Multiplexed single-cell contractility assay to develop therapeutics for asthma and hypertension

SUMMARYABSTRACT Asthmatic bronchoconstriction and hypertensive vasoconstriction are extremely common disease states in which excessive contractile cellular forces directly contribute to the pathophysiologyExisting treatments for these diseaseswhich affectmillion andmillion Americansrespectivelyhave severe side effectsbecome desensitized over prolonged useor lack efficacy altogetherIn particularLABAs used in asthma management carry ablack boxwarningandof hypertensive patients require andgtdrugs to control blood pressureDespite understanding the role of cellular force in these scenariosdrug developers have lacked the drug discovery tools that directly target this critically important phenotypeInsteadmany new drug development efforts continue to focus on known pathwaysClearlythere is a significant clinical unmet need in treating resistant asthma and hypertensionand there are large associatedandgt $Bmarkets worldwideSpecificallythere is need to develop new classes of drugs with molecular mechanisms of action that are orthogonal to existing therapies that promote smooth muscle cell relaxation causing bronchodilation or vasodilationForcyte Biotechnologies is an early stage bio pharmaceutical company incubating at UCLA that is leveraging a microtechnology known as FLECSa high throughput screeningHTSplatform that measures contractility of single cells in awellplate formatto identify and bring to market new compound classes that act on force generating pathways within cellsThis is the first and only reported assay that obtains functional force generation data for single cellsat HTS scalesOur initial programs will focus on treatment resistant asthma and hypertensionbut can extend to other diseases associated with abnormal cellular forceIn this proposalwe seek to implement novel multiplexing strategies to extend FLECS s screening bandwidth fromtocell types simultaneouslySpecificallywe will exploit the single cell nature of our platformand combine cell patterning with cell barcoding to enable discrimination of individual cell types from a large mixed population in each well on our wellplatesWe will assess the robustness and separation of the multiplexed signals and develop protocols to maximize signal to noise and reduce both optical and biological cross talkCompletion of our proposed aims will enrich the data generated in screens and substantially reduce the cost per data pointThese enhancements will lay the foundation for successful phase II screens of a proprietarycompound library to identify potential new therapeutics for asthma and hypertension In the proposed workwe will continue development of the first high throughput single cell contractility screening platformbased on fluorescently labeled elastomeric contractible surfacesFLECSas a means for bolstering the pharmaceutical pipeline for asthma and hypertension therapeuticsFLECS currently integrates withwellplate formats to facilitatehigh throughput screening of compounds that act on cellular contractilityThis proposal aims to implement novel spatial and temporal multiplexing strategies to extend FLECS s screening bandwidth tocell types simultaneously to ienrich data generation iisubstantially reduce cost per data point andreduce time to market for potential therapeuticsBy the conclusion of this Phase Ithe platform will be ready to screen a proprietarycompound library for compounds targeting airway and vascular smooth muscle cell contractility in a follow on Phase II SBIR