Antisense therapy against CCR3 and the common beta chain attenuates allergen-induced eosinophilic responses.
BACKGROUND
The drug product TPI ASM8 contains two modified phosphorothioate antisense oligonucleotides designed to inhibit allergic inflammation by down-regulating human CCR3 and the common beta chain (beta(c)) of IL-3, IL-5, and granulocyte-macrophage colony-stimulating factor receptors.


OBJECTIVE
This study examined the effects of inhaled TPI ASM8 on sputum cellular influx, CCR3 and beta(c) mRNA and protein levels, and the airway physiologic response after inhaled allergen.


METHODS
Seventeen subjects with mild atopic asthma were randomized in a crossover study to inhale 1,500 microg TPI ASM8 or placebo by nebulizer, once daily for 4 days. On Day 3, subjects underwent allergen inhalation challenge. Sputum samples were collected before and after allergen. CCR3 and beta(c) protein levels were measured by flow cytometry, mRNA was measured using real-time quantitative polymerase chain reaction, and the FEV1 was measured over 7 hours after challenge.


RESULTS
Compared with placebo, TPI ASM8 inhibited sputum eosinophil influx by 46% (P = 0.02) and blunted the increase in total cells (63%) after allergen challenge. TPI ASM8 significantly reduced the early asthmatic response (P = 0.04) with a trend for the late asthmatic response (P = 0.08). The allergen-induced (Day 2 to Day 3) levels of beta(c) mRNA and CCR3 mRNA in sputum-derived cells were inhibited by TPI ASM8 (P = 0.039 and P = 0.054, respectively), with no significant effects on the cell surface protein expression of CCR3 and beta(c) (P > 0.05). No serious adverse events were reported.


CONCLUSIONS
TPI ASM8 attenuates the allergen-induced increase in target gene mRNA and airway responses in subjects with mild asthma. Clinical trial registered with www.clinicaltrials.gov (NCT 00264966).
Gauvreau
Gail M
GM
Department of Medicine, McMaster University, Hamilton, Ontario, Canada. gauvreau@mcmaster.ca
Boulet
Louis Philippe
LP
Cockcroft
Donald W
DW
Baatjes
Adrian
A
Cote
Johanne
J
Deschesnes
Francine
F
Davis
Beth
B
Strinich
Tara
T
Howie
Karen
K
Duong
Mylinh
M
Watson
Richard M
RM
Renzi
Paolo M
PM
O'Byrne
Paul M
PM
D000280:Administration, Inhalation; D000328:Adult; D000485:Allergens / Q000009:adverse effects*; D001249:Asthma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D018592:Cross-Over Studies; D004311:Double-Blind Method; D004338:Drug Combinations; D005260:Female; D005434:Flow Cytometry; D005500:Follow-Up Studies; D005541:Forced Expiratory Volume; D015870:Gene Expression; D006801:Humans; D008297:Male; D008875:Middle Aged; D009330:Nebulizers and Vaporizers; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000627:therapeutic use*; D054735:Phosphorothioate Oligonucleotides / Q000008:administration & dosage / Q000627:therapeutic use*; D011657:Pulmonary Eosinophilia / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D054397:Receptors, CCR3 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D018121:Receptors, Cytokine / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013183:Sputum / Q000166:cytology / Q000378:metabolism; D016896:Treatment Outcome
D000485:Allergens; C515350:CCR3 protein, human; D004338:Drug Combinations; D016376:Oligonucleotides, Antisense; D054735:Phosphorothioate Oligonucleotides; D012333:RNA, Messenger; D054397:Receptors, CCR3; D018121:Receptors, Cytokine; C559988:TPI ASM8; C114668:beta C receptor