Characterization of the novel indolylmaleimides PDA-66 and PDA-377 effect on canine lymphoma cells

// Wen Liu 1, 2, * , Julia Beck 3, * , Laura C. Schmidt 1, 2 , Catrin Roolf 1 , Anahit Pews-Davtyan 4 , Barbara C. Rutgen 5 , Sabine Hammer 6 , Saskia Willenbrock 2 , Anett Sekora 1 , Arndt Rolfs 7, 8 , Matthias Beller 4 , Bertram Brenig 9 , Ingo Nolte 2 , Christian Junghanss 1 , Ekkehard Schutz 3, 9, * , Hugo Murua Escobar 1, 2, * 1 Department of Medicine, Clinic III - Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 2 Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover, Germany 3 Chronix Biomedical, Gottingen, Germany 4 Leibniz-Institute for Catalysis at the University of Rostock, Rostock, Germany 5 Clinical Pathology, Department of Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria 6 Institute of Immunology, Department of Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria 7 Albrecht-Kossel-Institute for Neuroregeneration (Akos), Center for Mental Health, University of Rostock, Rostock, Germany 8 Centogene AG, Rostock, Germany 9 Institute of Veterinary Medicine, Georg-August-University Gottingen, Gottingen, Germany * The authors contributed equally Correspondence to: Hugo Murua Escobar, email: hugo.murua.escobar@med.uni-rostock.de Keywords: PDA, arylindolylmaleimides, canine lymphoma, transcriptome sequencing Received: September 14, 2015 Accepted: April 02, 2016 Published: May 12, 2016 ABSTRACT Protein kinase inhibitors are widely used in chemotherapeutic cancer regimens. Maleimide derivatives such as SB-216763 act as GSK-3 inhibitor targeting cell proliferation, cell death and cell cycle progression. Herein, the two arylindolylmaleimide derivatives PDA-66 and PDA-377 were evaluated as potential chemotherapeutic agents on canine B-cell lymphoma cell lines. Canine lymphoma represents a naturally occurring model closely resembling the human high-grade non-Hodgkin’s lymphoma (NHL). PDA-66 showed more pronounced effects on both cell lines. Application of 2.5μM PDA-66 resulted in a significant induction of apoptosis (approx. 11 %), decrease of the metabolic activity (approx. 95 %), anti-proliferative effect (approx. 85 %) and cell death within 48h. Agent induced mode of action was characterized by whole transcriptome sequencing, 12 h and 24 h post-agent exposure. Key PDA-66-modulated pathways identified were cell cycle, DNA replication and p53 signaling. Expression analyses indicated that the drug acting mechanism is mediated through DNA replication and cycle arrest involving the spindle assembly checkpoint. In conclusion, both PDA derivatives displayed strong anti-proliferation activity in canine B-cell lymphoma cells. The cell and molecular PDA-induced effect characterization and the molecular characterization of the agent acting mechanism provides the basis for further evaluation of a potential drug for canine lymphoma serving as model for human NHL.