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：本R03提案是为响应RFA-RM-21-012“试点项目”而准备的