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: 10218925
• 负责人: Hongxiang Liu
• 金额: $ 17.75万
• 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 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218925
• 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.

项目摘要/摘要（缩写：TB-味蕾； CVP - 绕过乳头; VEG - VEG - von Ebner的腺体） 在哺乳动物中，味觉的感觉端器官是味蕾（TBS），主要存在于 在舌头上品尝乳头 - 真菌状，叶状和环绕。在啮齿动物中 （CVP）容纳数百个TB，这可以有效地分析结核细胞，并将成为我们的重点 提议。 TB细胞是有丝分裂后的，它们的寿命很短，因此经历了连续的更新，因此 必须可以使用周围组织中的祖细胞来更新TB细胞以进行体内平衡。 TB细胞功能，起源和差异化的详尽信息对于理解味觉障碍至关重要 由结核病祖细胞缺陷引起。 TB祖细胞被认为存在于分层舌上上皮的基本层和 Express KRT14，KRT5，P63，SOX2和LGR5。但是，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）起源于先前未欣赏的祖细胞来源，但 不在内部，分层的舌上上皮 - 表达SOX10+的细胞。标记的TB细胞是 主要是转化酸味和盐味的III型，并且泛质细胞标记的水平明显较低 KRT8。关于Sox10+细胞的组织位置和谱系映射的进一步研究导致了Von的想法 Ebner的腺体（VEG）是CVP中TB细胞的Sox10+祖细胞的组织巢。这是由 报告说，含TBS的“圆形结构”形成，其中蔬菜管位于移植的CVP组织中。 该应用的目的是收集有关Sox10+导管细胞的直接证据 VEGS迁移并区分为TB细胞（AIM 1），并建立CVP/VEG的转录组轮廓 上皮细胞，包括Sox10+祖细胞及其衍生的TB细胞（AIM 2）。在AIM 1中，我们将采用 先前证明的用于检查味觉细胞分化的方法 - 解离细胞的3D培养物和 CVP/VEG移植。除了利用这些体外和离体系统外，我们还将利用 从SOX10-ICREERT2/RFP的CVP/VEG复合物的一般标记（RFP+）Sox10+导管细胞 小鼠在3D培养物和移植组织中区分味道细胞。在AIM 2中，我们将利用 现代高通量技术的力量，单细胞（SC）RNA序列，可实现高效且高效 信息分析，以建立所有类型的CVP/VEG上皮细胞的转录组曲线 蔬菜和衍生的TB细胞中的Sox10+祖细胞。 屈服的数据将提供有关Sox10+ Veg细胞与TB细胞区分的明确信息 并为所有CVP TB细胞和周围的上皮细胞带来全面的基因表达信息 包括那些结核病祖细胞，例如Sox10+ Veg导管细胞。长时间的数据将具有重要的价值 促进研究的术语，以清楚了解TB细胞功能和组织机制 CVP/VEGS中结核祖细胞的隔室和细胞类型。 关键词：味蕾（TB）；冯·埃伯纳（Von Ebner）的腺体（VEG）；围绕乳头（CVP）; Sox10;祖先。