Salivary gland ionocyte organization and function during homeostasis, repair, and disease

稳态、修复和疾病期间唾液腺离子细胞的组织和功能

基本信息

批准号：
10471333
负责人：
Helen P. Makarenkova
金额：
$ 58.15万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-18 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10471333
关键词：
Acinar Cell Acinus organ component Adult Affect Anions Biology Cells Characteristics Chronic Clinical Research Collaborations Complex Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Disease Disease Progression Duct (organ) structure Ductal Epithelial Cell Ductal Epithelium Electrolytes Enzymes Epithelial Epithelial Cells Exocrine Glands FGF10 gene Fibroblast Growth Factor Fluids and Secretions Fluorescence-Activated Cell Sorting Functional disorder Future Gene Expression Genes Gland Heterogeneity Homeostasis Immunoglobulins Inflammation Injury Intercalated Duct Ion Transport Ions Lead Maintenance Mesenchymal Mesenchyme Modeling Mucins Mus Myoepithelial cell Natural regeneration Parotid Gland Play Population Process Proteins Proton-Translocating ATPases RNA analysis Regulation Research Role Saliva Salivary Salivary Glands Salivary duct structure Signal Transduction Sjogren&apos s Syndrome Tamoxifen Testing Tetanus Helper Peptide Thrombospondin 1 Tissues Tomatoes Transplantation UXT gene cell type experimental study fibroblast growth factor receptor 2b gland development human disease human tissue improved functioning injured mouse model novel oral tissue overexpression postnatal postnatal development repaired single cell analysis single-cell RNA sequencing stem cell growth transcriptome sequencing vacuolar H+-ATPase

项目摘要

The salivary glands are exocrine glands that produce saliva. An adequate supply of saliva is critical to the maintenance of oral tissue. Saliva is composed of a variety of electrolytes and also contains immunoglobulins, proteins, enzymes and mucins, suggesting that saliva has a complex function. An important function of salivary gland ducts is to regulate ion homeostasis[1]. Dysfunction of epithelial ion homeostasis can lead to several human diseases[2, 3]. Our lineage tracing experiments using a tamoxifen inducible FGF10CreERT2:R26- Tomato mouse identified new epithelial populations of the Fgf10 expressing cells in the striated (SD), granular (GCT) and excretory ducts (ED) of submandibular (SMG) and parotid (PG) glands. Importantly Fgf10 lineage tracing shows that until postnatal day 7 (P7) only mesenchymal Fgf10 expressing cells are present, however after P10, Fgf10 expression in the mesenchyme shuts down, while a subset of ductal epithelial cells starts to express Fgf10. Using Fluorescence Activated Cell Sorting (FACS) we isolated both mesenchymal and epithelial populations of Fgf10+ cells and performed RNA sequencing (RNA-seq). Analysis of this RNA- seq data combined with analysis of single cell RNA-Seq (scRNAseq) data from P1 and adult SMGs showed that epithelial Fgf10+ populations overlap with the known duct population marked by Ascl3. Moreover, these epithelial Fgf10+ cells expressed many gene characteristics for an ancient type of ionocyte cell: Bsnd, Foxi1, Foxi2, Asgr1, Stap1, several subunits of the H+- ATPase (V-ATPase) and markers of SLC12A1/2 and Slc9a4 transporters. Most importantly Fgf10+ cells expressed cystic fibrosis transmembrane conductance regulator (Cftr) that plays a key role in exocrine secretion, including in salivary glands. We propose that these epithelial Fgf10+/Ascl3+/Cftr+ cells are specialized duct salivary gland ionocytes. Moreover, using two models of Sjogrens syndrome: the NOD.B10Sn-H2b/J and the thrombospondin-1 null (TSP1-/-) mice, we showed a significant decrease in Fgf10+ expression and the number of ionocytes with disease progression. We also showed that epithelial Fgf10+ cells isolated from the SMG are able to maintain proliferation and growth of progenitor and myoepithelial cells, suggesting an additional role for ionocytes in ductal maintenance. In this application we will determine the role of Fgf10 signaling in SMG regeneration and ionocyte function, define factors that control ionocyte differentiation and determine how chronic inflammation affects ionocyte function. In addition, in collaboration with Dr. Hoffman's group using bulk RNA-seq and scRNA-seq data we will investigate whether salivary glands have one type or several types of ionocytes. Our proposed study will provide a starting platform for future studies of ionocytes in basic biology and clinical research.

唾液腺是产生唾液的外分泌腺。供应充足唾液对于口腔组织的维护至关重要。唾液由多种电解质组成还含有免疫球蛋白、蛋白质、酶和粘蛋白，这表明唾液具有复杂的功能。唾液腺管的一个重要功能是调节离子体内平衡[1]。上皮离子稳态功能障碍可导致多种人类疾病疾病[2, 3]。我们使用他莫昔芬诱导型 FGF10CreERT2:R26- 进行谱系追踪实验番茄小鼠在横纹肌中鉴定出表达 Fgf10 的新上皮细胞群颌下管 (SMG) 和腮腺 (PG) 的 (SD)、颗粒管 (GCT) 和排泄管 (ED) 腺体。重要的是，Fgf10 谱系追踪显示，直到出生后第 7 天（P7）为止间充质 Fgf10 表达细胞存在，但是在 P10 后，Fgf10 表达在间充质关闭，而导管上皮细胞的一部分开始表达 Fgf10。使用荧光激活细胞分选 (FACS) 我们分离了间充质和上皮细胞 Fgf10+ 细胞群并进行 RNA 测序 (RNA-seq)。对该 RNA 的分析- seq 数据与 P1 和成体的单细胞 RNA-Seq (scRNAseq) 数据分析相结合 SMG 显示上皮 Fgf10+ 群体与标记的已知导管群体重叠通过 Ascl3。此外，这些上皮 Fgf10+ 细胞表达了许多基因特征古老类型的离子细胞：Bsnd、Foxi1、Foxi2、Asgr1、Stap1、H+- 的几个亚基 ATP 酶 (V-ATP 酶) 以及 SLC12A1/2 和 Slc9a4 转运蛋白的标记。最重要的是 Fgf10+ 细胞表达囊性纤维化跨膜电导调节因子 (Cftr) 在外分泌分泌中发挥关键作用，包括在唾液腺中。我们建议这些上皮 Fgf10+/Ascl3+/Cftr+ 细胞是专门的导管唾液腺离子细胞。此外，使用两个干燥综合征模型：NOD.B10Sn-H2b/J 和血小板反应蛋白-1 无效 (TSP1-/-) 小鼠中，我们发现 Fgf10+ 表达和离子细胞数量显着减少疾病进展。我们还表明，从 SMG 中分离出的上皮 Fgf10+ 细胞是能够维持祖细胞和肌上皮细胞的增殖和生长，这表明离子细胞在导管维护中的额外作用。在此应用程序中，我们将确定角色 Fgf10 信号在 SMG 再生和离子细胞功能中的作用，定义控制因素离子细胞分化并确定慢性炎症如何影响离子细胞功能。在此外，我们与 Hoffman 博士的团队合作，利用 Bulk RNA-seq 和 scRNA-seq 数据将研究唾液腺是否具有一种或多种类型的离子细胞。我们的拟议的研究将为基础生物学中离子细胞的未来研究提供一个起始平台和临床研究。