Abstract 6038: Combined targeting of E-selectin/CXCR4 and FLT3 by GMI-1359 and sorafenib effectively reduces leukemia cell burden and protects normal hematopoiesis in a patient-derived AML xenograft model
Authors: Zhang Weiguo , Chang Kyung Hee , Basyal Mahesh , Jia Yannan , Ostermann Lauren , Fogler William E. , Magnani John L. , Andreeff Michael 
Published: September 2020

Acute myelogenous leukemia (AML) is characterized by an accumulation of abnormal white blood cells. Internal tandem duplications in the Fms-like tyrosine kinase 3 (FLT3-ITD) account for 30% of adult AML cases and confer poor prognosis (Kottaridis et al., 2003; Thiede et al., 2002). FLT3 inhibitors like sorafenib efficiently eliminate leukemia blast in the peripheral blood (PB), but frequently not in the bone marrow (BM) (Zhang et al., 2008). This suggests a protective effect of the BM on leukemic stem cells, which is mediated by E-selectin and CXCL12 expression in endothelial cells (ECs) and mesenchymal stem cells (MSCs) in the BM vascular and endosteal niches (Horacek et al., 2013; Peled and Tavor, 2013). Our previous study demonstrated that targeting E-selectin/CXCR4 with the dual E-selectin/CXCR4 antagonist GMI-1359 markedly reduced leukemia cell adhesion to ECs and MSCs and eliminated the BM-mediated protection of leukemic cells during FLT3-targeted therapy in vitro, and effectively reduced leukemia cell in the BM in vivo (Zhang et al., 2016). Further, GMI-1359 combined with cytarabine/daunorubicin provided a profound survival benefit in FLT3-mutated leukemia cell MV4-11-bearing mice (Zhang et al., 2015). In the preset study, we compared expression levels of E-selectin ligands and CXCR4 in FLT3 inhibitor-sensitive Ba/F3-FLT3-ITD cells and their inhibitor-resistant counterparts Ba/F3-FLT3-ITD+D835Y and Ba/F3-FLT3-ITD+F691L. The resistant cells expressed 1.7 to ~5.6-fold higher E-selectin ligand and 10-fold higher CXCR4 levels. In addition, BM-mimetic hypoxia culture profoundly upregulated P-selectin glycoprotein ligand-1 (PSGL-1) and CXCR4 levels on leukemia cells. PSGL-1 is also a high-efficiency ligand for E-selectin. We then evaluated anti-leukemia effects of co-targeting E-selectin/CXCR4 and FLT3 with GMI-1359 and sorafenib in a patient-derived AML xenograft model (harboring FLT3-ITD and WT1 mutations). Leukemia cell engraftment was evaluated using FACS and immunohistochemistry by hCD45 staining. We observed that GMI-1359 efficiently mobilized leukemic cells into PB and in combination with sorafenib enhanced the anti-leukemia activity and significantly reduced leukemia cell infiltration by 92%, 82%, 69% and 45% in liver, lung, spleen and BM, respectively, as compared with vehicle-treated mice (p &lt; 0.05). The GMI-1359/sorafenib combination improved mouse survival (median survival were 109, 87, 126 and 138.5 days for the vehicle, GMI-1359, sorafenib and combination groups, respectively). Importantly, we observed that the combination protected normal hematopoiesis by increasing the number of BM megakaryocytes, myelocytes, and erythrocytes. The underlying mechanism(s) for this effect is under investigation. Our findings suggest that co-targeting E-selectin/CXCR4/FLT3 exerts remarkable protection of normal hematopoiesis in addition to more efficiently reducing leukemia cells.

Citation Format: Weiguo Zhang, Kyung Hee Chang, Mahesh Basyal, Yannan Jia, Lauren Ostermann, William E. Fogler, John L. Magnani, Michael Andreeff. Combined targeting of E-selectin/CXCR4 and FLT3 by GMI-1359 and sorafenib effectively reduces leukemia cell burden and protects normal hematopoiesis in a patient-derived AML xenograft model [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6038.