Abstract 2731: Impact of hypomethylating agents on hTERT expression and synergistic effect in combination with imetelstat, a telomerase inhibitor, in AML cell lines

Abstract

Abstract The catalytic subunit of human telomerase (hTERT) is highly expressed in acute myeloid leukemia (AML). Transcriptional factors and epigenetic modifications regulate this expression. Studies suggest hypermethylation promotes hTERT overexpression. Imetelstat is a competitive telomerase inhibitor. Decitabine (DAC) and azacitidine (AZA) are DNA methyltransferase inhibitors (DNMTIs) used in the treatment of AML. Combining imetelstat and DNMTI may be an improved treatment option for AML. We investigated effects of DAC and AZA on hTERT expression and cell viability in AML cell lines OCI-AML3 and OCI-AML5, and evaluated growth inhibition when combined with imetelstat. OCI-AML3 had higher baseline expression of hTERT compared with OCI-AML5. Dose-dependent downregulation of hTERT expression was observed with DAC and AZA treatments. OCI-AML5 was more sensitive to DAC (1 μM, 46% viability) than OCI-AML3 (1 μM, 95% viability) whereas sensitivity to AZA was similar in both cell lines (1 μM, 29% vs 33% viability). After removal of the drugs, growth inhibition was not sustained and cell proliferation rebounded. Single-agent imetelstat reduced both hTERT expression and telomerase activity but resulted in only limited growth inhibition in these cell lines. It was hypothesized that combining DNMTI with imetelstat may exert more potent growth inhibition. To test this, both cell lines were treated with DAC or AZA for 72 hours followed by different concentrations of imetelstat for 2 to 4 weeks. Dose-dependent synergistic activity was demonstrated for DAC followed by imetelstat: OCI-AML3 cell viability changed from 95% with DAC to 61% and 10% after 2 and 4 weeks of imetelstat, respectively; OCI-AML5 cell viability changed from 46% with DAC to 6% and 1% after 2 and 4 weeks of imetelstat, respectively. In contrast, no enhanced activity was observed in either cell line with AZA followed by imetelstat, but imetelstat was able to slow the growth rebound after AZA removal. Additional studies are ongoing to further understand the mechanism of action of this drug combination in AML and to evaluate different combination sequences of DNMTI with imetelstat for optimal anticancer activity. In summary, DAC and AZA reduced hTERT expression and differentially inhibited cell viability in AML cell lines. Although at the same concentration AZA was more active than DAC in OCI-AML3 and OCI-AML5 lines, the sustainability of growth inhibition after washout differed. Growth rebounded after removal of AZA or DAC treatment. Dose-dependent synergistic activity demonstrated by DAC followed by imetelstat was not seen with AZA followed by imetelstat. Distinctly different mechanisms of action of DAC and AZA may underlie their differential activity when combined with imetelstat, warranting further investigation. Citation Format: Joshua Rusbuldt, Jacqueline Bussolari, Aleksandra Rizo, Fei Huang. Impact of hypomethylating agents on hTERT expression and synergistic effect in combination with imetelstat, a telomerase inhibitor, in AML cell lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2731.