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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：
10192528
负责人：
Sunil Kumar Sukumaran
金额：
$ 15.89万
依托单位：
UNIVERSITY OF NEBRASKA LINCOLN
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10192528
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 Diet Disease Endocytosis Energy Transfer Expression Profiling 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 drug candidate experience good diet in vivo 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

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.

肥胖症的流行在世界许多地区已经达到了令人震惊的程度，这是由摄入高卡路里食物，主要是含糖饮料和其他形式的糖甜食。因此，需要新的战略来防止过量的糖类消费并促进健康的饮食。几十年来，无热量甜味剂一直被用来取代糖类卡路里。然而，它们对健康的益处一直受到质疑，它们的口味特征不如糖。因此，有必要开发新型健康可口的无热量甜味剂。甜蜜的味道受体--Tas1r2和Tas1r3形成的异源二聚体G蛋白偶联受体亚单位-是糖和非卡路里甜味剂的主要受体。到目前为止的研究主要集中在 G蛋白介导的通路是甜味信号转导的主要机制甜味感受器的下游。然而，现在众所周知的是，拦阻蛋白，接头蛋白首先被确定为GPCR脱敏的媒介，可以将信号传导到GPCRs的下游他们自己的。G蛋白和arrestin通路参与不同的下游信号伙伴和因此，会产生不同的生理效应。药理学中一个令人兴奋的发展是发现一些GPCR配体不同地参与G蛋白和arrestin途径，尽管它们与相同的受体结合。这种有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是甜味受体表达细胞中唯一表达的arrestin。我们建议利用条件性基因敲除(CKO)小鼠模型研究arrestin在甜味中的作用品尝从该菌株培养出来的有机化合物。我们将确认Arrb1在甜味中的特异性表达细胞使用原位杂交和免疫组织化学，并将比较行为神经和味觉神经 Arrb1CKO和对照组小鼠对甜味和其他控制性味觉刺激的反应。芳香化反应动力学研究与甜味受体的结合、甜味适应和arrestin信号将在味觉中进行研究。使用显微镜和其他化验的有机物(具体目标1)。不幸的是，这种机械论的研究在人类身上是不可能的，因为人类的味觉干细胞还没有被鉴定出来。我们将使用尖端技术人类和小鼠味觉乳头的空间转录并与现有的基于液滴的比较来自小鼠细胞的scRNASeq数据，用于鉴定人类味觉细胞的类型，包括干细胞和生长情况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开发控制甜味信号的工具和策略。