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Haematologica. 2018 Nov;103(11):1843-1852. doi: 10.3324/haematol.2018.190231. Epub 2018 Jul 12.

Inhibition of protein disulfide isomerase induces differentiation of acute myeloid leukemia cells.

Author information

1
Department of Immunology, Medical University of Warsaw.
2
Laboratory of Experimental Medicine, Center of New Technologies, University of Warsaw.
3
Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw.
4
Postgraduate School of Molecular Medicine, Medical University of Warsaw.
5
Laboratory of Human Cancer Genetics, Center of New Technologies, University of Warsaw.
6
Laboratory of Functional and Structural Genomics, Center of New Technologies, University of Warsaw.
7
Department of Physical Chemistry, Faculty of Pharmacy, Medical University of Warsaw.
8
Laboratory of Imaging Tissue Structure and Function, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw.
9
Laboratory of Mass Spectrometry, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw.
10
Department of Medical Genetics, Center of Biostructure Research, Medical University of Warsaw.
11
Department of Laboratory Diagnostics, Faculty of Health Sciences, Medical University of Warsaw.
12
Department of Experimental Hematooncology, Medical University of Lublin.
13
Department of Experimental Hematology, Institute of Hematology and Transfusion Medicine, Warsaw.
14
Department of Hematology, Oncology and Internal Diseases, Medical University of Warsaw.
15
Faculty of Mathematics and Information Science, Warsaw University of Technology, Warsaw.
16
Department of Immunology, Medical University of Warsaw d.nowis@cent.uw.edu.pl jakub.golab@wum.edu.pl.
17
Center for Preclinical Research and Technology, Medical University of Warsaw.
18
Genomic Medicine, Medical University of Warsaw, Poland.

Abstract

A cute myeloid leukemia is a malignant disease of immature myeloid cells. Despite significant therapeutic effects of differentiation-inducing agents in some acute myeloid leukemia subtypes, the disease remains incurable in a large fraction of patients. Here we show that SK053, a thioredoxin inhibitor, induces differentiation and cell death of acute myeloid leukemia cells. Considering that thioredoxin knock-down with short hairpin RNA failed to exert antiproliferative effects in one of the acute myeloid leukemia cell lines, we used a biotin affinity probe-labeling approach to identify potential molecular targets for the effects of SK053. Mass spectrometry of proteins precipitated from acute myeloid leukemia cells incubated with biotinylated SK053 used as a bait revealed protein disulfide isomerase as a potential binding partner for the compound. Biochemical, enzymatic and functional assays using fluorescence lifetime imaging confirmed that SK053 binds to and inhibits the activity of protein disulfide isomerase. Protein disulfide isomerase knockdown with short hairpin RNA was associated with inhibition of cell growth, increased CCAAT enhancer-binding protein α levels, and induction of differentiation of HL-60 cells. Molecular dynamics simulation followed by the covalent docking indicated that SK053 binds to the fourth thioredoxin-like domain of protein disulfide isomerase. Differentiation of myeloid precursor cells requires the activity of CCAAT enhancer-binding protein α, the function of which is impaired in acute myeloid leukemia cells through various mechanisms, including translational block by protein disulfide isomerase. SK053 increased the levels of CCAAT enhancer-binding protein α and upregulated mRNA levels for differentiation-associated genes. Finally, SK053 decreased the survival of blasts and increased the percentage of cells expressing the maturation-associated CD11b marker in primary cells isolated from bone marrow or peripheral blood of patients with acute myeloid leukemia. Collectively, these results provide a proof-of-concept that protein disulfide isomerase inhibition has potential as a therapeutic strategy for the treatment of acute myeloid leukemia and for the development of small-molecule inhibitors of protein disulfide isomerase.

PMID:
30002127
DOI:
10.3324/haematol.2018.190231
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