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New Biological Frontiers Illuminated by Molecular Sensors and Actuators
Poster Abstracts
5-POS
Board 5
Genetically Encoded Tools to Manipulate and Observe Cellular Dynamics in Cardiac
Disease Modelling and Drug Screening
Yu-Fen Chang
, Frances Book, Mark J. Davies, Matthew J. Daniels.
University of Oxford, Oxford, United Kingdom.
Optogenetic technology allows us to control depolarization of electrically excitable cells using
ion-channels that open in response to light. Genetically Encoded Calcium Indicators (GECIs) can
be utilized to visualize cellular response upon opto-stimulation in real time. Previously, we have
successfully demonstrated the use of channelrhodopsin2 (ChR2), a light sensitive cation channel,
with a spectrally compatible red shifted calcium indicator (R-GECO) in a transient expressing
neuronal model. Here, we presented a viral delivery system for the two component parts (an
improved ChR2 and R-GECO) required to target relevant cardiac substrates such as adult guinea
pig cardiomyocytes human-embryonic-stem-cell derived cardiomyocytes (hES-CM), and patient
derived iPS cardiomyocyte derivatives allowing light stimulation with simultaneous Ca++
imaging. To prove the concept of contactless drug screening in excitable cells, known small
molecules were tested on hES-CMs in multi-well plates. Increased Ca++ influx was observed in
hES-CMs upon blue light stimulation after treatment of BayK8644, the L-Type Ca++ channel
agonist. On the other hand, cytosolic calcium fell upon light activation after treating with
BAPTA, a Ca++ chelator. The contactless nature of this technology enables high-throughput
drug screening to be targeted to rapid and reversible phenomena like Ca++ handling, providing a
new platform for cardiac disease modelling.