Developing an in vitro model for cancer pain using all-optical electrophysiology

Project Summary Cancer pain therapeutics comprise more than one quarter of the total pain marketThe current treatment paradigm relies on opioidswhich are not effective for many patients and can have debilitating side effects for othersDespite the clear societal need for better pain medicinesthere has been glacial progress in bringing new drugs to marketThis shortfall is due in part to the lack of translatable model systems and tools to rapidly study relevant electrical and synaptic phenotypes associated with painWe aim to overcome these challengesThe all optical electrophysiology platform Optopatchrecently developed at Q State Biosciences and comprised of engineered optogenetic proteinscustom microscopesand softwaremakes it possible to simultaneously stimulateblue lightand recordred lightelectrical activity from around one hundred neurons with one millisecond temporal resolutionsingle cell spatial resolution and high signal to noiseThe Optopatch platform can be used to study single cell excitabilitysynaptic transmissionand network behavior with information content comparable to manual patch clampbut at dramatically higher throughputIn this applicationwe propose to create an in vitro model of cancer pain via chemotherapeutics or mimicking the local tumor chemical environment experienced by sensory neuronsWe will establish an Optopatch synaptic assay between primary rodent DRG neurons and dorsal horn neurons that will serve as the basis for phenotypic evaluationUsing our Optopatch excitability and synaptic assayswe will develop acancer pain soupthat consists of the relevant signaling moleculesThis formulation will be validated in vitro using human stem cell derived sensory neurons as well as in vivo with nociception assays in rodentsWe hypothesize that addition of this chemical mixture will lead to a phenotype of hyperexcitability or increased synaptic transmissionWe will screen established tool compounds and attempt to reverse the phenotypeThe Phase I application will establish the cancer pain in vitro response that can be used for phenotypic drug screening efforts in Phase II to identify novel therapeutics for cancer pain Project Narrative Effective treatment of cancer pain remains a large unmet medical needas the current standard of care relies on opioids that lack efficacy and lead to debilitating side effectsQ State has created a platform with engineered optogenetic proteins and custom microscopes to simultaneously stimulate and record electrical activity from hundreds of human or rodent sensory neuronsallowing for a so calleddisease in a dishmodel of painIn this applicationwe propose to create an in vitro model of cancer pain by testing for a hyperexcitability phenotype upon treatment of sensory neurons with either chemotherapeutics or a cocktail of signaling molecules that mimic the tumor local chemical environmentthese models will enable future drug screening efforts to identify novel therapeutics for cancer pain