Page 16 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

References

1. Kepp, O., Galluzzi, L., Lipinski, M., Uan, J. and Kroemer, G. Cell death assays for drug discovery. Nature Reviews 2011 10:221.
2. Abassi, Y.A., Xi, B, Zhang, W., Ye, P., Kirstein, S.L, Gayloard, M.R., Feinstein, S.C., Wang, X. and Xu, X. Kinetic Cell-Based Morphological Screening:
Predisction of Mechanism of Compound Action and Off-target Effects. Chemistry & Biology 2009 16:712.

Recent IncuCyte Publications

Balvers, et. al., investigated the therapeutic benefit of combination therapy in patient-derived glioma stem-like cells (GSC). The current
standard of care for Glioblastoma Multiforme consists of radiation along with temozolomide (TMZ) chemotherapy. A possible strategy
to increase the efficacy of TMZ is through interference with the DNA damage repair machinery, by poly(ADP-ribose) polymerase protein
inhibition(PARPi). Proliferation assays consisted of longitudinal imaging of cell confluency in the IncuCyte system. Phase contrast images
were acquired at 1-3hr intervals and confluence per well was calculated.
ABT-888 enhances cytotoxic effects of temozolomide independent of MGMT status in serum free cultured glioma cells. Balvers, R. K., Lamfers, M.,
Kloezeman, J. J., Kleijn, A., Berghauser Pont, L., Dirven, C. and Leenstra, S. Journal of Translational Medicine 2015 13:74.

Ong, et. al., evaluated PAK1 dysregulation (copy number gain, mRNA and protein expression) in breast cancer tissues. A novel small
molecule inhibitor, FRAX1036, and RNA interference were used to examine PAK1 loss of function and combination with docetaxel in vitro.
The IncuCyte system was used for caspase 3/7 activation apoptosis assays. Cells were treated with DMSO, FRAX1036, and/or docetaxel.
Caspase 3/7 reagent was added and cells imaged every 2 hours or 4 hours for 36 to 72 hours. Data was analyzed using IncuCyte analysis
software to detect and quantify apoptotic cells.

Small molecule inhibition of group I p21-activated kinases in breast cancer induces apoptosis and potentiates the activity of microtubule
stabilizing agents. Ong, C. C., Gierke, S., Pitt, C., Sagolla M., Cheng, C. K., Zhou, W., Jubb, A. M., Strickland, L., Schmidt, M., Duron, S. G., Campbell, D. A.,
Zheng, W., Dehdashti, S., Shen M., Yang, N., Behnke, M. L., Huang W., McKew, J. C., Chernoff J., Forrest, W.F., Haverty, P. M., Chin S., Rakha, E. A., Green, A.
R., Ellis, I. O., Caldas, C., O’Brien, T., Friedman, L. S., Koeppen, H., Rudolph, J., Hoeflich, K. P. Breast Cancer Research 2015 17:59.

Barsyte-Lovejoy, et. al. provide a protocol for measuring early toxicity/apoptosis response to chemical probes using the IncuCyte platform.

Methods in Enzymology, 2016. Chapter 4: Chemical Biology Approaches for Characterization of Epigenetic Regulators. Barsyte-Lovejoy, D., Szewczyk,
M.M., Prinos, P.

Novotny, et. al. evaluated the ability of reversible HER2 inhibitors to inhibit signaling and proliferation in cancer cell lines driven by HER2–
HER3 heterodimers activated via various oncogenic mvechanisms. The IncuCyte platform was used for real-time cell proliferation assays.
CHL-1 cells were plated in clear-bottom black 96-well plates (Corning; 3904) and allowed to adhere overnight. The following day, media
was changed to fresh media that contained either DMSO or compound. Confluence was measured every 2 h for 96 h using two bright-field
images per well.
Overcoming resistance to HER2 inhibitors through state-specific kinase binding. Novotny, C.J., Pollari, S., Park, J.H., Lemmon, M.A., Shen, W., Shokat,
K.M. Nature Chemical Biology 2016 12:923.

Skolekova, et. al., measured caspase-3/7 activity corresponding to the induction of apoptosis in human mesenchymal stromal cells (MSCs)
cultivated in the presence of cisplatin using the IncuCyte platform. Kinetic activation of caspase-3/7 was monitored and quantified using
the IncuCyte® FLR object counting algorithm. The authors showed that the secretory phenotype and behavior of mesenchymal stromal
cells influenced by cisplatin differed from naïve MSC. MSC were more resistant to cisplatin, which was cytotoxic for tumor cells.

Cisplatin-induced mesenchymal stromal cells-mediated mechanism contributing to decreased antitumor effect in breast cancer cells. Skolekova, S.,
Matuskova. M., Bohac, M., Toro, L., Durinikova, E., Tyciakova, S., Demkova, L., Gursky, J., and Kucerova, L. Cell Communication and Signaling. 2016 14:4.

14