Thermal proteome profiling identifies PIP4K2A and ZADH2 as off‐targets of Polo‐like kinase 1 inhibitor volasertib

• Published in: The FASEB journal 2021-07, Vol.35 (7), p.e21741-n/a
• Main Authors: Goroshchuk, Oksana, Kolosenko, Iryna, Kunold, Elena, Vidarsdottir, Linda, Pirmoradian, Mohammad, Azimi, Alireza, Jafari, Rozbeh, Palm‐Apergi, Caroline
• Format: Article
• Language: English
• Subjects: acute leukemia; Antigens, Surface - metabolism; Cell Cycle Proteins - metabolism; CETSA; Cytarabine - pharmacology; Fatty Acids - metabolism; HL-60 Cells; Humans; Immunity - drug effects; Jurkat Cells; Leukemia, Myeloid, Acute - drug therapy; Leukemia, Myeloid, Acute - metabolism; Medicin och hälsovetenskap; Phosphotransferases (Alcohol Group Acceptor) - metabolism; PIP4K2A; Piperazines - pharmacology; Polo-Like Kinase 1; Protein Kinase Inhibitors - pharmacology; Protein Serine-Threonine Kinases - metabolism; Proteome - metabolism; Proto-Oncogene Proteins - metabolism; Pteridines - pharmacology; Pyrazoles - pharmacology; Quinazolines - pharmacology; thermal proteome profiling; volasertib; ZADH2
• ISSN: 0892-6638; 1530-6860; 1530-6860
• DOI: 10.1096/fj.202100457RR

Polo‐like kinase 1 (PLK1) is an important cell cycle kinase and an attractive target for anticancer treatments. An ATP‐competitive small molecular PLK1 inhibitor, volasertib, has reached phase III in clinical trials in patients with refractory acute myeloid leukemia as a combination treatment with cytarabine. However, severe side effects limited its use. The origin of the side effects is unclear and might be due to insufficient specificity of the drug. Thus, identifying potential off‐targets to volasertib is important for future clinical trials and for the development of more specific drugs. In this study, we used thermal proteome profiling (TPP) to identify proteome‐wide targets of volasertib. Apart from PLK1 and proteins regulated by PLK1, we identified about 200 potential volasertib off‐targets. Comparison of this result with the mass‐spectrometry analysis of volasertib‐treated cells showed that phosphatidylinositol phosphate and prostaglandin metabolism pathways are affected by volasertib. We confirmed that PIP4K2A and ZADH2—marker proteins for these pathways—are, indeed, stabilized by volasertib. PIP4K2A, however, was not affected by another PLK1 inhibitor onvansertib, suggesting that PIP4K2A is a true off‐target of volasertib. Inhibition of these proteins is known to impact both the immune response and fatty acid metabolism and could explain some of the side effects seen in volasertib‐treated patients.