CRISPR-mediated engineering and pilot study of mouse mutants of the bitter taste receptor genes

CRISPR介导的小鼠苦味受体基因突变体工程和初步研究

• 批准号: 10451169
• 负责人: Eugene Yu
• 金额: $ 18.23万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-17 至 2025-05-16
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10451169
• 关键词: Address; Adrenal Cortex Hormones; Adrenergic Agonists; Agonist; Alleles; Allergens; Animal Model; Area; Asthma; Biological; Biological Process; Bronchodilation; CRISPR/Cas technology; Cause of Death; Cell Proliferation; Cells; Cessation of life; Child; Childhood; Chromosomes; Chronic Disease; Clinical; Clinical Management; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Cost of Illness; Data; Diabetes Mellitus; Disease; Drug Targeting; Engineering; Family; Family member; Financial compensation; Funding; G-Protein-Coupled Receptors; Gene Cluster; Gene Family; Genes; Genome; Goals; Health Care Costs; Human; Individual; Inflammatory Response; Investigation; Ion Channel Protein; Mediating; Medical; Molecular; Monoclonal Antibodies; Mus; Muscarinic Acetylcholine Receptor; Muscle relaxation phase; Mutant Strains Mice; Obesity; Obstructive Lung Diseases; Oral cavity; Orthologous Gene; Outcome; Pathologic; Pathologic Processes; Persons; Pharmacology; Phenotype; Physiological; Pilot Projects; Play; Population; Premature Labor; Process; Protein Kinase; Proteins; Public Health; Quinine; Receptor Gene; Refractory; Regulation; Research; Resources; Respiratory System; Role; Safety; Signal Transduction; Smooth Muscle Myocytes; Taste Buds; Technology; Testing; Therapeutic; Therapeutic Effect; Therapeutic Studies; Treatment outcome; United States; United States National Institutes of Health; airway remodeling; allergic airway inflammation; antagonist; base; beta-2 Adrenergic Receptors; body system; cost; desensitization; druggable target; effective therapy; experimental study; genetic resource; genetic testing; insight; mast cell; mouse model; mutant; novel; prevent; programs; receptor; respiratory smooth muscle; response; success; tool; treatment strategy

ABSTRACT: This R03 proposal has been prepared in response to RFA-RM-21-012 entitled “Pilot Projects Investigating Understudied G Protein-Coupled Receptors, Ion Channels, and Protein Kinases.” Our research will focus on human bitter taste receptors (TAS2Rs) in the G protein-coupled receptor family, whose expression is detected in the respiratory system. These receptors have been designated as eligible proteins open for study by NIH’s Illuminating the Druggable Genome (IDG) Program associated with this RFA, and their roles in bitter tastant/TAS2R-agonist-based therapies for asthma need to be elucidated. Asthma is a major public health challenge and the most common chronic disease in the pediatric population. In the United States, over 25 million people, including 8.4% of all children, are currently suffering from asthma with an estimated annual cost of ~$82 billion. At present, the main treatment strategies are based on β2-adrenergic receptor agonists, corticosteroids, and monoclonal antibodies. However, questions remain about the long-term efficacy and safety of these approaches. In addition, there is still no effective treatment for progressive airway remodeling, which plays a critical role in asthma-related deaths. All these factors provide a strong impetus for investigations of bitter tastants/TAS2R agonists for treating asthma because it has been recently shown that these agents have a superior efficacy for asthma treatment in animal models, in which data suggest that these agonists can potentially overcome critical deficiencies associated with current therapeutics such as those related to progressive airway remodeling. The long-term goal of our efforts is to further understand TAS2Rs mechanistically in terms of the physiological, pathological, and therapeutic processes associated with asthma treatment. In this pilot project, we propose to test the hypothesis that bitter taste receptors play an essential role in bitter tastant/TAS2R-agonist- based therapeutic strategies for asthma. Accordingly, we will engineer mouse mutants deficient for Tas2r genes or carrying a single Tas2r gene by using highly efficient CRISPR-based technology. We will also compare the outcomes of bitter tastants/TAS2R-agonist-based treatment of asthma and associated signaling in homozygous mutants and wild-type littermates. These experiments will help us to better understand the molecular mechanisms underlying bitter tastant/TAS2R-agonist-based therapies for asthma. Our study will also reveal if TAS2Rs are essential for the survival of mice. Lastly, the genetic resources developed during this study will be powerful tools for unraveling the physiological, pathological, and therapeutic roles of TAS2Rs related to other medical conditions, such as obesity, diabetes, and preterm labor.

摘要：该R03提案是针对RFA-RM-21-012的题为“试点项目”准备的 研究了正在研究的G蛋白偶联受体，离子通道和蛋白激酶。“我们的研究将会 专注于G蛋白偶联受体家族中的人类苦味受体（TAS2R），其表达是 在呼吸系统中检测到。这些受体已被指定为合格的蛋白质，可以通过 NIH的启发性基因组（IDG）计划与此RFA相关的计划及其在苦涩中的角色 需要阐明哮喘的味道/TAS2R激动剂疗法。哮喘是主要的公共卫生 挑战和小儿人群中最常见的慢性疾病。在美国，超过2500万 包括所有儿童在内的8.4％的人目前患有哮喘，估计年费用约为82美元 十亿。目前，主要治疗策略基于β2-肾上腺素能受体激动剂，皮质类固醇，皮质类固醇， 和单克隆抗体。但是，关于这些的长期效率和安全的问题仍然存在 方法。此外，仍然没有有效的渐进气道重塑治疗方法 在与哮喘相关的死亡中的关键作用。所有这些因素为痛苦的投资提供了强大的动力 品尝者/tas2r激动剂用于治疗哮喘，因为最近已经表明这些药物具有 动物模型中哮喘治疗的效率较高，其中数据表明这些激动剂可以潜在地 克服与当前疗法相关的关键缺陷，例如与渐进气道相关的疗法 重塑。我们努力的长期目标是从机械上进一步理解TAS2RS 与哮喘治疗相关的生理，病理和治疗过程。在这个试点项目中，我们 提议检验苦味受体在苦味/tas2r-agenist- 基于哮喘的治疗策略。彼此之间，我们将设计缺乏TAS2R基因的小鼠突变体 或通过使用高效的基于CRISPR的技术携带单个TAS2R基因。我们还将比较 基于肥胖的苦味和基于TAS2R激动剂的哮喘的治疗和相关信号在纯合子中 突变体和野生型窝窝。这些实验将帮助我们更好地理解分子 哮喘的基于苦味/TAS2R激动剂的苦味/TAS2R激动剂疗法的机制。我们的研究还将揭示是否是否 TAS2R对于小鼠的存活至关重要。最后，这项研究中开发的遗传资源将是 阐明与其他TAS2R相关的物理，病理和治疗作用的强大工具 肥胖，糖尿病和早产等医学状况。