Asthma remains a major public health issue worldwide, affecting all ages and ethnicities, with 50% of patients experiencing inadequate control of the disease. Constricted airways from contraction of airway smooth muscle is the major cause of airflow obstruction, and morbidity and mortality, in asthma. Despite this key point for intervention, only one class of direct bronchodilators (?-agonists) are available for treatment, and they are suboptimal for many patients. We have discovered bitter taste receptors (TAS2Rs) on human airway smooth muscle (HASM) cells which signal by a unique mechanism. When activated, TAS2Rs markedly relax HASM and relieve airway obstruction that is otherwise resistant to ?-agonists. However, the known TAS2R agonists have low affinity and evoke desensitization (tachyphylaxis) of the relaxation response over time via ?-arrestin mechanisms. The overarching hypothesis is that TAS2R agonists with novel structures can be highly effective in relaxing airways in an asthmatic milieu and yet not evoke tachyphylaxis. The broad long-term objectives are to determine the 3D structures and binding sites of two TAS2Rs expressed on HASM (R5 and R14) and then use these structures to perform virtual docking of an agnostic ultra-large compound library to identify potential ligands. These hits will be examined in engineered model cells, but will be most intensely studied within the context of the physiology of HASM cells, and human airways, under asthmatic conditions to delineate novel ways to engage the receptor that are biased towards relaxation and away from desensitization.
Aim 1 will computationally determine the structures and binding sites for R5 and R14 using methods that include 13 trillion combinations of the residues to determine favorable energy conformations of inactive and active states. The structures will be used to dock compounds from a highly diverse library. These will be studied in Aim 2 to determine potency and efficacy in genetically engineered model cells and in HASM cells from nonasthmatic and asthmatic lungs.
In Aim 3, the most favorable compounds will be studied to ascertain ?-arrestin engagement and biasing away from desensitization in model cells, HASM cells, and human lungs under asthmatic conditions. Results from Aims2/3 will be fed back into Aim1 to further refine a model of biased agonists for these HASM TAS2Rs. Based on our preliminary data that support all three Aims, we will learn about the structural requirements for biasing for these receptors. And, we anticipate that multiple highly effective agonists with unexpected structures will be identified and that their bronchodilating properties in asthma will represent a new class of powerful non-desensitizing agents for treating and preventing bronchospasm.
Many asthmatics do not have adequate control over air flow due to airway obstruction, and treatment options are limited to one class of bronchodilator drugs which all work the same way. The proposal seeks to determine the structure of a different class of bronchodilator drug acting at bitter taste receptors on airway smooth muscle, and then find drugs with favorable pharmacologic characteristics to activate these receptors. This will lead to increased treatment options for asthma that could decrease patient morbidity and mortality.
