2022 MeRIT Primer Participants – Completed Research
Sunday, November 12, 2023
12:45 PM–02:15 PM
Abstract
Introduction: Lung cancer is a leading cause of cancer related deaths. Artesunate (Art) demonstrated anti-cancer activity in established non-small cell lung cancer (NSCLC) cell lines. However, mutations in Kelch-like ECH-associated protein 1 (KEAP1) confers resistance to Art. KEAP1 is known to regulate multiple cellular processes such as oxidative stress response and anti-apoptotic pathways. Furthermore,
KEAP1 loss of function is associated with poor prognosis in NSCLC patients.
Research Question or Hypothesis: Targeting anti-apoptotic pathway sensitives artesunate anti-cancer activity in KEAP1 loss NSCLC.
Study Design: Preclinical evaluation of anticancer activity of Art-drug combination in NSCLC cell lines, A549 (KEAP1 loss) and H1299 (KEAP1 wild type).
Methods: Synergy screening was conducted with several Art-drug combinations. A 6x6 grid method was utilized along with synergyfinder package in R to calculate a ZIP synergy score. CellTiter-Glo 2.0 was used to assess cell viability after 72-hour drug treatment and GraphPadPrism software was used to calculate drug IC50. Protein expressions were assessed with Western blots.
Results: Navitoclax (Nvt), an anti-apoptotic inhibitor, was most synergistic with Art which demonstrated statistically significant mean ZIP synergy scores of 4.37 (p=0.01) and 5.35 (p<0.01) in A549 and H1299, respectively. Mean Art IC50 combined with 5uM Nvt were 4.22uM (95%CI 3.55-5.02) and 1.62uM (95%CI 1.31-2.00) and statistically significantly different (p<0.001 and p<0.01, respectively) compared to single agent Art IC50 of 28.7uM (95%CI 25.2-32.6) and 4.31uM (95%CI 3.24-5.74), in A549 and H1299, respectively. KEAP1 expression decreased in both cell lines treated with Art. NQO1 (KEAP1 downstream marker) expression increased only in H1299 treated with Art. MCL-1 (anti-apoptotic marker) expression increased only in A549 treated with Nvt. BCL-xL (anti-apoptotic marker) expression was unchanged in both cell lines.
Conclusion: Art-Nvt combination is synergistic in NSCLC cell lines, regardless of KEAP1 mutation status. Studies are ongoing to evaluate efficacy of this combination in xenograft murine models and to determine the mechanism of sensitization.
Presenting Author
Keng Hee Peh PharmD, MS, BCOPUniversity of Kentucky
Authors
Jill Kolesar PharmD, MS
University of Kentucky Markey Cancer Center
J. Robert McCorkle PhD
University of Kentucky Markey Cancer Center
Kristen Hill PhD
University of Kentucky Markey Cancer Center