title: Extracellular vesicles derived from human vestibular schwannomas associated with poor hearing damage cochlear cells.
abstract: Vestibular schwannoma (VS) is a tumor of the vestibular nerve that transmits balance information from the inner ear to the brain. Sensorineural hearing loss occurs in 95% of patients with these tumors, but the cause of this loss is not well understood. We posit a role of VS-secreted extracellular vesicles (EVs) as a major contributing factor in cochlear nerve damage.
  
  
  
  Using differential centrifugation, we isolated EVs from VS cell line HEI-193 and primary cultured human VS cells from patients with good hearing or poor hearing. The EVs were characterized using a Nanosight device and transmission electron microscopy and by extracting their RNA content. The EVs' effects on cultured murine spiral ganglion cells and organotypic cochlear cultures were studied using a transwell dual-culture system and by direct labeling of EVs with PKH-67 dye. EV-induced changes in cochlear cells were quantified using confocal immunohistochemistry. Transfection of VS cells with a green fluorescent protein-containing plasmid was confirmed with reverse transcription PCR.
  
  
  
  Human VS cells, from patients with poor hearing, produced EVs that could damage both cultured murine cochlear sensory cells and neurons. In contrast, EVs derived from VS cells from patients with good hearing did not damage the cultured cochlear cells.
  
  
  
  This is the first report on EVs derived from VSs and on the capacity of EVs from VSs from patients with hearing loss to selectively damage cochlear cells, thereby identifying a potential novel mechanism of VS-associated sensorineural hearing loss.
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Vitor Y R
authors.initials: VY
authors.lastname: Soares
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Nadia A
authors.initials: NA
authors.lastname: Atai
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Takeshi
authors.initials: T
authors.lastname: Fujita
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Sonam
authors.initials: S
authors.lastname: Dilwali
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Sarada
authors.initials: S
authors.lastname: Sivaraman
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Lukas D
authors.initials: LD
authors.lastname: Landegger
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Fred H
authors.initials: FH
authors.lastname: Hochberg
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Carlos A P C
authors.initials: CA
authors.lastname: Oliveira
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Fayez
authors.initials: F
authors.lastname: Bahmad
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Xandra O
authors.initials: XO
authors.lastname: Breakefield
authors.affiliation: Department of Otolaryngology, Eaton Peabody Laboratories, Massachusetts Eye and Ear, Boston, Massachusetts (V.Y.R.S., T.F., S.D., L.D.L., K.M.S.); Department of Otology and Laryngology, Harvard Medical School, Boston, Massachusetts (V.Y.R.S., T.F., L.D.L., K.M.S.); Health Science Program and Department of Otolaryngology, University of Brasilia, Brasília, Distrito Federal, Brazil (V.Y.R.S., C.A.P.C.O., F.B.); University of Amsterdam, Amsterdam, the Netherlands (N.A.A.); Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Charlestown, Massachusetts (N.A.A., S.S., X.O.B.); Harvard-MIT Program in Speech and Hearing Bioscience and Technology, Boston, Massachusetts (S.D., K.M.S); Department of Neurosurgery, University of California at San Diego, San Diego, California (F.H.H.).
authors.forename: Konstantina M
authors.initials: KM
authors.lastname: Stankovic
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D000067128:Extracellular Vesicles / Q000378:metabolism*; D005260:Female; D006198:Hair Cells, Auditory / Q000473:pathology; D006319:Hearing Loss, Sensorineural / Q000209:etiology / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D051379:Mice; D008875:Middle Aged; D009464:Neuroma, Acoustic / Q000150:complications / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D013136:Spiral Ganglion / Q000378:metabolism* / Q000473:pathology*
chemical_list: D012313:RNA
keywords: cochlea; extracellular vesicles; sensorineural hearing loss; transwell culture; vestibular schwannoma