Adenovirus-Mediated In Vivo Silencing of Anaphylatoxin Receptor C5aR

[摘要] C5a, one of the most potent inflammatory peptides, induces itsinflammatory functions by interacting with C5a receptor (C5aR)that belongs to the rhodopsin family of seven-transmembrane Gprotein-coupled receptors. C5a/C5aR signaling has been implicatedin the pathogenesis of many inflammatory and immunologicaldiseases such as sepsis and acute lung injury. Widespreadupregulation of C5aR has been seen at both the protein level andtranscriptional level under pathological conditions. Here, we showthat C5aR gene expression can be specifically suppressed by siRNA,both in vitro and in vivo. A panel of chemically siRNAoligonucleotides was first synthesized to identify the functionalsiRNA sequences. The short hairpin RNAs (shRNAs) were alsodesigned, cloned, and tested for the silencing effects in C5aRtransfected cells. The effective shRNA expression cassettes werethen transferred to an adenovirus DNA vector. ShRNA-expressingadenoviruses were intratracheally administered into mouse lung,and a significant in vivo silencing of C5aR was obtained four daysafter administration. Thus, C5aR shRNA-expressing adenovirusesappear to be an alternative strategy for the treatment ofcomplement-induced disorders.

[学科分类] 基础医学