The beta2-adrenergic receptors (B2-ARs) are central in the regulation of the cardiovascular system and alterations in their signaling properties have been implicated in cardiovascular pathologies including hypertension and heart failure. The B2-ARs are also a main target of cardiovascular therapeutics- beta-blockers are commonly used in the treatment of heart failure. Elucidating the molecular mechanisms that control beta2-adrenoceptor signaling and mediate its functional consequences is therefore key to accomplishing the AHA objective, because it will illuminate how cells execute critical functions underlying cardiovascular physiology and will enhance our ability to develop more specific and efficient therapies for cardiovascular disorders. The B2-AR gene is highly polymorphic in the human population. Studies that have focused on a handful of common polymorphisms support their clinical and therapeutic significance, and reinforce that a comprehensive functional classification of all receptor variants will have critical impacts on disease diagnosis and effective treatment. However, advances in the functional classification of gene variants have been hindered by the lack of scalable assays to assess the phenotypes associated with large numbers of variants. This project aims to address this critical barrier by developing and applying an innovative high-throughput assay to systematically examine the functional impacts of all possible adrenoceptor gene variants. We will leverage cutting-edge CRISPR-based editing to generate all possible single nucleotide variants in the endogenous B2-AR locus. Then, we will combine a novel transcriptional reporter with next-generation sequencing to carry out a massively parallel analysis of the effects of genetic variants on receptor signaling in response to a panel of clinically relevant compounds. This work will create the first comprehensive genotype-phenotype map for the adrenergic receptor signaling cascade, opening up new avenues for dissecting the molecular basis of gene function and dysfunction in cardiovascular physiology. More broadly, these efforts will provide actionable insights to ultimately guide genotype-based treatment strategies for personalized medicine.
|Program type||Innovative Project Award|
|Effective start/end date||07/01/2019 → 06/30/2021|