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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：
10215441
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/10215441
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.

在世界许多地方，肥胖症的流行已经达到了令人担忧的程度，高热量食物的消费，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的糖消费，并促进健康的饮食无热量甜味剂已被用于几十年来取代糖衍生的甜味剂。卡路里然而，它们的健康益处受到质疑，它们的味道不如糖。因此，需要开发新的健康和美味的无热量甜味剂。甜味受体-由TAS 1 R2和TAS 1 R3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。迄今为止的研究集中在 G蛋白介导的途径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，抑制蛋白，衔接蛋白，首先被鉴定为GPCR脱敏的介质，可以将GPCR下游的信号传递到自己的. G蛋白和抑制蛋白通路参与不同的下游信号传导伙伴，因此具有不同的生理效应。药理学的一个令人兴奋的发展是发现一些GPCR配体差异地参与G蛋白和抑制蛋白途径，尽管它们与相同的受体结合。这种偏向配体是令人兴奋的候选药物。使用单细胞RNASeq，我们表明Arrb 1是甜味受体表达细胞中表达的唯一抑制蛋白。我们建议使用条件性敲除（CKO）小鼠模型鉴定arrestin信号传导在甜味中的作用，从这种菌株中培养的味觉类器官。我们将进一步证实Arrb 1在甜味中的特异性表达细胞采用原位杂交和免疫组织化学，并将比较行为和味觉神经 Arrb 1CKO和对照小鼠对甜味和其他对照味觉刺激的反应。抑制蛋白的动力学结合到甜味受体，甜味适应，和arrestin信号将在味觉研究使用显微镜和其他测定的类器官（特定目标1）。不幸的是，这种机械研究在人类中是不可能的，因为人类的味觉干细胞还没有被识别出来。我们将使用最先进的人和小鼠味觉乳头空间转录组学，并将其与现有的基于液滴的来自小鼠细胞的scRNASeq数据，以鉴定包括干细胞在内的人类味觉细胞类型，并在小鼠细胞中生长。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。