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

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-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 (C564ts-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.

Identify Candidate Genes in the Interaction between Abdominal Aortic Aneurysm and Type 2 Diabetes Mellitus by Using Biomedical Discovery Support System

“…[35] Research found that the inducible knockdown of RPN5 had a vital inhibitory effect on Toll and IMD-mediated NF-κB activation, so Rpn5 might be a mechanosensitive NF-κB activator. [36] Study also revealed that the expression of osteopontin and NF-κB elevated in human aortic aneurysms, which accelerate the degradation of the extracellular matrix and play a significant role in pathogenesis and development of TAA and AAA. [37] In addition, intracellular NF-κB signaling played an important role in remodeling of blood vessels and formation of aneurysms.…”

LncRNA SMARCD3-OT1 Promotes Muscle Hypertrophy and Fast-Twitch Fiber Transformation via Enhancing SMARCD3X4 Expression

“…The protein encoded by the SMARCD3 gene is a member of the SWI/SNF protein family. Its members have helicase and ATPase activities and can regulate the transcription of genes by changing the chromatin structure around the genes, playing an important role in physiological processes [13][14][15]. Reports show that SMARCD3 can regulate the cell cycle through the CDKN1A signaling pathway, affecting tumor formation [16], and it can also regulate muscle development in zebrafish through MYOD and MYF5 signaling pathways [17,18].…”

Identification of phenotype-specific networks from paired gene expression–cell shape imaging data

“…Cell morphology and tumour organisation have been found to be a factor in modulating the intracellular signalling state through pathways able to integrate mechanical stimuli from the extracellular environment (Miralles et al, 2003;Olson and Nordheim, 2010;Orsulic et al, 1999;Zheng et al, 2009). The discovery of mechanosensitive pathways in various tissues has revealed a complex interplay between cell morphology and signalling (Kumar et al, 2016). Further studies have revealed that cell morphology can also be a predictor of tumorigenic and metastatic potential as certain nuclear and cytoplasmic features enhance cell motility and spread to secondary sites (Wu et al, 2020), aided by the Epithelial to Mesenchymal Transition (EMT).…”