（缩写：TB - 味蕾；CvP - 环乳头；vEG - 冯·埃布纳腺） 在哺乳动物中，味觉末端器官是味蕾（TB），主要存在于味觉上皮细胞中。 味觉舌头上的乳头——菌状、叶状和周状。在啮齿类动物中，单个外周乳头 (CvP) 容纳了数百个 TB，可以对 TB 细胞进行高效分析，这将是我们在此方面的重点 提议。结核细胞是有丝分裂后的细胞，它们的寿命很短，并且会不断更新，因此， 周围组织中的祖细胞必须能够更新结核细胞以实现体内平衡。 有关结核细胞功能、起源和分化的全面信息对于了解味觉障碍至关重要 由结核祖细胞缺陷引起。 结核祖细胞被认为存在于复层舌上皮的基底层中 表达 Krt14、Krt5、p63、SOX2 和 Lgr5。然而，我们最近发布的数据表明，某些结核病 细胞（CvP 中约 35% 的 TB 细胞）源自以前未受重视的祖细胞来源，但 不在其中的是复层舌上皮——表达 SOX10（SOX10+）的细胞。标记的结核细胞是 主要是III型，转导酸味和咸味，并且泛味细胞标记物水平显着较低 克尔特8。对 SOX10+ 细胞的组织位置和谱系图谱的进一步研究得出了这样的想法：von 艾伯纳腺 (vEG) 是 CvP 中 TB 细胞 SOX10+ 祖细胞的组织巢。这是由 报道称，移植的 CvP 组织中 vEG 导管所在的位置形成了含有 TB 的“圆形结构”。 本申请的目的是收集关于 SOX10+ 导管细胞是否 vEG 迁移并分化为 TB 细胞（目标 1），并建立 CvP/vEG 的转录组谱 上皮细胞，包括 SOX10+ 祖细胞及其衍生的 TB 细胞（目标 2）。在目标 1 中，我们将采用 先前经过验证的检查味觉细胞分化的方法 - 分离细胞的 3D 培养物和 CvP/vEG 移植。除了利用这些体外和离体系统之外，我们还将利用 来自 SOX10-iCreERT2/RFP 的 vEG 和 CvP/vEG 复合物的基因标记 (RFP+) SOX10+ 导管细胞 小鼠在 3D 培养物和移植组织中区分味觉细胞。在目标 2 中，我们将利用 现代高通量技术单细胞 (sc) RNA-Seq 的强大功能可实现高效且 信息分析，建立所有类型的 CvP/vEG 上皮细胞的转录组谱，包括 vEG 和衍生结核细胞中的 SOX10+ 祖细胞。 产生的数据将提供有关 SOX10+ vEG 细胞分化为 TB 细胞的明确信息 并带来所有CvP TB细胞和周围上皮细胞基因表达的全面信息 包括那些结核祖细胞，例如 SOX10+ vEG 导管细胞。这些数据将在很长一段时间内具有重要价值 促进研究的术语，以清楚地了解结核细胞的功能和机制、组织 CvP/vEG 中结核祖细胞的区室和细胞类型。 关键词：味蕾（TB）；冯·埃布纳氏腺（vEG）；外周乳头 (CvP)； SOX10；祖。