Identification of variation in the platelet transcriptome associated with glycoprotein 6 haplotype

Platelets. 2008 Jun;19(4):258-67. doi: 10.1080/09537100801947434.

Abstract

Platelet Glycoprotein VI (GPVI) is the activatory collagen signalling receptor that transmits an outside-in signal via the FcR gamma-chain. In Caucasians two GP6 haplotypes have been identified which encode GPVI isoforms that differ by five amino-acids. The minor haplotype is associated with a modest but statistically significant reduction in GPVI abundance and reduced downstream signalling events. As GPVI is also expressed on megakaryocytes, different GPVI isoforms may imprint on the platelet transcriptome. We investigated the association of GP6 haplotype with transcription by comparing the transcriptomes of platelets from individuals homozygous for the major ('a') and minor ('b') haplotypes to identify differentially expressed (DE) transcripts. Platelet RNA was isolated from apheresis concentrates from 16 'aa' donors and eight 'bb' donors. mRNA was amplified using a template-switching PCR based protocol and fluorescently labelled. Samples were randomly paired both within and between haplotypes and compared on a cDNA microarray. No consistently DE transcripts were identified within the 'aa' haplotype but 52 significantly DE transcripts were observed between haplotypes. Generally the fold differences were low (two to four-fold) but were confirmed by qRT-PCR for selected transcripts (TUBB1, P = 0.0004; VWF, P = 0.0126). The results of this study indicate that there are subtle differences between the platelet transcriptomes of individuals who differ by GP6 haplotype. The identification of DE genes may identify critical pathways and nodes not previously known to be involved in platelet development and function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Platelets / chemistry*
  • Gene Expression Profiling*
  • Genetic Variation*
  • Haplotypes*
  • Humans
  • Megakaryocytes
  • Platelet Membrane Glycoproteins / genetics*
  • RNA, Messenger / analysis*

Substances

  • Platelet Membrane Glycoproteins
  • RNA, Messenger
  • platelet membrane glycoprotein VI