Department of Biological Sciences and the Center for Drug Discovery, Design and Delivery (CD4) at the Southern Methodist University
Opening New Doors in Cancer Research
Amila Nanayakkara, PhD student, Vogel and Wise Lab, Department of Biological Sciences, Southern Methodist University, Dallas, TX
ATP-binding cassette (ABC) transporters move substrates across cell membranes and are of great importance to those researching cancer and cancer therapies. The laboratories of Dr. Pia Vogel and Dr. John Wise in the Department of Biological Sciences and the Center for Drug Discovery, Design and Delivery (CD4) at the Southern Methodist University in Dallas, TX, are particularly interested in the ABC transporters P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP). When expressed in cancer cells, these proteins pump anticancer drugs out of the cells, often rendering chemotherapy of these cancers ineffective. The goal of the Vogel and Wise labs is to discover inhibitors of these pumps that may eventually be used in the clinic to treat cancer patients with therapy-insensitive cancer. The labs employ computational methods to screen for small molecule inhibitors of the ABC transporters and then test the inhibitors in 2D and 3D cell culture models of drug resistant cancers to determine whether the cancer cells can be resensitized to chemotherapeutics.
Amila Nanayakkara is a PhD candidate in the Vogel and Wise labs and has been tasked with conducting and analyzing cell culture studies involved in these screens, including cell viability, migration, apoptosis and drug accumulation/penetration assays. Originally, microplate reader-based assays were used with good success, but during an Experimental Biology annual meeting, Drs. Vogel and Wise were introduced to BioTek’s Cytation™ 5 Cell Imaging Multi-Mode Reader which incorporates both microplate reader functionality and digital widefield microscopy. They were impressed with this dual capability and decided to incorporate one into their research lab.
Once Mr. Nanayakkara became familiar with the instrument and its powerful Gen5™ Microplate Reader and Imager Software, he realized the importance of incorporating both cell population response data using microplate reader-based assays coupled with imaging of the cells. In addition to quantitative data, Mr. Nanayakkara now captures qualitative images to support his findings. Cytation 5 is also used for kinetic studies, taking measurements of an entire 96-well microplate every five minutes for hours or even days. The data is used to determine the rate and mechanism of compound inhibition over time so that slow-acting compounds can be removed from the screens. Simultaneous imaging of all replicas and controls in a plate increases reliability in the comprehensive data.
“Cytation 5 is highly versatile, reliable and really fast.” Mr. Nanayakkara notes. “It makes my work and my graduate studies very easy and efficient.” In fact, the Cytation 5 provided such an influx of information that he gathered data for up to three scientific papers within three months. The first paper, titled, “Targeted inhibitors of P-glycoprotein increase chemotherapeutic-induced mortality of multidrug resistant tumor cells” was published in Scientific Reports (doi: 10.1038/s41598-018-19325-x) and includes significant data collected via Cytation 5.
To learn more about the Center for Drug Discovery, Design, and Delivery visit their web site.
Thanks to Amila Nanayakkara for sharing his BioTek experience.