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Blood. 2019 May 10. pii: blood.2018862383. doi: 10.1182/blood.2018862383. [Epub ahead of print]

Hepatic leukemia factor is a novel leukemic stem cell regulator in DNMT3A, NPM1, and FLT3-ITD triple-mutated AML.

Author information

1
Department of Medicine V, Hematology, Oncology and Rheumatology, University Hospital Heidelberg.
2
EMBL, Germany.
3
Universite Laval, Canada.
4
Laboratory of molecular genetics of stem cells, Institute for Research in Immunology and Cancer, Uni.
5
IRIC / Biochemistry, Université de Montréal, Canada.
6
IRIC, Université de montréal, Canada.
7
Department of Medicine V, Hematology, Oncology and Rheumatology, University Hospital Heidelberg, Germany.
8
Department, Hematology and Oncology, University of Halle, Germany.
9
Division of Cancer Epigenomics, German Cancer Research Center (DKFZ), Germany.
10
Clinical Research Division, Fred Hutchinson Cancer Research Center, United States.
11
Terry Fox Laboratory, British Columbia Cancer Agency, Canada.
12
Regeneration in Hematopoiesis, Immunology of Aging, Leibniz-Institute on Aging - Fritz-Lipmann-Institute (FLI), Germany.
13
Research Unit Apoptosis in Hematopoietic Stem Cells, Helmholtz Zentrum München, Germany.
14
Apoptosis in Hematopoietic Stem Cells, Helmholtz Zentrum München, Germany.
15
The Quebec Leukemia Cell Bank and Division of Hematology-Oncology, Canada.
16
Medicine, Hopital Maisonneuve-Rosemont, Canada.
17
Institute for Research in Immunology and Canc, Université de Montréal, Canada.
18
Université Laval, Canada.
19
Hematology / Oncology, Department of Medicine V, Hematology, Oncology and Rheumatology, University Hospital Heidelberg, Germany pabst0404@gmail.com.

Abstract

FLT3, DNMT3A, and NPM1 are the most frequently mutated genes in cytogenetically normal acute myeloid leukemia (AML), but little is known about how these mutations synergize upon co-occurrence. Here we show that triple-mutated AML is characterized by high leukemia stem cell (LSC) frequency, an aberrant leukemia specific GPR56highCD34low immunophenotype, and synergistic upregulation of Hepatic Leukemia Factor (HLF). Cell sorting based on the LSC marker GPR56 allowed isolation of triple mutated from DNMT3A/NPM1 double-mutated subclones. Moreover, in DNMT3A R882 mutated patients, CpG hypomethylation at the HLF transcription start site correlated with high HLF mRNA expression, which was itself associated with poor survival. Loss of HLF 3 via CRISPR/Cas9 significantly reduced the CD34+GPR56+ LSC compartment of primary human triple-mutated AML cells in serial xenotransplantation assays. HLF knockout cells were more actively cycling when freshly harvested from mice, but rapidly exhausted when re-introduced in culture. RNA-sequencing (RNA-Seq) of primary human triple-mutated AML cells after shRNA mediated HLF knockdown revealed the NOTCH target Hairy And Enhancer Of Split 1 (HES1) and the cyclin-dependent kinase inhibitor CDKN1C/p57 as novel targets of HLF potentially mediating these effects. Overall our data establish HLF as a novel LSC regulator in this genetically defined high-risk AML subgroup.

PMID:
31076446
DOI:
10.1182/blood.2018862383

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