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A novel function for cadherin 11/osteoblast-cadherin in vascular smooth muscle cells: Modulation of cell migration and proliferation  Thomas S. Monahan,

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Presentation on theme: "A novel function for cadherin 11/osteoblast-cadherin in vascular smooth muscle cells: Modulation of cell migration and proliferation  Thomas S. Monahan,"— Presentation transcript:

1 A novel function for cadherin 11/osteoblast-cadherin in vascular smooth muscle cells: Modulation of cell migration and proliferation  Thomas S. Monahan, MD, Nicholas D. Andersen, BS, Haig Panossian, BA, Jeffrey A. Kalish, MD, Soizic Daniel, PhD, Gautam V. Shrikhande, MD, Christiane Ferran, MD, PhD, Frank W. LoGerfo, MD  Journal of Vascular Surgery  Volume 45, Issue 3, Pages (March 2007) DOI: /j.jvs Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

2 Fig 1 Vein grafts demonstrate increased expression of cadherin 11/osteoblast-cadherin (CDH11). Vein grafts recovered after implantation in a canine cephalic vein interposition graft model. (A) Vein grafts harvested after 1, (B) 7, (C) 14, and (D) 30 days of implantation were stained for CDH11. All sections are oriented with the lumen of the graft to the right of the panel. The formation of intimal hyperplasia (IH) is appreciable at 7 days. (E) A section of control, nonimplanted vein is presented for comparison. All images are at original magnification ×10. Note that the intensity of staining for CDH11 (arrows) is increased in the specimens retrieved at 7, 14, and 30 days. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

3 Fig 2 Tumor necrosis factor-α (TNF-α) increases osteoblast-cadherin/cadherin 11 (CDH11) messenger RNA in cultured vascular smooth muscle cells. Cells were continually stimulated with 10 ng/mL of TNF-α and recovered at predetermined time points. CDH11 transcription reached its maximum of 1.7 times greater than baseline at 8 hours. Data are presented as mean ± standard deviation. †enotes P < .01. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

4 Fig 3 Inhibition of osteoblast (OB)-cadherin/cadherin 11 (CDH11), which attenuates vascular smooth muscle migration. A, The decrease in migration achieved with the CDH11-blocking antibody was dose dependent. Neutral control antibody did not effect smooth muscle cell migration. The trend in attenuation is statistically significant by linear regression (P = .03). B, Attenuation of cellular migration is also observed after transfection with small interfering (siRNA) targeting CDH11. Transfection of vascular smooth muscle cells with siRNA decreased cellular migration to 67% of untreated cells (P = .036). Data are presented as mean ± standard deviation.*Denotes P < .05. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

5 Fig 4 Silencing RNA–mediated knockdown of osteoblast-cadherin/cadherin (CDH11) in vascular smooth muscle cells. A, Gene silencing caused a time-dependent and dose-dependent decrease in CDH11 protein. Protein content was assayed 2, 4, and 6 days after transfection. NT, Cells that received no treatment; Con, cells treated with control or control small interfering RNA (siRNA); CDH, represents cells treated with siRNA targeting CDH11; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Maximal protein knockdown occurred after 6 days. The degree of protein knockdown was dose-dependent. B, Transfection with 50 nM and 100 nM of siRNA targeting CDH11 decreases cellular messenger levels by 54% ± 5% and 69% ± 4%, respectively, compared with cells treated with control siRNA. Data are presented as mean ± standard deviation. CDH11 mRNA relative to 18s mRNA normalized to cells treated with control siRNA. *Denotes P < .05. †Denotes P < .01. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

6 Fig 5 Inhibition of osteoblast (OB)-cadherin/cadherin 11 (CDH11) attenuates vascular smooth muscle cell proliferation. A, Cellular proliferation was measured immediately before the addition of the OB-cadherin–blocking antibody, then every 2 days thereafter by the Alamar Blue assay. Proliferation was normalized to the emission value obtained on day 0. Neutral control antibody did not effect SMC proliferation. B, CDH11 knockdown mediated by silencing RNA also attenuates vascular SMC proliferation. Attenuation of proliferation was observed at a later time point in cells treated with small interfering RNA (siRNA). This delay likely reflects the delay observed in protein knockdown. Data are presented as mean ± standard deviation. *Denotes P < .05. †Denotes P < .01. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

7 Fig 6 Fluorescence-activated cell-sorting analysis of cells treated with the osteoblast-cadherin/cadherin 11 (CDH11)–blocking antibody (Ab). There is no difference in the proportion of apoptotic cells (A0) or cells in S-phase in both treatment groups. Cells treated with the CDH11-blocking antibody had a greater proportion in the G1/G0 phase of the cell cycle (P = .003) and lesser proportion in the G2/M phase of the cell cycle (P < .001). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2007 The Society for Vascular Surgery Terms and Conditions

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