Biased sweet taste signaling pathways in mice and humans

小鼠和人类中偏向的甜味信号通路

• 批准号: 10867691
• 负责人: Sunil Kumar Sukumaran
• 金额: $ 8.68万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10867691
• 关键词: 3D Print; ARRB1 gene; Ablation; Adaptor Signaling Protein; Arrestins; Attention; Behavioral; Binding; Biological Assay; Biological Models; Biopsy; Calories; Cells; Consumption; Data; Development; Disease; Endocytosis; Energy Transfer; Food; Fructose; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gene Expression; Gene Expression Profile; Genes; Genomics; Grant; Growth Factor; Health Benefit; Human; Immunohistochemistry; In Situ Hybridization; Industry; Inferior; Kinetics; Knockout Mice; Ligands; LoxP-flanked allele; MAPK3 gene; Measures; Mediating; Mediator; Microscopic; Mitogen-Activated Protein Kinases; Mus; Natural regeneration; Nebraska; Nerve; Obesity Epidemic; Organoids; Pathway interactions; Pharmacology; Phosphorylation; Physiological; Play; Positioning Attribute; Role; Saccharin; Signal Pathway; Signal Transduction; Stream; Sucrose; Sweetening Agents; Tamoxifen; Taste Buds; Taste Perception; Taste preferences; Testing; Transfection; United States National Institutes of Health; Western Blotting; cell type; conditional knockout; desensitization; design; dietary; drug candidate; experience; good diet; in vivo; model organism; mouse model; novel; novel strategies; obesity prevention; prevent; receptor; response; single-cell RNA sequencing; stem cells; sugar; sweet taste perception; sweetened beverage; taste stimuli; taste system; tongue papilla; tool development; transcriptome sequencing; transcriptomics

The obesity epidemic has reached alarming proportions in many parts of the world, fueled by the over consumption of calorie-rich foods, primarily sugars in the form of sugar-sweetened beverages and other sweet foods. Hence, novel strategies are needed to prevent excessive sugar consumption and to promote a healthy diet. Non-caloric sweeteners have been used for several decades to replace sugar-derived calories. However, their health benefits have been questioned and their taste profile is inferior to sugars. Therefore, there is a need to develop novel healthy and tasty non-caloric sweeteners. The sweet taste receptor - a heterodimeric G-protein coupled receptor (GPCR) formed by the TAS1R2 and TAS1R3 subunits - is the primary receptor for sugars and non-caloric sweeteners. Studies to date have focused on the G-protein mediated pathway as the primary mechanism for transduction of sweet taste signaling downstream of the sweet taste receptor. However, it is now well known that the arrestins, adaptor proteins first identified as mediators of GPCR- desensitization, can transduce signals down stream of GPCRs on their own. The G-protein and arrestin pathways engage different downstream signaling partners and consequently, have different physiological effects. An exciting development in pharmacology is the discovery that some GPCR ligands differentially engage the G-protein and arrestin pathways, although they bind to the same receptor. Such biased ligands are exciting drug candidates. Using single-cell RNASeq, we showed that Arrb1 is the sole arrestin expressed in sweet taste receptor expressing cells. We propose to identify the role of arrestin signaling in sweet taste using conditional knockout (CKO) mice models and taste organoids cultured from this strain. We will confirm the specific expression of Arrb1 in sweet taste cells using in situ hybridization and immunohistochemistry and will compare the behavioral and taste nerve responses of Arrb1CKO and control mice to sweet and other control taste stimuli. The kinetics of arrestin binding to the sweet taste receptor, sweet taste adaptation, and arrestin signaling will be studied in taste organoids using microscopic and other assays (Specific Aim 1). Unfortunately, such mechanistic studies are not possible in humans, as human taste stem cells have not been identified. We will use cutting-edge spatial transcriptomics of human and mouse taste papillae and compare this to existing droplet-based scRNASeq data from mouse cells, to identify human taste cell types including stem cells and the growth factors required for regeneration of taste cells (Specific Aim 2). Data from this study will enable the development of tools and strategies to manipulate sweet taste signaling.

肥胖流行在世界许多地方都达到了令人震惊的比例 消费富含卡路里的食物，主要是糖味的饮料和其他糖的形式 甜食。因此，需要采取新颖的策略来防止过量的糖消耗并促进 健康的饮食。非热甜味剂已使用数十年来代替糖衍生 卡路里。但是，他们的健康益处受到质疑，其口味特征不如糖。 因此，有必要开发出新颖的健康和美味的非热味甜味剂。甜味 受体 - 由TAS1R2和TAS1R3形成的异二聚体G蛋白偶联受体（GPCR） 亚基 - 是糖和非含量甜味剂的主要受体。迄今为止的研究重点是 G蛋白介导的途径是转导甜味信号传导的主要机制 甜味受体的下游。但是，现在众所周知，逮捕蛋白，适配器蛋白 首先被识别为GPCR脱敏的介体，可以在GPCR上传播信号 他们自己的。 G蛋白和逮捕途径与不同的下游信号合作伙伴与 因此，具有不同的生理影响。药理学方面的令人兴奋的发展是 发现某些GPCR配体有差异地与G蛋白和逮捕途径相关，尽管他们 与同一受体结合。这种偏见的配体是令人兴奋的候选药物。使用单细胞RNASEQ，我们 表明ARRB1是在甜味受体表达细胞中表达的唯一阻止素。我们建议 使用条件敲除（CKO）小鼠模型确定逮捕蛋白信号传导在甜味中的作用，并且 从这种菌株中培养的味类器官。我们将确认ARRB1在甜味中的特定表达 使用原位杂交和免疫组织化学的细胞，将比较行为和味觉神经 ARRB1CKO和控制小鼠对甜和其他对照味刺激的反应。逮捕的动力学 将研究与甜味受体的结合，甜味适应和阻止蛋白信号传导。 使用微观和其他测定法（特定目标1）的器官。不幸的是，这种机械研究 由于尚未确定人类味道干细胞，因此在人类中不可能。我们将使用尖端 人和老鼠的空间转录组学味道乳头，并将其与现有的基于液滴进行比较 来自小鼠细胞的SCRNASEQ数据，以鉴定包括干细胞在内的人味细胞类型和生长 味觉细胞再生所需的因素（特定目标2）。这项研究的数据将使 制定操纵甜味信号的工具和策略。