High-throughput screening of vasoprotectors in the brain using single-cell force mapping towards anti-migraine therapies

SUMMARYABSTRACT Migraine headaches affect up toof people world wideincluding twice as many women as menand have been called the single most debilitating neurological disorder in the worldExisting firstand second line treatments including NSAIDS and triptans haveat bestmixed results in the general population as more than half of respondents from a study of andgtmigraine patients reported unsatisfactory pain management with existing treatment optionsClearlythere is a significant clinical unmet need in treating migraine more effectively and a large correspondingandgt $ Bmarket worldwideSpecificallythere is need to develop new classes of drugs with molecular mechanisms of action that are orthogonal to existing therapies that reverse or prevent vasodilation of intracranial vascular smooth muscle cellsForcyte Biotechnologies is an early stage bio pharmaceutical company in Los Angeles that has partnered with advanced high throughput screening and laboratory automation as well as nanofabrication facilities at UCLAthat is leveraging a microtechnology known as FLECSa highthroughput 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 existing programs have focused on treatment resistant asthma and hypertensionand have already had success in discovering novel phenotypic modulators relating to these indications In this proposalwe will build off of our innovative single cell phenotypic screening platform to develop a functional assay for screening and developing vasoprotective compounds to treat migraineOur approach replaces older intact artery migraine modelswhich are extremely informative but very low throughputwith a miniaturized and automated cell contractility assay implemented in thewellplate formatThis will increase throughput from only a few experiments to up toexperiments per day! Such a transformative improvement will enable us to screen entire compound librariesa feat previously impossibleand greatly improve the probability of successful discoveryFurthermorethe proposed assay would offer high throughput functional follow up to other target based affinity assays that cannot predict phenotypic activityOnce developedwe will focus our assay on high throughput screens of unique natural product librariesa rich but relatively underexplored compound space due to throughput limitations of existing methods for evaluating vascular tone and responsivenessA follow on Phase II proposal will perform screen an expanded natural product library and perform selectivity counter screens in related cell types to establish a deep safety profile for the discovered hitsto facilitate transitions to later stages of drug development NARRATIVE In the proposed workwe will build off our platform technologythe first high throughput single cell contractility screening platform based on fluorescently labeled elastomeric contractible surfacesFLECSto develop a functional assay for screening and developing vasoprotective compounds to treat migrainewhich is one of the most debilitating and prevalent disorders todayOur approach replaces older intact artery migraine modelswhich are extremely informative but very low throughputwith a miniaturized and automated cell contractility assay implemented in thewellplate formatThis will increase throughput from a few experiments to up toexperiments per daySuch a transformative improvement will enable us to screen entire compound librariesa feat previously impossibleand greatly improving the probability of successful discoveryFurthermorethe proposed assay would offer high throughput functional follow up to other target based affinity assays that cannot predict phenotypic activityUpon completionwe will focus screening efforts on the rich but under explored natural product spaceto elucidate new therapeutic mechanisms for migraine towards development of novel therapies