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R 03提案是根据标题为“试点项目”的RFA-RM-21-012编写的 研究未充分研究的G蛋白偶联受体、离子通道和蛋白激酶。我们的研究将 重点关注G蛋白偶联受体家族中的人苦味受体（TAS 2 Rs），其表达是 在呼吸系统中检测到。这些受体已被指定为开放研究的合格蛋白质， 与RFA相关的NIH的照亮可药用基因组（IDG）计划，以及它们在苦 需要阐明基于促味剂/TAS 2 R激动剂的哮喘疗法。哮喘是一种主要的公共卫生 挑战和儿科人群中最常见的慢性疾病。在美国，超过2500万 包括8.4%的儿童在内的200万人目前患有哮喘，估计每年的费用约为82美元 亿目前，主要的治疗策略是基于β2-肾上腺素能受体激动剂，皮质类固醇， 和单克隆抗体。然而，这些药物的长期疗效和安全性仍存在问题。 接近。此外，对于进行性气道重塑仍然没有有效的治疗方法，这在气道重塑中起着重要作用。 在哮喘相关死亡中的关键作用。所有这些因素都为苦味的研究提供了强大的动力 促味剂/TAS 2 R激动剂用于治疗哮喘，因为最近研究表明这些药物具有 在动物模型中治疗哮喘的上级功效，其中数据表明这些激动剂可能 克服与当前治疗相关的关键缺陷， 重塑我们努力的长期目标是进一步了解TAS 2 R的机制， 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 提出测试苦味受体在促苦味剂/TAS 2 R激动剂中起重要作用的假设- 哮喘的治疗策略。因此，我们将工程改造Tas 2 r基因缺陷的小鼠突变体， 或者通过使用高效的基于CRISPR的技术携带单个Tas 2 r基因。我们还将比较 促苦味剂/TAS 2 R激动剂治疗纯合子哮喘和相关信号传导的结果 突变体和野生型同窝仔。这些实验将帮助我们更好地了解分子 促苦味剂/TAS 2 R激动剂治疗哮喘的潜在机制。我们的研究还将揭示， TAS 2 R对小鼠的生存至关重要。最后，本研究期间开发的遗传资源将 TAS 2 R的生理、病理和治疗作用的有力工具， 医学状况，如肥胖症、糖尿病和早产。