This poster will be presented by James Lee, PhD on Sunday December 9th from 1:30 - 3:00 PM. Poster number P1055.
The long term microscopy images of live cell fluorescent reporters expressed on patient derived cell models provide a wealth of big data to study the kinetic phenotypes and biomolecular events of neurological disease states. To discover the kinetic phenotypes of disease processes embedded in induced motor neurons (iMNs), we developed and optimized a functional reporter Sma-GFP that signals the cell apoptosis of iMNs under degeneration. We confirmed, under apoptotic condition, the Smac-GFP is released from mitochondria into cytosol, allowing the complementation with GFP and subsequent emission of fluorescence.
In a longitudinal iMN imaging study using a Nikon BioStation CT, the iMN patient panels are imaged with the Smac-GFP apoptosis and Hb9::RFP motor neuron lineage reporters once every 48 hours for 22 days. We found that the Hb9::RFP reporter successfully identified and tracked the survival of motor neurons in all iMN lines, and the Smac-GFP apoptosis reporter successfully detected neurodegeneration events. Using a machine learning enabled kinetic image analysis tool, we are able to reliably detect both Hb9::RFP motor neuron lineage and the Smac-GFP apoptosis expression for co-expression analysis in iMN image sequences.
We characterized four different kinetic co-expression event types: (1) “co-expression -> neuron death”, (2) “co-expression -> neuron survival”, (3) “non-coexpression -> neuron death”, and (4)“non-coexpression -> neuron survival”. The difference in event type prevalence rates could lead to different neurodegeneration outcomes. We are developing and optimizing three additional cell functional fluorescent reporters for human patient lines. The additional reporters and co-expression analysis could further elucidate kinetic phenotypic states of neurological disease.
Authors: J.S.Lee, T.Cheng, T.Onishi, C.Huang , H.Sasaki, M.Jones, Y.Shi, M.Hattori, J.Ichida, T.Nagai
T.Onishi, M.Hattori and T.Nagai are a part of Department of Biomolecular Science and Engineering,Osaka University,Osaka,Japan
Y.Shi and J.Ichida are a part of Eli and Edy the Broad Center for Regenerative Medicine and Stem Cell Research,University of Southern California, Los Angeles,CA
Other authors are a part of DRVision Technologies LLC, Bellevue, WA