Abstract 1892: Discovery of targeted therapies for acute myeloid leukemia patients with NUP98-NSD1 and FLT3-ITD
Authors: Kivioja Jarno , Thanasopoulou Angeliki , Kontro Mika , Kumar Ashwini , Yadav Bhagwan , Majumder Muntasir Mamun , Porkka Kimmo , Schwaller Juerg , Heckman Caroline A. 
Published: August 2018

Abstract
NUP98-NSD1 fusion resulting from the t(5;11)(q35;p15.5) is a clinically significant indicator of adverse prognosis in AML. While the understanding of NUP98-NSD1-driven leukaemogenesis has increased over the years, these patients still suffer from highly progressive disease with no effective therapies. To address this challenge, we analyzed primary patient samples and experimental cell models by high-throughput drug screening and RNA sequencing.
Primary samples included bone marrow mononuclear cells (BM MNCs) from three NUP98-NSD1/FLT3-ITD AML patients and ten healthy donors. Experimental cells included lineage-depleted balb/c BM cells transduced with chimeric NUP98-NSD1 and FLT3-ITD retroviruses alone or together, and Ba/F3 cells transduced with human-cloned NUP98-NSD1 lentivirus. The cells were seeded on 384-well plates containing up to 306 approved and investigational drugs in five concentrations over a 10,000-fold range and incubated for 72h at +37°C. Cell viabilities were measured using CellTiter-Glo®. Drug sensitivity scores (DSS) were calculated from area under the dose response curve and select DSS by subtracting DSS of mock-transduced parental cells from the experimental model.
We discovered 14 drugs with highly significant mean DSS difference (p &lt; 0.001) between patients and healthy controls. Further analysis revealed that only two of these drugs (dasatinib and navitoclax) had positive correlation between DSS and BM blast percentage (Pearson's R: 0.83-0.98) as well as higher mean DSS in the context of other AML patients with (n = 9) and without (n = 38) FLT3-ITD. Experimental balb/c cells expressing NUP98-NSD1 had significantly increased sensitivity to BCL2 inhibitors (n = 3) (p &lt; 0.05), while cells co-expressing NUP98-NSD1 and FLT3-ITD had significantly increased sensitivity to FLT3 (n = 11) and MEK inhibitors (n = 6) in comparison to cells expressing FLT3-ITD alone. In pilot combination screen, select efficacy of FLT3 and MEK inhibitors was further increased in the dual positive cells by dasatinib (100 nM). In synergy screens, navitoclax-dasatinib and navitoclax-quizartinib combinations were highly synergistic in primary cells and balb/c cells co-expressing NUP98-NSD1 and FLT3-ITD. Gene expression analysis showed significant up-regulation of dasatinib target genes LCK and FGR, and navitoclax target gene BCL2A1 in the primary AML cells. Interestingly, BCL2A1 was also upregulated in the balb/c BM cells expressing NUP98-NSD1 alone or with FLT3-ITD, but not in cells expressing FLT3-ITD alone.
Taken together, we have identified potential candidate drugs and drug combinations for t(5;11) positive AML. Although the sample size was small, our data suggests that further research into BCL2 inhibitors in combination with TKIs may eventually translate to better survival outlook of AML patients with t(5;11).
Citation Format: Jarno Kivioja, Angeliki Thanasopoulou, Mika Kontro, Ashwini Kumar, Bhagwan Yadav, Muntasir Mamun Majumder, Kimmo Porkka, Juerg Schwaller, Caroline A. Heckman. Discovery of targeted therapies for acute myeloid leukemia patients with NUP98-NSD1 and FLT3-ITD [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1892.