A multi-target antisense approach against PDE4 and PDE7 reduces smoke-induced lung inflammation in mice.
BACKGROUND
Recent development in the field of COPD has focused on strategies aimed at reducing the underlying inflammation through selective inhibition of the phosphodiesterase type IV (PDE4) isoform. Although the anti-inflammatory and bronchodilator activity of selective PDE4 inhibitors has been well documented, their low therapeutic ratio and dose-dependent systemic side effects have limited their clinical utility. This study examined the effect of 2'-deoxy-2'-Fluoro-beta-D-Arabinonucleic Acid (FANA)-containing antisense oligonucleotides (AON) targeting the mRNA for the PDE4B/4D and 7A subtypes on lung inflammatory markers, both in vitro and in vivo.


METHODS
Normal human bronchial epithelial (NHBE) cells were transfected with FANA AON against PDE4B/4D and 7A alone or in combination. mRNA levels for target PDE subtypes, as well as secretion of pro-inflammatory chemokines were then measured following cell stimulation. Mice were treated with combined PDE4B/4D and 7A AON via endo-tracheal delivery, or with roflumilast via oral delivery, and exposed to cigarette smoke for one week. Target mRNA inhibition, as well as influx of inflammatory cells and mediators were measured in lung lavages. A two-week smoke exposure protocol was also used to test the longer term potency of PDE4B/4D and 7A AONs.


RESULTS
In NHBE cells, PDE4B/4D and 7A AONs dose-dependently and specifically inhibited expression of their respective target mRNA. When used in combination, PDE4B/4D and 7A AONs significantly abrogated the cytokine-induced secretion of IL-8 and MCP-1 to near baseline levels. In mice treated with combined PDE4B/4D and 7A AONs and exposed to cigarette smoke, significant protection against the smoke-induced recruitment of neutrophils and production of KC and pro-MMP-9 was obtained, which was correlated with inhibition of target mRNA in cells from lung lavages. In this model, PDE AONs exerted more potent and broader anti-inflammatory effects against smoke-induced lung inflammation than roflumilast. Moreover, the protective effect of PDE4B/4D and 7A AON was maintained when a once-weekly treatment schedule was used.


CONCLUSIONS
These results indicate that inhaled AON against PDE4B/4D and 7A have unique effects on biomarkers that are believed to be important in the pathophysiology of COPD, which supports further development as a potential therapy in this disease.
Fortin
Marylène
M
Topigen Pharmaceuticals Inc, Quebec, Canada. marylene.fortin@topigen.com
D'Anjou
Hélène
H
Higgins
Marie-Eve
ME
Gougeon
Jasmine
J
Aubé
Paméla
P
Moktefi
Kamel
K
Mouissi
Sonia
S
Séguin
Serge
S
Séguin
Rosanne
R
Renzi
Paolo M
PM
Paquet
Luc
L
Ferrari
Nicolay
N
D000818:Animals; D001088:Arabinonucleotides / Q000494:pharmacology*; D001980:Bronchi / Q000187:drug effects / Q000201:enzymology; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000187:drug effects / Q000235:genetics*; D054708:Cyclic Nucleotide Phosphodiesterases, Type 7 / Q000187:drug effects / Q000235:genetics*; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D007249:Inflammation / Q000201:enzymology / Q000503:physiopathology*; D007371:Interferon-gamma / Q000494:pharmacology; D053583:Interleukin-1beta / Q000494:pharmacology; D008168:Lung / Q000201:enzymology / Q000503:physiopathology*; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D012906:Smoke / Q000009:adverse effects*; D012907:Smoking / Q000009:adverse effects*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C507672:2'-deoxy-2'-fluoro-beta-D-arabinonucleic acid; D001088:Arabinonucleotides; D053583:Interleukin-1beta; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D012906:Smoke; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D054708:Cyclic Nucleotide Phosphodiesterases, Type 7