title: Aven-mediated checkpoint kinase control regulates proliferation and resistance to chemotherapy in conventional osteosarcoma.
abstract: Conventional high-grade osteosarcoma is the most common primary bone sarcoma, with relatively high incidence in young people. In this study we found that expression of Aven correlates inversely with metastasis-free survival in osteosarcoma patients and is increased in metastases compared to primary tumours. Aven is an adaptor protein that has been implicated in anti-apoptotic signalling and serves as an oncoprotein in acute lymphoblastic leukaemia. In osteosarcoma cells, silencing Aven triggered G2 cell-cycle arrest; Chk1 protein levels were attenuated and ATR-Chk1 DNA damage response signalling in response to chemotherapy was abolished in Aven-depleted osteosarcoma cells, while ATM, Chk2 and p53 activation remained intact. Osteosarcoma is notoriously difficult to treat with standard chemotherapy, and we examined whether pharmacological inhibition of the Aven-controlled ATR-Chk1 response could sensitize osteosarcoma cells to genotoxic compounds. Indeed, pharmacological inhibitors targeting Chk1/Chk2 or those selective for Chk1 synergized with standard chemotherapy in 2D cultures. Likewise, in 3D extracellular matrix-embedded cultures, Chk1 inhibition led to effective sensitization to chemotherapy. Together, these findings implicate Aven in ATR-Chk1 signalling and point towards Chk1 inhibition as a strategy to sensitize human osteosarcomas to chemotherapy.
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, The Netherlands.
authors.forename: Zuzanna
authors.initials: Z
authors.lastname: Baranski
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, The Netherlands.
authors.forename: Tijmen H
authors.initials: TH
authors.lastname: Booij
authors.affiliation: Department of Pathology, Leiden University Medical Centre (LUMC), The Netherlands.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Cleton-Jansen
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, The Netherlands.|OcellO B.V. Leiden, The Netherlands.
authors.forename: Leo S
authors.initials: LS
authors.lastname: Price
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, The Netherlands.
authors.forename: Bob
authors.initials: B
authors.lastname: van de Water
authors.affiliation: Department of Pathology, Leiden University Medical Centre (LUMC), The Netherlands.
authors.forename: Judith V M G
authors.initials: JV
authors.lastname: Bovée
authors.affiliation: Department of Pathology, Leiden University Medical Centre (LUMC), The Netherlands.
authors.forename: Pancras C W
authors.initials: PC
authors.lastname: Hogendoorn
authors.affiliation: Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, The Netherlands.
authors.forename: Erik H J
authors.initials: EH
authors.lastname: Danen
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics* / Q000378:metabolism; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000235:genetics* / Q000378:metabolism; D064007:Ataxia Telangiectasia Mutated Proteins / Q000235:genetics / Q000378:metabolism; D001859:Bone Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D004317:Doxorubicin / Q000494:pharmacology; D059565:G2 Phase Cell Cycle Checkpoints; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012516:Osteosarcoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D015398:Signal Transduction; D013876:Thiophenes / Q000494:pharmacology; D014508:Urea / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: C532363:3-(carbamoylamino)-5-(3-fluorophenyl)-N-(3-piperidyl)thiophene-2-carboxamide; C412506:AVEN protein, human; D048868:Adaptor Proteins, Signal Transducing; D000903:Antibiotics, Antineoplastic; D051017:Apoptosis Regulatory Proteins; D008565:Membrane Proteins; D013876:Thiophenes; D004317:Doxorubicin; D014508:Urea; D011494:Protein Kinases; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1
keywords: 3D cell culture; Aven; Chk1 inhibitors; doxorubicin; osteosarcoma