title: Antisense therapy against CCR3 and the common beta chain attenuates allergen-induced eosinophilic responses.
abstract: 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).
authors.affiliation: Department of Medicine, McMaster University, Hamilton, Ontario, Canada. gauvreau@mcmaster.ca
authors.forename: Gail M
authors.initials: GM
authors.lastname: Gauvreau
authors.forename: Louis Philippe
authors.initials: LP
authors.lastname: Boulet
authors.forename: Donald W
authors.initials: DW
authors.lastname: Cockcroft
authors.forename: Adrian
authors.initials: A
authors.lastname: Baatjes
authors.forename: Johanne
authors.initials: J
authors.lastname: Cote
authors.forename: Francine
authors.initials: F
authors.lastname: Deschesnes
authors.forename: Beth
authors.initials: B
authors.lastname: Davis
authors.forename: Tara
authors.initials: T
authors.lastname: Strinich
authors.forename: Karen
authors.initials: K
authors.lastname: Howie
authors.forename: Mylinh
authors.initials: M
authors.lastname: Duong
authors.forename: Richard M
authors.initials: RM
authors.lastname: Watson
authors.forename: Paolo M
authors.initials: PM
authors.lastname: Renzi
authors.forename: Paul M
authors.initials: PM
authors.lastname: O'Byrne
mesh_terms: 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
chemical_list: 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