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

• Published in: Scientific reports Vol. 6; no. 1; p. 36461
• Main Authors: Kumar, Sandeep, Jang, In-hwan, Kim, Chan Woo, Kang, Dong-Won, Lee, Won Jae, Jo, Hanjoong
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.11.2016; Nature Publishing Group
• Subjects: 38/35; 38/89; 38/90; 45; 631/1647; 631/1647/2017; Animals; Aorta; Aorta - cytology; Arteriosclerosis; Blood flow; Cells, Cultured; Chromosomal Proteins, Non-Histone - antagonists & inhibitors; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; Drosomycin; Drosophila - metabolism; Drosophila Proteins - antagonists & inhibitors; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - metabolism; Endothelium; Gene flow; Gene Library; Genes; Human Umbilical Vein Endothelial Cells; Humanities and Social Sciences; Humans; Inflammation; Inflammation - etiology; Insects; Interleukin-8 - metabolism; Mice; Mice, Inbred C57BL; multidisciplinary; Muscle Proteins - antagonists & inhibitors; Muscle Proteins - genetics; Muscle Proteins - metabolism; Promoter Regions, Genetic; RNA Interference; RNA-mediated interference; Science; Shear Strength; siRNA; Temperature effects; Transcription Factors - antagonists & inhibitors; Transcription Factors - genetics; Transcription Factors - metabolism; Vascular Cell Adhesion Molecule-1 - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep36461

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-κB 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.