Journal article
Platelet Activating Factor Receptor Activation Improves siRNA Uptake and RNAi Responses in Well-differentiated Airway Epithelia
Molecular therapy. Nucleic acids, Vol.3(7), pp.e175-e175
07/2014
DOI: 10.1038/mtna.2014.26
PMCID: PMC4121516
PMID: 25025465
Abstract
Well-differentiated human airway epithelia present formidable barriers to efficient siRNA delivery. We previously reported that treatment of airway epithelia with specific small molecules improves oligonucleotide uptake and facilitates RNAi responses. Here, we exploited the platelet activating factor receptor (PAFR) pathway, utilized by specific bacteria to transcytose into epithelia, as a trigger for internalization of Dicer-substrate siRNAs (DsiRNA). PAFR is a G-protein coupled receptor which can be engaged and activated by phosphorylcholine residues on the lipooligosaccharide (LOS) of nontypeable
Haemophilus influenzae
and the teichoic acid of
Streptococcus pneumoniae
as well as by its natural ligand, platelet activating factor (PAF). When well-differentiated airway epithelia were simultaneously treated with either nontypeable
Haemophilus influenzae
LOS or PAF and transduced with DsiRNA formulated with the peptide transductin, we observed silencing of both endogenous and exogenous targets. PAF receptor antagonists prevented LOS or PAF-assisted DsiRNA silencing, demonstrating that ligand engagement of PAFR is essential for this process. Additionally, PAF-assisted DsiRNA transfection decreased CFTR protein expression and function and reduced exogenous viral protein levels and titer in human airway epithelia. Treatment with spiperone, a small molecule identified using the Connectivity map database to correlate gene expression changes in response to drug treatment with those associated with PAFR stimulation, also induced silencing. These results suggest that the signaling pathway activated by PAFR binding can be manipulated to facilitate siRNA entry and function in difficult to transfect well-differentiated airway epithelial cells.
Details
- Title: Subtitle
- Platelet Activating Factor Receptor Activation Improves siRNA Uptake and RNAi Responses in Well-differentiated Airway Epithelia
- Creators
- Sateesh Krishnamurthy - , Iowa City, IowaMark A Behlke - , Coralville, IowaMichael A Apicella - , Iowa City, IowaPaul B McCray - , Iowa City, IowaBeverly L Davidson - , Iowa City, Iowa
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Nucleic acids, Vol.3(7), pp.e175-e175
- DOI
- 10.1038/mtna.2014.26
- PMID
- 25025465
- PMCID
- PMC4121516
- NLM abbreviation
- Mol Ther Nucleic Acids
- ISSN
- 2162-2531
- eISSN
- 2162-2531
- Publisher
- Nature Publishing Group
- Language
- English
- Date published
- 07/2014
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine
- Record Identifier
- 9984093340002771
Metrics
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