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Biased sweet taste signaling pathways in mice and humans

小鼠和人类中偏向的甜味信号通路

基本信息

批准号：
10456040
负责人：
Sunil Kumar Sukumaran
金额：
$ 13.26万
依托单位：
UNIVERSITY OF NEBRASKA LINCOLN
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-05 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456040
关键词：
3D Print ARRB1 gene Adaptor Signaling Protein Animal Model 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 Goals Grant Growth Factor Growth Factor Receptors Health Benefit Human Immunohistochemistry In Situ Hybridization Industry Inferior Kinetics Ligands MAPK3 gene Measures Mediating Mediator of activation protein Methods 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 Sugar Substitute Sweetening Agents Tamoxifen Taste Buds Taste Perception Taste preferences Testing Tissues United States National Institutes of Health Western Blotting base cell type conditional knockout desensitization design dietary drug candidate experience good diet in vivo mouse model novel novel strategies obesity prevention prevent receptor response single-cell RNA sequencing stem cell growth stem cells sugar sweet taste perception sweetened beverage taste stimuli taste system tongue papilla tool development transcriptome sequencing transcriptomics

项目摘要

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.

在过度肥胖的推动下，世界许多地区的肥胖流行已达到惊人的程度。食用高热量食物，主要是含糖饮料和其他形式的糖甜食。因此，需要新的策略来防止过量糖消费并促进健康的饮食。几十年来，无热量甜味剂一直用于替代糖源甜味剂卡路里。然而，它们的健康益处受到质疑，而且它们的味道不如糖。因此，需要开发新型健康美味的无热量甜味剂。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 - 是糖和无热量甜味剂的主要受体。迄今为止的研究主要集中在 G蛋白介导的途径作为甜味信号转导的主要机制甜味受体的下游。然而，现在众所周知，视紫红质抑制蛋白、接头蛋白首先被鉴定为 GPCR 脱敏介体，可以在 GPCR 下游转导信号他们自己的。 G 蛋白和视紫红质抑制蛋白途径涉及不同的下游信号传导伙伴，因此，具有不同的生理作用。药理学的一个令人兴奋的发展是发现一些 GPCR 配体差异性地参与 G 蛋白和抑制蛋白途径，尽管它们与相同的受体结合。这种有偏向的配体是令人兴奋的候选药物。使用单细胞 RNASeq，我们表明 Arrb1 是甜味受体表达细胞中唯一表达的抑制蛋白。我们建议使用条件敲除 (CKO) 小鼠模型确定抑制蛋白信号在甜味中的作用，并品尝从该菌株培养的类器官。我们将确认Arrb1在甜味中的具体表达使用原位杂交和免疫组织化学对细胞进行比较，并比较行为和味觉神经 Arrb1CKO 和对照小鼠对甜味和其他对照味觉刺激的反应。抑制蛋白的动力学将在味觉中研究与甜味受体的结合、甜味适应和抑制蛋白信号传导使用显微镜和其他检测方法检测类器官（具体目标 1）。不幸的是，这样的机制研究这在人类中是不可能的，因为人类味觉干细胞尚未被识别。我们将用最尖端的人类和小鼠味觉乳头的空间转录组学，并将其与现有的基于液滴的来自小鼠细胞的 scRNASeq 数据，用于识别人类味觉细胞类型，包括干细胞和生长味觉细胞再生所需的因子（具体目标 2）。这项研究的数据将使开发操纵甜味信号的工具和策略。