Fangyong Dong1, Michael Eibach1, Jörg W Bartsch1, Amalia M Dolga1, Uwe Schlomann1, Catharina Conrad1, Susanne Schieber1, Oliver Schilling1, Martin L Biniossek1, Carsten Culmsee1, Herwig Strik1, Garrit Koller1, Barbara Carl1, Christopher Nimsky1. 1. 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.).
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.
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: Kyeung Min Joo; Juyoun Jin; Eunhee Kim; Kang Ho Kim; Yonghyun Kim; Bong Gu Kang; Youn-Jung Kang; Justin D Lathia; Kwang Ho Cheong; Paul H Song; Hyunggee Kim; Ho Jun Seol; Doo-Sik Kong; Jung-Il Lee; Jeremy N Rich; Jeongwu Lee; Do-Hyun Nam Journal: Cancer Res Date: 2012-05-22 Impact factor: 12.701
Authors: Gaspar J Kitange; Brett L Carlson; Mark A Schroeder; Patrick T Grogan; Jeff D Lamont; Paul A Decker; Wenting Wu; C David James; Jann N Sarkaria Journal: Neuro Oncol Date: 2008-10-24 Impact factor: 12.300
Authors: Ana Jimenez-Pascual; James S Hale; Anja Kordowski; Jamie Pugh; Daniel J Silver; Defne Bayik; Gustavo Roversi; Tyler J Alban; Shilpa Rao; Rui Chen; Thomas M McIntyre; Giorgio Colombo; Giulia Taraboletti; Karl O Holmberg; Karin Forsberg-Nilsson; Justin D Lathia; Florian A Siebzehnrubl Journal: Cancer Discov Date: 2019-08-21 Impact factor: 39.397