Abstract 1075: Therapeutic targeting of CBFA2T3-GLIS2 infant AML with ELU001 - folate receptor alpha-directed C’Dot-drug-conjugate
Authors: Le Quy , Chen Feng , Tang Thao , McKay Cynthia Nourigat , Wu Fei , Turker Melik , Gardinier Thomas , Patel Vaibhav , Venkatesan Aranapakam , Khor Tin , Ma Kai , Adams Gregory P. , Meshinchi Soheil 
Published: June 2022

Abstract
Acute myeloid leukemia (AML) in infants is a unique subtype of AML that is driven by oncogenic fusions, resulting in a highly proliferative disease. The CBFA2T3-GLIS2 oncogenic fusion that is seen exclusively in infants and young children &lt;3 years is the most refractory AML with near uniform mortality with conventional therapies. As part of our Target Pediatric AML (TpAML) discovery effort to define novel therapeutic targets, we discovered folate receptor alpha (FRα, FOLR1) transcript to be expressed in CBF-GLIS positive AML and verified cell surface expression of FOLR1 on leukemic cells. We present pre-clinical efficacy of a novel FOLR1 directed therapy (ELU001) in CBF-GLIS AML.ELU001 is a novel ~6 nm-diameter FOLR1-targeted C’Dot-Drug-Conjugate (CDC) incorporating ~21 exatecan molecules as a payload that is designed to treat cancers with improved safety and efficacy as compared to antibody-drug conjugates. A Phase 1 clinical trial of ELU001 in adult patients with tumors that overexpress FOLR1 is currently underway (NCT05001282). In this study, we determined the potential utility of ELU001 for CBFA2T3-GLIS2 AML by evaluating its preclinical efficacy in AML models. We confirmed the binding of ELU001 to AML cells expressing FOLR1 by flow cytometry using MV4;11 AML cell line transduced with an FOLR1 expression construct (MV4;11 FOLR1+). We show that MV4;11 FOLR1+ cells readily bind to ELU001 but not MV4;11 parental cells. Consistent with target-dependent binding, in vitro cytotoxicity studies demonstrated that MV4;11 FOLR1+ cells were highly susceptible to ELU001 (IC50=30 pM) while MV4;11 parental cells were only minimally affected by treatment. To evaluate in vivo efficacy, we transplanted MV4;11 FOLR1+ cells transduced with Luciferase-expression construct into immune deficient NSG mice at 1M cells per mouse and treated the mice the following week with different doses of ELU001 (vehicle, 0.3mg/kg Q3Dx6, 0.5 mg/kg Q3Dx3, and 0.65mg/kg Q3Dx3). Leukemia burden was assessed by whole-body bioluminescence (IVIS) imaging, performed weekly. IVIS imaging showed that ELU001 effectively eradicated the leukemia in vivoat all three doses tested resulting in enhanced survival while mice that received the vehicle treatment exhibited disease progression and eventually succumb to the disease. We confirmed that the leukemia was absent in ELU001-treated mice at the study endpoint of 120 days post treatment by flow cytometric analysis of the bone marrow, spleen and liver tissues harvested at necropsy. ELU001 was well tolerated with an apparent maximum tolerance dose of 1.50 mg/kg/mouse (0.5 mg/kg exatecan, Q3DX3). In summary, we demonstrated that ELU001 is highly effective at eliminating FOLR1 positive AML cells in vitro and in vivo, providing the preclinical data to support further assessment of ELU001 in clinical trials for pediatric patients with CBFA2T3-GLIS2 fusions overexpressing FOLR1.
Citation Format: Quy Le, Feng Chen, Thao Tang, Cynthia Nourigat McKay, Fei Wu, Melik Turker, Thomas Gardinier, Vaibhav Patel, Aranapakam Venkatesan, Tin Khor, Kai Ma, Gregory P. Adams, Soheil Meshinchi. Therapeutic targeting of CBFA2T3-GLIS2 infant AML with ELU001 - folate receptor alpha-directed C’Dot-drug-conjugate [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1075.