Methods of improving RNAi in well-differentiated airway epithelia
The present invention relates to methods of reducing a level of a target mRNA in a well-differentiated airway epithelial cell by contacting the cell with a facilitating agent followed by contacting the cell with a therapeutic RNAi agent, wherein the facilitating agent comprises a Platelet-activating factor receptor (PAFR) ligand.
1. A method of reducing a level of a target mRNA in a well-differentiated airway epithelial cell comprising contacting the cell with a therapeutic RNAi agent and a composition comprising a Platelet-activating factor receptor (PAFR) ligand, wherein the mRNA level of the target mRNA is reduced by at least 1% as compared to a control cell that has not been contacted with PAFR, wherein the PAFR is Platelet-activating factor (PAF), lipooligosaccharide (LOS), spiperone and/or teichoic acid, wherein the therapeutic RNAi agent is a Dicer-substrate RNA (DsiRNA) molecule comprising SEQ ID NO: 1 and SEQ ID NO: 2 (hsHPRT); SEQ ID NO: 3 and SEQ ID NO: 4 (CFTR); SEQ ID NO: 5 and SEQ ID NO: 6 (RSV N); or SEQ ID NO: 7 and SEQ ID NO: 8 (DIG-HPRT).
2. The method of claim 1 , wherein well-differentiated cell is more than five days old.
3. The method of claim 1 , wherein the airway epithelial cell is a lung cell, a nasal cell, a tracheal cell, a bronchial cell, a bronchiolar or alveolar epithelial cell.
4. The method of claim 1 , wherein the airway epithelial cells are present in a mammal.
5. The method of claim 1 , wherein the airway epithelial cell is a CF epithelial cell.
6. The method of claim 1 , wherein the therapeutic RNAi agent and the composition is administered orally or by inhalation.
7. The method of claim 1 , wherein the PAFR ligand is present within a pharmaceutical composition.
8. A method of treating a subject having an airway epithelial disease comprising administering to the subject an effective amount of a therapeutic agent and a composition comprising a Platelet-activating factor receptor (PAFR) ligand to alleviate the symptoms of the airway epithelial disease by inducing a therapeutic effect, wherein the symptoms are reduced by at least 1%, wherein the therapeutic agent comprises Aminoglutethimide, Biperiden, Diphenhydramine, Rottlerin, Midodrine, Thioridazine, Sulfadimethoxine, neostigmine bromide, Pyridostigmine, pizotifen, tyrophostin (AG-1478), valproic acid, Scriptaid, neomycin, or an RNAi agent that is a Dicer-substrate RNA (DsiRNA) molecule comprising SEQ ID NO: 1 and SEQ ID NO: 2 (hsHPRT); SEQ ID NO: 3 and SEQ ID NO: 4 (CFTR); SEQ ID NO: 5 and SEQ ID NO: 6 (RSV N); or SEQ ID NO: 7 and SEQ ID NO: 8 (DIG-HPRT).
9. The method of claim 8 , wherein the PAFR is Platelet-activating factor (PAF), lipooligosaccharide (LOS), spiperone and/or teichoic acid.
10. The method of claim 8 , wherein the therapeutic RNAi agent and the composition is administered orally or by inhalation.
11. The method of claim 8 , wherein the therapeutic RNAi agent and the composition is administered by aerosol, dry powder, bronchoscopic instillation, or intra-airway (tracheal or bronchial) aerosol and the therapeutic agent is administered orally, by inhalation, by aerosol, dry powder, bronchoscopic instillation, or intra-airway (tracheal or bronchial) aerosol.
12. The method of claim 8 , wherein the therapeutic agent is present within a pharmaceutical composition.
13. The method of claim 8 , wherein the airway epithelial disease is cystic fibrosis.