Optopatch high throughput all optical electrophysiology

Project Summary In spite of the prevalence and severity of many neurological disorders the development of new classes of drugs has been sluggish for years This is due largely to the lack of good model systems and tools to rapidly study relevant electrical and synaptic phenotypes We aim to overcome these challenges Recent advances in induced pluripotent stem cell iPSC technology reveal the first prospects for studying human neurons paired with clinical histories using fast in vitro technologies The complex electrophysiological behavior of these cell can be recorded with the Optopatch platform and microscope systems recently developed at Q State With these tools it is possible to simultaneously stimulate blue light and record red light electrical activity from around a hundred neurons with one millisecond temporal resolution single cell spatial resolution and high signal to noise This system can be used to measure single cell excitability and firing patters or to probe synaptic transmission by stimulating a subset of neurons with spatially patterned blue light Moving forward we propose to increase microscope throughput without sacrificing capabilities and rigorously test the platform s performance First the microscope will be upgraded with advanced environmental controls well plate compatibility and a fluid handling robot for compound addition Next data storage and analysis will be securely moved to the cloud to handle Terabyte day data rates Once the microscope is fully functional sensitivity and reproducibility well to well plate to plate and batch to batch will be tested using a library of control compounds Finally as a first application we will search for a robust screenable phenotype for Dravet syndrome Neurons will be prepared from ten healthy and ten Dravet patients to look for differences in firing that transcend variation in the genetic background A drug that ameliorates the disease phenotype in the majority of cell lines is a promising candidate to be broadly effective in the clinic A well validated high throughput electrophysiology platform with confirmed phenotypic readouts in human iPSC disease neurons has the potential to change the drug screening landscape for neurological disorders We hope to open a new path to finding treatments for these horrible diseases Project Narrative Progress on new classes of drugs for treating neurological disorders such as epilepsy and ALS has been sluggish for years because of a lack of good model systems and readout technologies Q State uses its engineered proteins and custom microscopes to simultaneously stimulate and record electrical activity from hundreds of human neurons derived from the stem cells of patients with these diseases allowing for a so called disease in a dish model In this application we propose drastically increasing the throughput of our microscope system developed in our successful Phase I project to rapidly screen compounds for treating these debilitating neurological disorders