title: SASH1, a new potential link between smoking and atherosclerosis.
abstract: OBJECTIVE
  We have previously reported that SASH1 expression is increased in circulating human monocytes from smokers and was positively correlated with the number of carotid atherosclerotic plaques. The aim of this study was to further validate the link between smoking, SASH1 and atherosclerosis within the vascular wall and to assess the impact of SASH1 expression on endothelial cell functions.
  
  
  METHODS
  Human carotids with atherosclerotic plaques were obtained from 58 patients (45 of them with known smoking status: smoker, non-smoker, ex-smokers), and were processed for gene expression analyses and immunostaining. To investigate its function, SASH1 was silenced in human aortic endothelial cells (HAECs) using two different siRNA and subcellular localization of SASH1 was determined by immunostaining and subcellular fractionation. Subsequently the transcriptomic analyses and functional experiments (wound healing, WST-1 proliferation or Matrigel assays) were performed to characterize SASH1 function.
  
  
  RESULTS
  SASH1 was expressed in human vascular cells (HAECs, smooth muscle cells) and in monocytes/macrophages. Its tissue expression was significantly higher in the atherosclerotic carotids of smokers compared to non-smokers (p < 0.01). In HAECs, SASH1 was expressed mostly in the cytoplasm and SASH1 knockdown resulted in an increased cell migration, proliferation and angiogenesis. Transcriptomic and pathway analyses showed that SASH1 silencing results in a decreased CYP1A1 expression possibly through the inhibition of TP53 activity.
  
  
  CONCLUSIONS
  We showed that SASH1 expression is increased in atherosclerotic carotids in smokers and its silencing affects endothelial angiogenic functions; therefore we provide a potential link between smoking and atherosclerosis through SASH1 expression.
authors: Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France; University Heart Center Hamburg, Clinic for General and Interventional Cardiology, Hamburg, Germany; German Center for Cardiovascular Research (DZHK e.V.) Partner Site Hamburg, Lübeck, Kiel, Hamburg, Germany. Electronic address: h.weide@hotmail.fr.
authors: Henri
authors: H
authors: Weidmann
authors: Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France.
authors: Zahia
authors: Z
authors: Touat-Hamici
authors: Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France.
authors: Herve
authors: H
authors: Durand
authors: University Heart Center Hamburg, Clinic for General and Interventional Cardiology, Hamburg, Germany; German Center for Cardiovascular Research (DZHK e.V.) Partner Site Hamburg, Lübeck, Kiel, Hamburg, Germany.
authors: Christian
authors: C
authors: Mueller
authors: Sorbonne Universités, UPMC, UMS_2 Omique, Plateforme P3S, F-75005, Paris, France; INSERM, UMS 29 Omique, Plateforme P3S, F-75005, Paris, France.
authors: Solenne
authors: S
authors: Chardonnet
authors: Sorbonne Universités, UPMC, UMS_2 Omique, Plateforme P3S, F-75005, Paris, France; INSERM, UMS 29 Omique, Plateforme P3S, F-75005, Paris, France.
authors: Cedric
authors: C
authors: Pionneau
authors: Department of Pathology, Hôpital de la Pitié-Salpêtrière, AP-HP, Paris, France.
authors: Frédéric
authors: F
authors: Charlotte
authors: Department of Surgery, Klinikum rechts der Isar, Technische Universität München, IsmaningerStrasse 22, Munich, Germany.
authors: Klaus-Peter
authors: KP
authors: Janssen
authors: Programa de Genetica Humana ICBM, Facultad de Medicina, Universidad de Chile, Chile.
authors: Ricardo
authors: R
authors: Verdugo
authors: Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France.
authors: Francois
authors: F
authors: Cambien
authors: University Heart Center Hamburg, Clinic for General and Interventional Cardiology, Hamburg, Germany; German Center for Cardiovascular Research (DZHK e.V.) Partner Site Hamburg, Lübeck, Kiel, Hamburg, Germany.
authors: Stefan
authors: S
authors: Blankenberg
authors: Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France.
authors: Laurence
authors: L
authors: Tiret
authors: University Heart Center Hamburg, Clinic for General and Interventional Cardiology, Hamburg, Germany; German Center for Cardiovascular Research (DZHK e.V.) Partner Site Hamburg, Lübeck, Kiel, Hamburg, Germany.
authors: Tanja
authors: T
authors: Zeller
authors: Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France. Electronic address: ewa.ninio@upmc.fr.
authors: Ewa
authors: E
authors: Ninio
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001011:Aorta / Q000378:metabolism; D050197:Atherosclerosis / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D002453:Cell Cycle; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000378:metabolism; D056743:Cyclin D3 / Q000378:metabolism; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D020868:Gene Silencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D009389:Neovascularization, Pathologic; D034741:RNA, Small Interfering / Q000378:metabolism; D012907:Smoking / Q000378:metabolism*; D059467:Transcriptome; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
chemical_list: C530104:CCND1 protein, human; C534696:CCND3 protein, human; D056743:Cyclin D3; D034741:RNA, Small Interfering; C474743:SASH1 protein, human; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D019938:Cyclin D1; C569463:CYP1A1 protein, human; D019363:Cytochrome P-450 CYP1A1
keywords: Atherosclerosis; CCND1; CCND3; SASH1; Smoking; TP53