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Author Notes:

Corresponding Author: Hanjoong Jo Email: hanjoong.jo@bme.gatech.edu

Corresponding Author: Won Jae Lee Email: lwj@snu.ac.kr

Study design: S.K., I.H.J., W.J.L. and H.J.

Conceived, and designed the experiments: S.K., I.H.J., W.J.L., C.W.K. and H.J.

Performed experiments: S.K., I.H.J., C.W.K. and D.W.K.

Analyzed the data: S.K., I.H.J., C.W.K., D.W.K., W.J.L. and H.J.

Statistical analysis and paper writing: S.K., I.H.J. and H.J.

All authors reviewed the manuscript.

The authors declare no competing financial interests.

Subjects:

Research Funding:

This work was supported by funding from National Institutes of Health grants HL119798, HL113451, HL095070 and HL124879 to HJ. HJ is John and Jan Portman Professor.

This work was supported from National Research Foundation of Korea NRF-2015R1A3A2033475 to W.-J. L.

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • Science & Technology - Other Topics
  • ENDOTHELIAL-CELLS
  • DISTURBED FLOW
  • SHEAR-STRESS
  • IMMUNE-RESPONSE
  • HOST-DEFENSE
  • ACTIVATION
  • ATHEROSCLEROSIS
  • EXPRESSION
  • MELANOGASTER
  • DYSFUNCTION

Functional screening of mammalian mechanosensitive genes using Drosophila RNAi library-Smarcd3/Bap60 is a mechanosensitive pro-inflammatory gene

Tools:

Journal Title:

Scientific Reports

Volume:

Volume 6

Publisher:

, Pages 36461-36461

Type of Work:

Article | Final Publisher PDF

Abstract:

Disturbed blood flow (d-flow) induces atherosclerosis by altering the expression of mechanosensitive genes in the arterial endothelium. Previously, we identified >580 mechanosensitive genes in the mouse arterial endothelium, but their role in endothelial inflammation is incompletely understood. From this set, we obtained 84 Drosophila RNAi lines that silences the target gene under the control of upstream activation sequence (UAS) promoter. These lines were crossed with C564-GAL4 flies expressing GFP under the control of drosomycin promoter, an NF-? B target gene and a marker of pathogen-induced inflammation. Silencing of psmd12 or ERN1 decreased infection-induced drosomycin expression, while Bap60 silencing significantly increased the drosomycin expression. Interestingly, knockdown of Bap60 in adult flies using temperature-inducible Bap60 RNAi (C564 ts -GAL4-Bap60-RNAi) enhanced drosomycin expression upon Gram-positive bacterial challenge but the basal drosomycin expression remained unchanged compared to the control. In the mammalian system, smarcd3 (mammalian ortholog of Bap60) expression was reduced in the human- and mouse aortic endothelial cells exposed to oscillatory shear in vitro as well as in the d-flow regions of mouse arterial endothelium in vivo. Moreover, siRNA-mediated knockdown of smarcd3 induced endothelial inflammation. In summary, we developed an in vivo Drosophila RNAi screening method to identify flow-sensitive genes that regulate endothelial inflammation.

Copyright information:

© The Author(s) 2016.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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