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Inhibition of DOT1L and PRMT5 promote synergistic anti-tumor activity in a human MLL leukemia model induced by CRISPR/Cas9

Abstract

MLL rearrangements play a crucial role in leukemogenesis and comprise a poor prognosis. Therefore, new treatment strategies are urgently needed. We used the CRISPR/Cas9 system to generate an innovative leukemia model based on 100% pure MLL-AF4 or -AF9 rearranged cells derived from umbilical cord blood with indefinite growth in cell culture systems. Our model shared phenotypical, morphological and molecular features of patient cells faithfully mimicking the nature of the disease. Thus, it serves as a fundamental basis for pharmacological studies: inhibition of histone methyltransferase disruptor of telomeric silencing 1-like (DOT1L) is one specific therapeutic approach currently tested in clinical trials. However, success was limited by restricted response warranting further investigation of drug combinations. Recently, it has been shown that the inhibition of protein arginine methyltransferase 5 (PRMT5) exhibits anti-tumoral activity against human cell lines and in MLL mouse models. Here, we used DOT1L and PRMT5 inhibitors in our human MLL-rearranged model demonstrating dose-dependent reduced proliferation, impairment of cell cycle, increasing differentiation, apoptosis, downregulation of target genes and sensitization to chemotherapy. Strikingly, the combination of both compounds led to synergistic anti-tumoral effects. Our study provides a strong rationale for novel targeted combination therapies to improve the outcome of MLL-rearranged leukemias.

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Acknowledgements

We thank the Flow Cytometry Core Facility Berg of the University Hospital Tuebingen for their excellent technical support. Likewise, we would like to thank the Department of Obstetrics and Gynecology of the University Hospital Tuebingen for providing human cord blood, and the core facility c.ATG of the University Hospital Tuebingen for preparing the RNA-seq libraries, sequencing the samples and data quality control. Furthermore, we thank Dr. Johan Jeong for his technical support in the analysis of the RNA-seq data. CS was supported by a Junior Research Group Grant of the Interdisciplinary Centre for Clinical Research (IZKF, 2383-0-0), the Wuerttembergischer Krebspreis and the Clinician Scientist Program of the Faculty of Medicine Tuebingen. DS was supported by a Max Eder Junior Research Group Grant from the Deutsche Krebshilfe, a New Investigator Award of the American Society for Blood and Marrow Transplantation (ASBMT), a Junior Research Group Grant of the Interdisciplinary Centre for Clinical Research (IZKF, 2316-0-0) and the Clinician Scientist Program of the Faculty of Medicine Tuebingen. JMSH was supported by a Margarete-von-Wrangell fellowship through the Ministry of Science, Research and the Arts Baden-Wuerttemberg, a Junior Research Group Grant of the Interdisciplinary Centre for Clinical Research (IZKF, 2386-0-0) and, together with TH, received funding from the decipherPD transnational consortium on Epigenomics of Complex Diseases (BMBF grant number 01KU1503).

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KAS and CS designed and performed the research, analyzed data and wrote the paper. HK, SDS, HS, DS, TH, JMSH, BM, and FF performed research and analyzed data. All authors edited the paper for content.

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Correspondence to Corina Schneidawind.

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Secker, KA., Keppeler, H., Duerr-Stoerzer, S. et al. Inhibition of DOT1L and PRMT5 promote synergistic anti-tumor activity in a human MLL leukemia model induced by CRISPR/Cas9. Oncogene 38, 7181–7195 (2019). https://doi.org/10.1038/s41388-019-0937-9

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