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Shear stress-induced transcriptional regulation via hybrid promoters as a potential tool for promoting angiogenesis

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Abstract

Among the key effects of fluid shear stress on vascular endothelial cells is modulation of gene expression. Promoter sequences termed shear stress response elements (SSREs) mediate the responsiveness of endothelial genes to shear stress. While previous studies showed that shear stress responsiveness is mediated by a single SSRE, these endogenous promoters often encode for multiple SSREs. Moreover, hybrid promoters encoding a single SSRE rarely respond to shear stress at the same magnitude as the endogenous promoter. Thus, to better understand the interplay between the various SSREs, and between SSREs and endothelial-specific sequences (ESS), we generated a series of constructs regulated by SSREs cassettes alone, or in combination with ESS, and tested their response to shear stress and endothelial specific expression. Among these constructs, the most responsive promoter (NR1/2) encoded a combination of two GAGACC/SSREs, the Sp1/Egr1 sequence, as well as a TPA response element (TRE). This construct was four- to five-fold more responsive to shear stress than a promoter encoding a single SSRE. The expression of constructs containing other SSRE combinations was unaffected or suppressed by shear stress. Addition of ESS derived from the Tie2 promoter, either 5′ or 3′ to NR1/2 resulted in shear stress transcriptional suppression, yet retained endothelial specific expression. Thus, the combination and localization order of the various SSREs in a single promoter is crucial in determining the pattern and degree of shear stress responsiveness. These shear stress responsive cassettes may prove beneficial in our attempt to time the expression of an endothelial transgene in the vasculature.

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Acknowledgments

We would like to express our thanks to Dr. E. Sabo (Department of Pathology, Carmel Medical Center, Haifa, Israel) for his experienced input on the statistical analysis of the data. This study was supported by the Israeli Academy of Sciences (436-02, OB, NR), The Israeli Ministry of Health (4988, OB, NR), the Israeli Science Ministry 01-299-01-01 (NR, SE), the Rappaport Family Institute for Research in the Medical Sciences.

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Authors and Affiliations

  1. Department of Physiology, Ruth & Bruce Rappaport Faculty of Medicine, P.O. Box 9649, Bat Galim, Haifa, 31096, Israel

    Michal Silberman, Yaron D. Barac & Ofer Binah

  2. Rappaport Family Institute for Research in the Medical Sciences, Haifa, Israel

    Michal Silberman, Yaron D. Barac & Ofer Binah

  3. Department Anatomy and Cell Biology, Ruth & Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel

    Hava Yahav & Efrat Wolfovitz

  4. Department of Biomedical Engineering, Tel-Aviv University, Tel-Aviv, Israel

    Shmuel Einav

  5. The Vascular Center for Excellence, Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA

    Nitzan Resnick

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  1. Michal Silberman
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  3. Hava Yahav
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Correspondence to Ofer Binah.

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Michal Silberman and Yaron D. Barac have contributed equally to this manuscript.

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Silberman, M., Barac, Y.D., Yahav, H. et al. Shear stress-induced transcriptional regulation via hybrid promoters as a potential tool for promoting angiogenesis. Angiogenesis 12, 231–242 (2009). https://doi.org/10.1007/s10456-009-9143-7

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