Novel loci involved in platelet function and platelet count identified by a genome-wide study performed in children

Abstract

Background: Genome-wide association studies are currently identifying new loci with potential roles in thrombosis and hemostasis: these loci include novel polymorphisms associated with platelet function traits and count. However, no genome-wide study performed on children has been reported to date, in spite of the potential that these subjects have in genetic studies, when compared to adults, given the minimal degree of confounders, i.e., acquired and environmental factors, such as smoking, physical activity, diet, and drug or hormone intake, which are particularly important in platelet function.

Design and methods: To identify new genetic variants involved in platelet reactivity and count, we performed a genome-wide association study on 75 children (8.5±1.8 years) using the Illumina Sentrix Human CNV370-Quad BeadChip containing 320,610 single nucleotide polymorphisms. Functional analyses included assessment of platelet aggregation and granule secretion triggered by different agonists (arachidonic acid, collagen, epinephrine, ADP), as well as platelet count. Associations were selected based on statistical significance and physiological relevance for a subsequent replication study in a similar sample of 286 children.

Results: We confirmed previously established associations with plasma levels of factors XII, VII and VIII as well as associations with platelet responses to ADP. Additionally, we identified 82 associations with platelet reactivity and count with a P value less than 10(-5). From the associations selected for further replication, we validated two single nucleotide polymorphisms with mildly increased platelet reactivity (rs4366150 and rs1787566) on the LPAR1 and MYO5B genes, encoding lisophosphatidic acid receptor-1 and myosin VB, respectively; and rs1937970, located on the NRG3 gene coding neuroregulin-3, associated with platelet count.

Conclusions: Our genome-wide association study performed in children, followed by a validation analysis, led us to the identification of new genes potentially relevant in platelet function and biogenesis.

Figure 1.

Effect of rs4366150 (LPAR1) on platelet aggregation and secretion assays. (A) Data from the INGENIAHS. In all cases P value <0.01. (B) Data from the replication population. *indicates a P value <0.05 when AA controls were compared to AG+GG individuals in the replication study. Results represent mean ± SD.

Figure 2.

Effect of rs1787566 (MYO5B) on platelet aggregation and secretion. (A) Data from the INGENIAHS. In all cases P value <0.01. (B) Results obtained in the validation population. *indicates a P value <0.05 when AA controls were compared to AG+GG individuals in the replication study. Results represent mean ± SD.

Figure 3.

Effect of rs1937970 (NRG3) on platelet count. (A) Data from the INGENIAHS. P value = 3.55 x 10⁻⁵. (B) Results obtained in the validation population. *indicates a P value <0.05 when AA controls were compared to AG+GG individuals in the replication study. Results represent mean ± SD.