Hello! My name is Daniel Grammer, and I am an AP Research student at Rock Canyon High School in Highlands Ranch, Colorado. I will be investigating how the addition of the antibiotic Novobiocin Sodium Salt to a PARP inhibitor treatment of RIN-m Rat Beta Cells with Insulinoma impacts the health of the cell culture. PARP inhibitors causing fatal oxidative stress in a subset of tumor cells with deficiencies in DNA repair. PARP inhibitor resistance in tumor cells is an emerging problem, and the inhibition of the DNA repair by Novobiocin Sodium Salt is a potential pathway for reversing resistance. This research will contribute to the academic conversation around increasing PARP inhibitor efficacy in tumorigenic cells, in order to improve their cancer treatment potential.

Abstract

Targeted cancer therapies are a class of cancer therapies that target genetic deficiencies in cancer cells. One such therapy, PARP inhibitors, target homologous recombination repair (HRR) deficient cancer by preventing the formation of the PARP1 complex, preventing HRR and inducing apoptosis. Novel research suggests that coumarin compounds, such as Novobiocin sodium salt, might enhance the efficacy of PARP inhibitors through the inhibition of a secondary pathway to HRR. This study will examine the efficacy of Olaparib (PARP inhibitor), Novobiocin, and dual treatment on cell viability in RIN-m islet cells with insulinoma (a benign pancreatic tumor). It is hypothesized that the dual treatment will decrease total cell viability the most in comparison to a no-treatment negative control, because Novobiocin inhibits DNA polymerase theta, enhancing total HRR inhibition. A 10 mM solution of Olaparib and a 300 uM solution of Novobiocin will each be tested alone and in combination in 24 hour exposure trials. A Trypan blue exclusion assay will be used to determine the number of viable cells in each trial. The three treatment groups will be compared to a no-treatment negative control of RIN-m cells. NIH/3T3 cells will be used to verify targeted tumorigenic toxicity. Three trials will be run for each group, and a two-proportion z-test will be used to determine statistical significance between the treatment and control groups. Biologics, including the RIN-m and NIH/3T3 cell lines, are approved for the Rock Canyon High School (RCHS) BSL-1 laboratory. All chemicals have been approved by RCHS’s Chemical Safety Manager Kerry Hinton.