3091 Association Analysis of Member RAS Oncogene Family Gene Polymorphisms with Aspirin Intolerance in Asthmatic Patients

Tuesday, 6 December 2011
Poster Hall (Cancún Center)

Myung ok Kim , Genome research center for allergy and respiratory disease, buchun, South Korea

Choon-Sik Park , Respiratory and allergy, Soonchunhyang University Bucheon Hospital, Bucheon, South Korea

Tae-Hoon Kim , Soonchunhyang University Hospital Genome Research Center for Allergy and Respiratory Disease, Bucheon, South Korea

Background: Member RAS oncogene family (RAB1A) converts small G protein to rab1 protein , a inflammation of blood eosinophils. Thus, functional alterations of the RAB1A gene may contribute to Aspirin exacerbated respiratory disease (AERD).

Methods:

Asthmatics (n=1277) were categorized into Aspirin exacerbated respiratory disease (AERD) and aspirin-tolerant asthma (ATA). 8 SNPs were genotyped. Messenger RNA expression of the RAB1A gene by peripheral blood mononuclear cell is measured by Real time PCR and reverse transcriptase polymerase chain reaction (RT-PCR). Human PBMC culture supernatant expression of 11-dehydrothromboxane B2. Protein expression of the RAB1A gene by PBMC is measured by RNAi (Knock down) analysis.

Results: The logistic regression analysis showed that the rare allele frequency of +41170 C>G on intron 5 was significantly lower in the AERD group (n=261) than in the ATA group (n=1016) (P=0.002). The linear regression analysis revealed a strong association of +41170 C>G with the aspirin challenge induced-FEV1 fall (P=0.00008). RT-PCR and real time PCR  revealed an exon-4-deleted variants. The level of full-length RAB1A mRNA did not differ, but the variants was significantly higher in +41170 G homozygotes than in +41170 C homozygotes (P= 0.002). Intron based PCR was was used to amplify transcripts of PBMC pre-mRNA in which intron-5 had been removed (mRNA) while another set of primers was used to amplify intron 5-containing pre-transcripts (pre-mRNA). Knock down analysis of RAB1A Transcripts level in hPBMC. Thromboxane B2 were increased in the siRNA treated when compared those of scramble and control. After knock down analysis, the levels of thromboxane B2 were significantly decreased in PBMC culture supernatant .

Conclusions:

The rare allele of +41170 C>G may play a protective role against aspirin hypersensitivity via a lower catalytic activity of the RAB1A gene attributed to the increase of a non-functioning  variants  of RAB1A.