A UVA Wise senior and a sophomore spent the summer researching whether ruthenium organometallic compounds are a viable alternative to platinum-based chemotherapeutic agents. In other words, Olivia Fast, the senior, and Brittany Gentry, the sophomore, decided to see if the ruthenium compounds would cause DNA damage in human cells. The results were interesting, to say the least. So interesting, in fact, that the students, along with Mike Shell, Assistant Professor of Biology, and Floyd Beckford, Van Daniel III Endowed Chair of Chemistry, had their research published in the journal Bioscience Reports.
“Going into the project, I had no idea what to expect,” Gentry said.
Gentry had just graduated high school and was part of the Upward Bound Bridge program, a program that gives students an extra boost to begin their college career. She was selected to do an internship with Professor Shell because he often works with Upward Bound Bridge students.
One of the first things Gentry was tasked with in the lab was to work with Fast on immunofluorescence microscopes to take photographs of cells. Using fluorescently-labeled antibodies, Gentry was able to identify specific sites where DNA is damaged in human cells. When she saw little green dots in the photos, she knew that it revealed DNA cell damage. The slides were set up as a blind test, so Gentry did not have any additional information on what the slides represented.
“I was surprised about how much I learned in a short time,” Gentry said. “We were looking to see if these compounds induced damage in cells. I was using new equipment, and I had never written a manuscript. I learned a lot more than I expected. It was just two weeks after I graduated high school and I was already doing research.”
Professor Shell, she said, taught her how to take a problem and apply it to the real world. It wasn’t long before he had her training others on the use of the microscope because she got good at that particular task.
Fast said the research path always seemed intimidating, but she thought of her future plans during her sophomore year and decided to ask Professor Shell how to get involved in research. She wrote a proposal for a Fellowship in Natural Sciences and it was accepted. She admits it was still intimidating when she got started because there was a lot of laboratory techniques she needed to learn. Fast optimized the MTT metabolism assay to test for cytotoxic effects of exposure to the test compounds.
“I had to be able to do it on my own, and I made mistakes,” Fast said. “I killed all of the cells once. I had to learn as I went.”
Today, Fast has been published twice for her research. Her work on this project has also led to a new collaboration between the Shell laboratory and researchers at Tusculum University. She plans to enter dental school once she graduates.
“I’ve had opportunities here that I would not have had elsewhere,” she said. “The research I’ve done will certainly help my future. It is out of my comfort zone, but doing research is different than taking a lab class. You get some guidance but you get to make decisions on your own, which means you grow as a scientist. My confidence level grew in the lab.”
After plenty of work in the laboratories of the Sandridge Science Center, the biochemistry majors reported the cells were, indeed, damaged by exposure to polynuclear ruthenium organometallic compounds. Additionally, the DNA damage was more cytotoxicin human cells that were completely deficient in nucleotide excision repair. In other words, Fast and Gentry noted that the results of their research suggests that the compounds do induce DNA damage in cells, and that cell resistance to the compounds may be influenced by the NER phenotype of the cells.
At the end of the project, Gentry and Fast concluded that up-regulation of NER activity can lead to cancer cells becoming resistant to chemotherapeutics. Which is why it is important to understand the role of DNA repair in cytotoxic mechanisms in improving compound efficacy.
“This is the first evidence that exposure to these ruthenium compounds results in DNA damage in cells,” the students concluded in their published report, though the exact type of DNA damage could not be classified. This is an important finding because these compounds, synthesized and characterized in the Beckford laboratory by former UVA Wise student Madison Niece, bind to but do not form covalent bonds with isolated DNA. It is possible, they concluded, that the compounds actually undergo metabolic activation within the cell or possibly stimulate the production of secondary toxicants.
“The overall point was to visualize areas of DNA damage,” Fast said. “Our results showed that exposure to the compounds resulted in areas of DNA damage. The research we did is a starting point for chemotherapeutic compounds. You have to know what they are going to do to the cell before you can do anything else. This is the starting point, and that is really important.”
To read more about their research, visit http://www.bioscirep.org/content/ppbioscirep/39/7/BSR20190378.full.pdf
Photo of Olivia Fast