title: TGFβ Triggers miR-143/145 Transfer From Smooth Muscle Cells to Endothelial Cells, Thereby Modulating Vessel Stabilization.
abstract: BACKGROUND
  The miR-143/145 cluster is highly expressed in smooth muscle cells (SMCs), where it regulates phenotypic switch and vascular homeostasis. Whether it plays a role in neighboring endothelial cells (ECs) is still unknown.
  
  
  OBJECTIVE
  To determine whether SMCs control EC functions through passage of miR-143 and miR-145.
  
  
  RESULTS
  We used cocultures of SMCs and ECs under different conditions, as well as intact vessels to assess the transfer of miR-143 and miR-145 from one cell type to another. Imaging of cocultured cells transduced with fluorescent miRNAs suggested that miRNA transfer involves membrane protrusions known as tunneling nanotubes. Furthermore, we show that miRNA passage is modulated by the transforming growth factor (TGF) β pathway because both a specific transforming growth factor-β (TGFβ) inhibitor (SB431542) and an shRNA against TGFβRII suppressed the passage of miR-143/145 from SMCs to ECs. Moreover, miR-143 and miR-145 modulated angiogenesis by reducing the proliferation index of ECs and their capacity to form vessel-like structures when cultured on matrigel. We also identified hexokinase II (HKII) and integrin β 8 (ITGβ8)-2 genes essential for the angiogenic potential of ECs-as targets of miR-143 and miR-145, respectively. The inhibition of these genes modulated EC phenotype, similarly to miR-143 and miR-145 overexpression in ECs. These findings were confirmed by ex vivo and in vivo approaches, in which it was shown that TGFβ and vessel stress, respectively, triggered miR-143/145 transfer from SMCs to ECs.
  
  
  CONCLUSIONS
  Our results demonstrate that miR-143 and miR-145 act as communication molecules between SMCs and ECs to modulate the angiogenic and vessel stabilization properties of ECs.
authors: From IRCCS MultiMedica, Milan, Italy (M.C.); Humanitas Clinical and Research Center, Rozzano (MI), Italy (M.Q., M.M., G.C., L.E.); Milan Unit of the Institute of Genetic and Biomedical Research, Rozzano (MI), Italy (G.C., L.E.); Department of Cardiovascular Diseases, University of Milan, Rozzano (MI), Italy (G.C.); Department of Clinical and Experimental Medicine, Center of Excellence for Toxicological Research (CERT), University of Parma, Parma, Italy (M.M.); and Department of Medicine, University of California, San Diego (J.C.).
authors: Montserrat
authors: M
authors: Climent
authors: From IRCCS MultiMedica, Milan, Italy (M.C.); Humanitas Clinical and Research Center, Rozzano (MI), Italy (M.Q., M.M., G.C., L.E.); Milan Unit of the Institute of Genetic and Biomedical Research, Rozzano (MI), Italy (G.C., L.E.); Department of Cardiovascular Diseases, University of Milan, Rozzano (MI), Italy (G.C.); Department of Clinical and Experimental Medicine, Center of Excellence for Toxicological Research (CERT), University of Parma, Parma, Italy (M.M.); and Department of Medicine, University of California, San Diego (J.C.).
authors: Manuela
authors: M
authors: Quintavalle
authors: From IRCCS MultiMedica, Milan, Italy (M.C.); Humanitas Clinical and Research Center, Rozzano (MI), Italy (M.Q., M.M., G.C., L.E.); Milan Unit of the Institute of Genetic and Biomedical Research, Rozzano (MI), Italy (G.C., L.E.); Department of Cardiovascular Diseases, University of Milan, Rozzano (MI), Italy (G.C.); Department of Clinical and Experimental Medicine, Center of Excellence for Toxicological Research (CERT), University of Parma, Parma, Italy (M.M.); and Department of Medicine, University of California, San Diego (J.C.).
authors: Michele
authors: M
authors: Miragoli
authors: From IRCCS MultiMedica, Milan, Italy (M.C.); Humanitas Clinical and Research Center, Rozzano (MI), Italy (M.Q., M.M., G.C., L.E.); Milan Unit of the Institute of Genetic and Biomedical Research, Rozzano (MI), Italy (G.C., L.E.); Department of Cardiovascular Diseases, University of Milan, Rozzano (MI), Italy (G.C.); Department of Clinical and Experimental Medicine, Center of Excellence for Toxicological Research (CERT), University of Parma, Parma, Italy (M.M.); and Department of Medicine, University of California, San Diego (J.C.).
authors: Ju
authors: J
authors: Chen
authors: From IRCCS MultiMedica, Milan, Italy (M.C.); Humanitas Clinical and Research Center, Rozzano (MI), Italy (M.Q., M.M., G.C., L.E.); Milan Unit of the Institute of Genetic and Biomedical Research, Rozzano (MI), Italy (G.C., L.E.); Department of Cardiovascular Diseases, University of Milan, Rozzano (MI), Italy (G.C.); Department of Clinical and Experimental Medicine, Center of Excellence for Toxicological Research (CERT), University of Parma, Parma, Italy (M.M.); and Department of Medicine, University of California, San Diego (J.C.). gianluigi.condorelli@humanitasresearch.it leonardo.elia@yahoo.com.
authors: Gianluigi
authors: G
authors: Condorelli
authors: From IRCCS MultiMedica, Milan, Italy (M.C.); Humanitas Clinical and Research Center, Rozzano (MI), Italy (M.Q., M.M., G.C., L.E.); Milan Unit of the Institute of Genetic and Biomedical Research, Rozzano (MI), Italy (G.C., L.E.); Department of Cardiovascular Diseases, University of Milan, Rozzano (MI), Italy (G.C.); Department of Clinical and Experimental Medicine, Center of Excellence for Toxicological Research (CERT), University of Parma, Parma, Italy (M.M.); and Department of Medicine, University of California, San Diego (J.C.). gianluigi.condorelli@humanitasresearch.it leonardo.elia@yahoo.com.
authors: Leonardo
authors: L
authors: Elia
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D001808:Blood Vessels / Q000378:metabolism / Q000502:physiology; D015153:Blotting, Western; D002450:Cell Communication / Q000187:drug effects / Q000235:genetics; D022081:Cell Surface Extensions / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D004149:Dioxoles / Q000494:pharmacology; D005786:Gene Expression Regulation; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D039641:Integrin beta Chains / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D032389:Myocytes, Smooth Muscle / Q000378:metabolism* / Q000648:ultrastructure; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; D039641:Integrin beta Chains; C543499:MIRN145a microRNA, mouse; D035683:MicroRNAs; C532884:MIRN143 microRNA, mouse; D016212:Transforming Growth Factor beta; C094865:integrin beta8; D006593:Hexokinase
keywords: angiogenesis effect; endothelial cells; microRNAs; smooth muscle myocytes