title: Biliary exosomes influence cholangiocyte regulatory mechanisms and proliferation through interaction with primary cilia.
abstract: Exosomes are small extracellular vesicles that are thought to participate in intercellular communication. Recent work from our laboratory suggests that, in normal and cystic liver, exosome-like vesicles accumulate in the lumen of intrahepatic bile ducts, presumably interacting with cholangiocyte cilia. However, direct evidence for exosome-ciliary interaction is limited and the physiological relevance of such interaction remains unknown. Thus, in this study, we tested the hypothesis that biliary exosomes are involved in intercellular communication by interacting with cholangiocyte cilia and inducing intracellular signaling and functional responses. Exosomes were isolated from rat bile by differential ultracentrifugation and characterized by scanning, transmission, and immunoelectron microscopy. The exosome-ciliary interaction and its effects on ERK1/2 signaling, expression of the microRNA, miR-15A, and cholangiocyte proliferation were studied on ciliated and deciliated cultured normal rat cholangiocytes. Our results show that bile contains vesicles identified as exosomes by their size, characteristic "saucer-shaped" morphology, and specific markers, CD63 and Tsg101. When NRCs were exposed to isolated biliary exosomes, the exosomes attached to cilia, inducing a decrease of the phosphorylated-to-total ERK1/2 ratio, an increase of miR-15A expression, and a decrease of cholangiocyte proliferation. All these effects of biliary exosomes were abolished by the pharmacological removal of cholangiocyte cilia. Our findings suggest that bile contains exosomes functioning as signaling nanovesicles and influencing intracellular regulatory mechanisms and cholangiocyte proliferation through interaction with primary cilia.
authors.affiliation: Miles and Shirley Fiterman Center for Digestive Diseases, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
authors.forename: Anatoliy I
authors.initials: AI
authors.lastname: Masyuk
authors.forename: Bing Q
authors.initials: BQ
authors.lastname: Huang
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Ward
authors.forename: Sergio A
authors.initials: SA
authors.lastname: Gradilone
authors.forename: Jesus M
authors.initials: JM
authors.lastname: Banales
authors.forename: Tatyana V
authors.initials: TV
authors.lastname: Masyuk
authors.forename: Brynn
authors.initials: B
authors.lastname: Radtke
authors.forename: Patrick L
authors.initials: PL
authors.lastname: Splinter
authors.forename: Nicholas F
authors.initials: NF
authors.lastname: LaRusso
mesh_terms: D000818:Animals; D001659:Biliary Tract / Q000166:cytology*; D049109:Cell Proliferation; D002923:Cilia / Q000502:physiology*; D055354:Exosomes / Q000502:physiology*; D005704:Gallbladder / Q000166:cytology*; D015870:Gene Expression; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D050396:TRPP Cation Channels / Q000235:genetics / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D011956:Receptors, Cell Surface; D050396:TRPP Cation Channels; C484365:Pkhd1 protein, mouse