Analysis of the interplay between all-trans retinoic acid and histone deacetylase inhibitors in leukemic cells.
The treatment of acute promyelocytic leukemia (APL) with all-trans retinoic acid (ATRA) induces granulocytic differentiation. This process renders APL cells resistant to cytotoxic chemotherapies. Epigenetic regulators of the histone deacetylases (HDACs) family, which comprise four classes (I-IV), critically control the development and progression of APL. We set out to clarify the parameters that determine the interaction between ATRA and histone deacetylase inhibitors (HDACi). Our assays included drugs against class I HDACs (MS-275, VPA, and FK228), pan-HDACi (LBH589, SAHA), and the novel HDAC6-selective compound Marbostat-100. We demonstrate that ATRA protects APL cells from cytotoxic effects of SAHA, MS-275, and Marbostat-100. However, LBH589 and FK228, which have a superior substrate-inhibitor dissociation constant (Ki) for the class I deacetylases HDAC1, 2, 3, are resistant against ATRA-dependent cytoprotective effects. We further show that HDACi evoke DNA damage, measured as induction of phosphorylated histone H2AX and by the comet assay. The ability of ATRA to protect APL cells from the induction of p-H2AX by HDACi is a readout for the cytoprotective effects of ATRA. Moreover, ATRA increases the fraction of cells in the G1 phase, together with an accumulation of the cyclin-dependent kinase inhibitor p21 and a reduced expression of thymidylate synthase (TdS). In contrast, the ATRA-dependent activation of the transcription factors STAT1, NF-κB, and C/EBP hardly influences the responses of APL cells to HDACi. We conclude that the affinity of HDACi for class I HDACs determines whether such drugs can kill naïve and maturated APL cells.
Integrated Research and Treatment Center, Center for Sepsis Control and Care (CSCC), Jena University Hospital, Erlanger Allee 101, 07747, Jena, Germany.|Center for Molecular Biomedicine (CMB), Institute of Biochemistry and Biophysics, Friedrich-Schiller-University Jena, Hans-Knöll-Strasse 2, 07745, Jena, Germany.
Katrin
K
Noack
Department of Toxicology, University Medical Center, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany.
Nisintha
N
Mahendrarajah
Cancer and Inflammation Research, Institute for Molecular Medicine, University of Southern Denmark, J.B. Winsløws Vej 25, 5000, Odense C, Denmark.
Dorle
D
Hennig
Ludwig Boltzmann Institute for Cancer Research, Waehringer Strasse 13A, 1090, Vienna, Austria.
Luisa
L
Schmidt
Ludwig Boltzmann Institute for Cancer Research, Waehringer Strasse 13A, 1090, Vienna, Austria.
Florian
F
Grebien
Department of Infectious Diseases, Medical Microbiology and Hygiene, Im Neuenheimer Feld 324, 69120, Heidelberg, Germany.
Dagmar
D
Hildebrand
Department of Toxicology, University Medical Center, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany.
Markus
M
Christmann
Department of Toxicology, University Medical Center, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany.
Bernd
B
Kaina
Institute of Pharmacy, Faculty of Chemistry and Pharmacy, University of Regensburg, 93040, Regensburg, Germany.
Andreas
A
Sellmer
Institute of Pharmacy, Faculty of Chemistry and Pharmacy, University of Regensburg, 93040, Regensburg, Germany.
Siavosh
S
Mahboobi
Department of Infectious Diseases, Medical Microbiology and Hygiene, Im Neuenheimer Feld 324, 69120, Heidelberg, Germany.
Katharina
K
Kubatzky
Center for Molecular Biomedicine (CMB), Institute of Biochemistry and Biophysics, Friedrich-Schiller-University Jena, Hans-Knöll-Strasse 2, 07745, Jena, Germany.
Thorsten
T
Heinzel
Department of Toxicology, University Medical Center, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany. okraemer@uni-mainz.de.
Oliver H
OH
Krämer
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides / Q000494:pharmacology; D022762:CCAAT-Enhancer-Binding Proteins / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004249:DNA Damage / Q000187:drug effects; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D007938:Leukemia / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D016328:NF-kappa B / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D050794:STAT1 Transcription Factor / Q000378:metabolism; D014212:Tretinoin / Q000008:administration & dosage / Q000494:pharmacology*
D001549:Benzamides; D022762:CCAAT-Enhancer-Binding Proteins; D056572:Histone Deacetylase Inhibitors; D016328:NF-kappa B; D011725:Pyridines; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; C118739:entinostat; D014212:Tretinoin
APL; ATRA; DNA damage; HDAC; HDACi; Histone acetylation