title: The multi-kinase inhibitor TG02 overcomes signalling activation by survival factors to deplete MCL1 and XIAP and induce cell death in primary acute myeloid leukaemia cells.
abstract: The novel multi-kinase inhibitor TG02 has selectivity against cell cycle and transcriptional cyclin dependent kinases (CDKs) as well as fms-like tyrosine kinase receptor-3 (FLT3). Inhibition of transcriptional CDKs preferentially depletes short-lived proteins such as MCL1. We evaluated the in vitro toxicity of TG02 to primary acute myeloid leukaemia (AML) cells in the presence of survival signalling pathway activation by cytokines and fibronectin. One hundred nanomolar TG02 induced a median decrease of 40% in bulk cell survival and 43% in the CD34(+) CD38(-) CD123(+) subset. A 90% inhibitory concentration of 500 nmol/l indicated that TG02 toxicity is not halted by protective cell cycle arrest. Samples with FLT3 internal tandem duplication were not preferentially targeted. By flow cytometry, TG02 treatment caused loss of RNA Polymerase II serine 2 phosphorylation in patient samples, which correlated strongly with BAX activation (R(2) =0·89), suggesting these as potential biomarkers for clinical studies. MCL1 and XIAP expression also decreased. Repeated brief exposure to TG02 in MOLM-13 cells did not result in compensatory up-regulation of survival protein expression. In conclusion, TG02 is potently cytotoxic towards CD34(+) CD38(-) CD123(+)  and bulk AML cells, despite protective signalling pathway activation. This antitumour activity is most likely mediated by dephosphorylation of RNA Polymerase II leading to depletion of survival molecules such as MCL1 and XIAP, with subsequent BAX activation and apoptosis.
authors.affiliation: Academic Haematology, Nottingham University Hospitals City Campus, Clinical Sciences Building, Nottingham, UK. monica.pallis@nottingham.ac.uk
authors.forename: Monica
authors.initials: M
authors.lastname: Pallis
authors.forename: Amina
authors.initials: A
authors.lastname: Abdul-Aziz
authors.forename: Francis
authors.initials: F
authors.lastname: Burrows
authors.forename: Claire
authors.initials: C
authors.lastname: Seedhouse
authors.forename: Martin
authors.initials: M
authors.lastname: Grundy
authors.forename: Nigel
authors.initials: N
authors.lastname: Russell
mesh_terms: D015703:Antigens, CD; D017209:Apoptosis / Q000187:drug effects*; D004354:Drug Screening Assays, Antitumor; D005260:Female; D018922:HL-60 Cells; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology*; D006801:Humans; D015470:Leukemia, Myeloid, Acute; D008297:Male; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D012319:RNA Polymerase II / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D020298:U937 Cells; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C570909:14-methyl-20-oxa-5,7,14,26-tetraazatetracyclo(19.3.1.1(2,6).1(8,12))heptacosa-1(25),2(26),3,5,8(27),9,11,16,21,23-decaene; D015703:Antigens, CD; C494826:BAX protein, human; D006576:Heterocyclic Compounds, 4 or More Rings; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D047428:Protein Kinase Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; D051028:bcl-2-Associated X Protein; D012319:RNA Polymerase II