Abstract 4668: Patient AML cells and AML cell lines are highly sensitive to CNDAC, the active form of sapacitabine
Authors: Jagan Sucheta , Paganessi Laura A. , Frank Robin R. , Gezer Sefer , Rizman Amy , Keller Margaret C. , Larson Melissa L. , Venugopal Parameswaran , Gregory Stephanie A. , Christopherson Kent W. 
Published: August 2012

Abstract
Introduction: Achieving improvements in survival and reducing relapse remains a challenge in acute myelogenous leukemia (AML) patients. This study evaluated the in vitro efficacy of the active form of novel agent sapacitabine, 2′-C-Cyano-2′-deoxy-1-α-d-arabino-pentofuranosylcytosine (CNDAC, Cyclacel Ltd, Dundee, UK), as compared to current chemotherapeutic drugs Ara-C (Cytarabine) and Mitoxantrone (Mit, synthetic anthracenedione) using two AML cell lines, HL-60 (promyelocytic) and THP-1 (monocytic), as well as bone marrow (BM) and peripheral blood (PB) cells collected from 5 AML patients. Methods: Cells lines and AML patient cells were treated in vitro with Ara-C (1-100 µM), CNDAC (1-100 µM) or Mit (0.005-0.5 µM) for 4 days. During treatment, HL-60, THP-1, and PB AML cells were cultured in suspension. BM AML cells were co-cultured with M2-10B4 mouse stromal cells. Cell lines were assessed immediately. BM and PB AML cells were assessed after an additional 3, 7, or 31 days of co-culture on M2-10B4 cells. Treated cells were assessed for sensitivity (cell death by trypan blue and apoptosis by 7AAD/Annexin V) as compared to untreated cells and IC50 values (50% of maximum possible effective response) were calculated. Results: In HL-60 cells, the amount of cell death was greater with CNDAC compared to Ara-C at all doses tested (p≤0.05, n=3). In THP-1 cells, CNDAC and Mit, but not Ara-C, induced a significant apoptotic response. At a 10-fold lower seeding density, which correlates to higher proliferation rates, the response of THP-1 cells to Ara-C, CNDAC and Mit reached significance compared to untreated cells (p≤0.05, n=3). However, the IC50 values for the 3 drugs in THP-1 cells reflect the observation that this cell line is in essence resistant to AraC (IC50 = 7.77 µM) but sensitive to CNDAC (IC50 = 0.929 µM) and Mit (IC50 = 0.003 µM). Using PB AML cells, a significant response to 1 µM CNDAC and 0.005 µM Mit but not 1 µM Ara-C was observed, as compared to untreated cells at all days post drug removal (p≤0.05, n=5). A significantly greater apoptotic response to CNDAC was observed compared to Ara-C at low doses (p≤0.05, n=5). At higher doses, all 3 drugs induced significant cell death (p≤0.05, n=5). In BM AML cells, overall survival with 1 µM CNDAC or 0.005 µM Mit, but not 1 µM Ara-C, was significantly less than untreated cells 31 days post-treatment (p≤0.05, n=5). Higher doses induced cell death (p≤0.05, n=5) with all 3 drugs. Conclusion: Low dose CNDAC and Mitoxantrone induce a greater response than low dose Ara-C in patient AML cells and AML cell lines. CNDAC also exhibits a greater activity in cell lines (THP-1) that are less sensitive to Ara-C. The in vitro sensitivity of AML cells to CNDAC supports the ongoing clinical evaluation of sapacitabine in AML patients. Future studies are warranted to assess the potential for combining sapacitabine with Ara-C and/or Mitoxantrone, with an emphasis on cells and patients insensitive to Ara-C treatment.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4668. doi:1538-7445.AM2012-4668