title: Lentiviral delivery of RNAi for in vivo lineage-specific modulation of gene expression in mouse lung macrophages.
abstract: Although RNA interference (RNAi) has become a ubiquitous laboratory tool since its discovery 12 years ago, in vivo delivery to selected cell types remains a major technical challenge. Here, we report the use of lentiviral vectors for long-term in vivo delivery of RNAi selectively to resident alveolar macrophages (AMs), key immune effector cells in the lung. We demonstrate the therapeutic potential of this approach by RNAi-based downregulation of p65 (RelA), a component of the pro-inflammatory transcriptional regulator, nuclear factor κB (NF-κB) and a key participant in lung disease pathogenesis. In vivo RNAi delivery results in decreased induction of NF-κB and downstream neutrophilic chemokines in transduced AMs as well as attenuated lung neutrophilia following stimulation with lipopolysaccharide (LPS). Through concurrent delivery of a novel lentiviral reporter vector (lenti-NF-κB-luc-GFP) we track in vivo expression of NF-κB target genes in real time, a critical step towards extending RNAi-based therapy to longstanding lung diseases. Application of this system reveals that resident AMs persist in the airspaces of mice following the resolution of LPS-induced inflammation, thus allowing these localized cells to be used as effective vehicles for prolonged RNAi delivery in disease settings.
authors: Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
authors: Andrew A
authors: AA
authors: Wilson
authors: Letty W
authors: LW
authors: Kwok
authors: Emily L
authors: EL
authors: Porter
authors: Julie G
authors: JG
authors: Payne
authors: Gregory S
authors: GS
authors: McElroy
authors: Sarah J
authors: SJ
authors: Ohle
authors: Sara R
authors: SR
authors: Greenhill
authors: Matthew T
authors: MT
authors: Blahna
authors: Kazuko
authors: K
authors: Yamamoto
authors: Jyh C
authors: JC
authors: Jean
authors: Joseph P
authors: JP
authors: Mizgerd
authors: Darrell N
authors: DN
authors: Kotton
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology; D016086:Lentivirus / Q000235:genetics*; D008168:Lung / Q000276:immunology* / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D016676:Macrophages, Alveolar / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000235:genetics; D034622:RNA Interference / Q000502:physiology; D051996:Transcription Factor RelA / Q000235:genetics
chemical_list: D016328:NF-kappa B; D051996:Transcription Factor RelA
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