title: The metalloprotease-disintegrin ADAM8 contributes to temozolomide chemoresistance and enhanced invasiveness of human glioblastoma cells.
abstract: BACKGROUND
  Despite multimodal treatment, glioblastoma (GBM) therapy with temozolomide (TMZ) remains inefficient due to chemoresistance. Matrix metalloproteinase (MMP) and a disintegrin and metalloprotease (ADAM), increased in GBM, could contribute to chemoresistance and TMZ-induced recurrence of glioblastoma.
  
  
  METHODS
  TMZ inducibility of metalloproteases was determined in GBM cell lines, primary GBM cells, and tissues from GBM and recurrent GBM. TMZ sensitivity and invasiveness of GBM cells were assessed in the presence of the metalloprotease inhibitors batimastat (BB-94) and marimastat (BB-2516). Metalloprotease-dependent effects of TMZ on mitochondria and pAkt/phosphatidylinositol-3 kinase (PI3K) and phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) pathways were analyzed by fluorescence activated cell sorting, morphometry, and immunoblotting. Invasiveness of GBM cells was determined by Matrigel invasion assays. Potential metalloprotease substrates were identified by proteomics and tested for invasion using blocking antibodies.
  
  
  RESULTS
  TMZ induces expression of MMP-1, -9, -14, and ADAM8 in GBM cells and in recurrent GBM tissues. BB-94, but not BB-2516 (ADAM8-sparing) increased TMZ sensitivity of TMZ-resistant and -nonresistant GBM cells with different O(6)-methylguanine-DNA methyltransferase states, suggesting that ADAM8 mediates chemoresistance, which was confirmed by ADAM8 knockdown, ADAM8 overexpression, or pharmacological inhibition of ADAM8. Levels of pAkt and pERK1/2 were increased in GBM cells and correlated with ADAM8 expression, cell survival, and invasiveness. Soluble hepatocyte growth factor (HGF) R/c-met and CD44 were identified as metalloprotease substrates in TMZ-treated GBM cells. Blocking of HGF R/c-met prevented TMZ-induced invasiveness.
  
  
  CONCLUSIONS
  ADAM8 causes TMZ resistance in GBM cells by enhancing pAkt/PI3K, pERK1/2, and cleavage of CD44 and HGF R/c-met. Specific ADAM8 inhibition can optimize TMZ chemotherapy of GBM in order to prevent formation of recurrent GBM in patients.
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Fangyong
authors.initials: F
authors.lastname: Dong
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Michael
authors.initials: M
authors.lastname: Eibach
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Jörg W
authors.initials: JW
authors.lastname: Bartsch
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Amalia M
authors.initials: AM
authors.lastname: Dolga
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Uwe
authors.initials: U
authors.lastname: Schlomann
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Catharina
authors.initials: C
authors.lastname: Conrad
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Susanne
authors.initials: S
authors.lastname: Schieber
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Oliver
authors.initials: O
authors.lastname: Schilling
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Martin L
authors.initials: ML
authors.lastname: Biniossek
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Carsten
authors.initials: C
authors.lastname: Culmsee
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Herwig
authors.initials: H
authors.lastname: Strik
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Garrit
authors.initials: G
authors.lastname: Koller
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Barbara
authors.initials: B
authors.lastname: Carl
authors.affiliation: Department of Neurosurgery, Philipps-University Marburg, Marburg, Germany (F.D., M.E., J.W.B., U.S., C.Co., S.S., B.C., C.N.); Department of Neurosurgery, Tongji Hospital, Wuhan, China (F.D.); Philipps-University Marburg, Institute for Pharmacology and Clinical Pharmacy, Marburg, Germany (A.M.D., C.Cu.); Institute of Molecular Medicine and Cell Research, University of Freiburg, Freiburg, Germany (O.S., M.L.B.); BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany (O.S.); Department of Neurology, Philipps-University Marburg, Marburg, Germany (H.S.); Biomaterials, Biomimetics and Biophotonics Research Group, King's College London Dental Institute, London, United Kingdom (G.K.).
authors.forename: Christopher
authors.initials: C
authors.lastname: Nimsky
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000201:enzymology / Q000473:pathology*; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D003606:Dacarbazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D031541:Fluorescence Resonance Energy Transfer; D005909:Glioblastoma / Q000201:enzymology / Q000473:pathology*; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins / Q000378:metabolism*; D009361:Neoplasm Invasiveness / Q000473:pathology; D060888:Real-Time Polymerase Chain Reaction; D000077204:Temozolomide; D059467:Transcriptome / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D008565:Membrane Proteins; D003606:Dacarbazine; D051722:ADAM Proteins; C068490:ADAM8 protein, human; D000077204:Temozolomide
keywords: TMZ; chemoresistance; glioblastoma; hydroxamate inhibitors; metalloproteases
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