title: Atracurium Besylate and other neuromuscular blocking agents promote astroglial differentiation and deplete glioblastoma stem cells.
abstract: Glioblastoma multiforme (GBM) are the most common primary malignant brain tumor in adults, with a median survival of about one year. This poor prognosis is attributed primarily to therapeutic resistance and tumor recurrence after surgical removal, with the root cause suggested to be found in glioblastoma stem cells (GSCs). Using glial fibrillary acidic protein (GFAP) as a reporter of astrocytic differentiation, we isolated multiple clones from three independent GSC lines which express GFAP in a remarkably stable fashion. We next show that elevated expression of GFAP is associated with reduced clonogenicity in vitro and tumorigenicity in vivo. Utilizing this in vitro cell-based differentiation reporter system we screened chemical libraries and identified the non-depolarizing neuromuscular blocker (NNMB), Atracurium Besylate, as a small molecule which effectively induces astroglial but not neuronal differentiation of GSCs. Functionally, Atracurium Besylate treatment significantly inhibited the clonogenic capacity of several independent patient-derived GSC neurosphere lines, a phenomenon which was largely irreversible. A second NNMB, Vecuronium, also induced GSC astrocytic differentiation while Dimethylphenylpiperazinium (DMPP), a nicotinic acetylcholine receptor (nAChR) agonist, significantly blocked Atracurium Besylate pro-differentiation activity. To investigate the clinical importance of nAChRs in gliomas, we examined clinical outcomes and found that glioma patients with tumors overexpressing CHRNA1 or CHRNA9 (encoding for the AChR-α1 or AChR-α9) exhibit significant shorter overall survival. Finally, we found that ex-vivo pre-treatment of GSCs, expressing CHRNA1 and CHRNA9, with Atracurium Besylate significantly increased the survival of mice xenotransplanted with these cells, therefore suggesting that tumor initiating subpopulations have been reduced.
authors.affiliation: Department of Neurological Surgery, Case Western Reserve University School of Medicine and Case Comprehensive Cancer Center, Cleveland, OH, USA.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Spina
authors.affiliation: Department of Neurological Surgery, Case Western Reserve University School of Medicine and Case Comprehensive Cancer Center, Cleveland, OH, USA.
authors.forename: Dillon M
authors.initials: DM
authors.lastname: Voss
authors.affiliation: Department of Pathology, Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Asnaghi
authors.affiliation: Department of Neurological Surgery, Case Western Reserve University School of Medicine and Case Comprehensive Cancer Center, Cleveland, OH, USA.|Department of Neurological Surgery, University Hospital-Case Medical Center, Case Comprehensive Cancer Center, and Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Sloan
authors.affiliation: Department of Neurological Surgery, Case Western Reserve University School of Medicine and Case Comprehensive Cancer Center, Cleveland, OH, USA.
authors.forename: Eli E
authors.initials: EE
authors.lastname: Bar
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D001279:Atracurium / Q000494:pharmacology*; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D005909:Glioblastoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D009466:Neuromuscular Blocking Agents / Q000494:pharmacology*; D011950:Receptors, Cholinergic / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000072056:Transcription Factor HES-1; D023041:Xenograft Model Antitumor Assays
chemical_list: D001279:Atracurium; D051792:Basic Helix-Loop-Helix Proteins; D005904:Glial Fibrillary Acidic Protein; D018398:Homeodomain Proteins; D009466:Neuromuscular Blocking Agents; D011950:Receptors, Cholinergic; D012097:Repressor Proteins; D000072056:Transcription Factor HES-1; C405693:HEY2 protein, human; C078493:HES1 protein, human
keywords: Atracurium Besylate; astrocytic differentiation; glioma; neurotransmitter signaling; stem cells