Acute myeloid leukemia

Venetoclax and pegcrisantaspase for complex karyotype acute myeloid leukemia

Abstract

Complex karyotype acute myeloid leukemia (CK-AML) has a dismal outcome with current treatments, underscoring the need for new therapies. Here, we report synergistic anti-leukemic activity of the BCL-2 inhibitor venetoclax (Ven) and the asparaginase formulation Pegylated Crisantaspase (PegC) in CK-AML in vitro and in vivo. Ven-PegC combination inhibited growth of multiple AML cell lines and patient-derived primary CK-AML cells in vitro. In vivo, Ven-PegC showed potent reduction of leukemia burden and improved survival, compared with each agent alone, in a primary patient-derived CK-AML xenograft. Superiority of Ven-PegC, compared to single drugs, and, importantly, the clinically utilized Ven-azacitidine combination, was also demonstrated in vivo in CK-AML. We hypothesized that PegC-mediated plasma glutamine depletion inhibits 4EBP1 phosphorylation, decreases the expression of proteins such as MCL-1, whose translation is cap dependent, synergizing with the BCL-2 inhibitor Ven. Ven-PegC treatment decreased cellular MCL-1 protein levels in vitro by enhancing eIF4E-4EBP1 interaction on the cap-binding complex via glutamine depletion. In vivo, Ven-PegC treatment completely depleted plasma glutamine and asparagine and inhibited mRNA translation and cellular protein synthesis. Since this novel mechanistically-rationalized regimen combines two drugs already in use in acute leukemia treatment, we plan a clinical trial of the Ven-PegC combination in relapsed/refractory CK-AML.

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Fig. 1: In vitro anti-AML activity of the combination of Ven and PegC.
Fig. 2: Efficacy of Ven, PegC and Ven-PegC combination in an orthotopic patient-derived xenograft (PDX) model of relapsed AML with complex karyotype.
Fig. 3: Efficacy of single agents Ven, PegC, azacitidine (Aza), and combination of Aza-Ven and Ven-PegC in U937-luc cells.
Fig. 4: Alteration of gene transcription in AML after treatment with Ven, PegC, and Ven-PegC.
Fig. 5: Ven-PegC impedes cap-dependent translation and protein synthesis.
Fig. 6: Ven-PegC impedes cap-dependent translation and protein synthesis.
Fig. 7: Effect of Ven-PegC on plasma amino acid levels in vivo in AML45-luc.
Fig. 8: Pharmacodynamic (PD) effects of Ven-PegC in vivo in AML45-luc.

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Acknowledgements

We thank Ying Zou, MD, PhD, Nicholas Ambulos, PhD and Danielle Sewell, MSRS of the Genomics Shared Service of University of Maryland School of Medicine for validating the cell lines by short tandem repeat analysis and karyotyping them. We thank Xiaoxuan Fan, PhD and Bryan Hahn, BS of the Flow Cytometry Shared Service at UMGCCC for helping to sort the YFP-positive cells after transduction and the Center for Translational Research in Imaging for access to Xenogen. The CRISPR Core at UMGCCC packaged the plasmid into lentiviruses.

Funding

This study was supported by a research grant provided by Jazz Pharmaceuticals to AE. This work was partially supported by the University of Maryland Greenebaum Comprehensive Cancer Center Support grant (P30CA134274) and the State of Maryland’s Cigarette Restitution Funds.

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AE conceived the idea, conceptualized the hypotheses, and designed and supervised all areas of the study. AE, BK, AS, RBG, and RGL designed experiments and developed the methodology. AE, BK, DB, AS, FK, and RGL analyzed and interpreted the data. BK, BB, DB, HK, BC-C, and BSM performed in vitro studies with cell lines and primary cells including transduction of cells. EC, EYC, XM, KMT, and RGL carried out in vivo experiments under IACUC protocols. AM and AS performed the RNA-seq experiments related to transcriptome and translatome. BK performed Western blot and qPCR for mechanistic studies. ETS performed plasma amino acid analysis. FK conducted all the statistical analysis for the in vivo and amino acid analysis studies. AE, BK, and RGL wrote, reviewed, and revised the first draft of the manuscript. SN, MRB, and CIC contributed to preparing and writing the manuscript.

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Correspondence to Ashkan Emadi.

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AE has received research grants from Jazz Pharmaceuticals and NewLink Genetics. AE is a global oncology advisory board member for Amgen and has served as an advisory board member for Genentech and Servier. AE and RGL are Co-Founders and Scientific Advisors for KinaRx, LLC. All other authors declare that they have no relevant competing interests.

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Emadi, A., Kapadia, B., Bollino, D. et al. Venetoclax and pegcrisantaspase for complex karyotype acute myeloid leukemia. Leukemia (2020). https://doi.org/10.1038/s41375-020-01080-6

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