SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume.
Genome-wide association studies have identified a genetic variant at 3p14.3 (SNP rs1354034) that strongly associates with platelet number and mean platelet volume in humans. While originally proposed to be intronic, analysis of mRNA expression in primary human hematopoietic subpopulations reveals th...
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Public Library of Science (PLoS)
2017-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0178095&type=printable |
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| author | Siying Zou Alexandra M Teixeira Myrto Kostadima William J Astle Aparna Radhakrishnan Lukas Mikolaj Simon Lucy Truman Jennifer S Fang John Hwa Ping-Xia Zhang Pim van der Harst Paul F Bray Willem H Ouwehand Mattia Frontini Diane S Krause |
| author_facet | Siying Zou Alexandra M Teixeira Myrto Kostadima William J Astle Aparna Radhakrishnan Lukas Mikolaj Simon Lucy Truman Jennifer S Fang John Hwa Ping-Xia Zhang Pim van der Harst Paul F Bray Willem H Ouwehand Mattia Frontini Diane S Krause |
| author_sort | Siying Zou |
| collection | DOAJ |
| description | Genome-wide association studies have identified a genetic variant at 3p14.3 (SNP rs1354034) that strongly associates with platelet number and mean platelet volume in humans. While originally proposed to be intronic, analysis of mRNA expression in primary human hematopoietic subpopulations reveals that this SNP is located directly upstream of the predominantly expressed ARHGEF3 isoform in megakaryocytes (MK). We found that ARHGEF3, which encodes a Rho guanine exchange factor, is dramatically upregulated during both human and murine MK maturation. We show that the SNP (rs1354034) is located in a DNase I hypersensitive region in human MKs and is an expression quantitative locus (eQTL) associated with ARHGEF3 expression level in human platelets, suggesting that it may be the causal SNP that accounts for the variations observed in human platelet traits and ARHGEF3 expression. In vitro human platelet activation assays revealed that rs1354034 is highly correlated with human platelet activation by ADP. In order to test whether ARHGEF3 plays a role in MK development and/or platelet function, we developed an Arhgef3 KO/LacZ reporter mouse model. Reflecting changes in gene expression, LacZ expression increases during MK maturation in these mice. Although Arhgef3 KO mice have significantly larger platelets, loss of Arhgef3 does not affect baseline MK or platelets nor does it affect platelet function or platelet recovery in response to antibody-mediated platelet depletion compared to littermate controls. In summary, our data suggest that modulation of ARHGEF3 gene expression in humans with a promoter-localized SNP plays a role in human MKs and human platelet function-a finding resulting from the biological follow-up of human genetic studies. Arhgef3 KO mice partially recapitulate the human phenotype. |
| format | Article |
| id | doaj-art-bb909eba264445a1a24ac7e5e257a0c7 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-bb909eba264445a1a24ac7e5e257a0c72025-08-20T03:24:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01125e017809510.1371/journal.pone.0178095SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume.Siying ZouAlexandra M TeixeiraMyrto KostadimaWilliam J AstleAparna RadhakrishnanLukas Mikolaj SimonLucy TrumanJennifer S FangJohn HwaPing-Xia ZhangPim van der HarstPaul F BrayWillem H OuwehandMattia FrontiniDiane S KrauseGenome-wide association studies have identified a genetic variant at 3p14.3 (SNP rs1354034) that strongly associates with platelet number and mean platelet volume in humans. While originally proposed to be intronic, analysis of mRNA expression in primary human hematopoietic subpopulations reveals that this SNP is located directly upstream of the predominantly expressed ARHGEF3 isoform in megakaryocytes (MK). We found that ARHGEF3, which encodes a Rho guanine exchange factor, is dramatically upregulated during both human and murine MK maturation. We show that the SNP (rs1354034) is located in a DNase I hypersensitive region in human MKs and is an expression quantitative locus (eQTL) associated with ARHGEF3 expression level in human platelets, suggesting that it may be the causal SNP that accounts for the variations observed in human platelet traits and ARHGEF3 expression. In vitro human platelet activation assays revealed that rs1354034 is highly correlated with human platelet activation by ADP. In order to test whether ARHGEF3 plays a role in MK development and/or platelet function, we developed an Arhgef3 KO/LacZ reporter mouse model. Reflecting changes in gene expression, LacZ expression increases during MK maturation in these mice. Although Arhgef3 KO mice have significantly larger platelets, loss of Arhgef3 does not affect baseline MK or platelets nor does it affect platelet function or platelet recovery in response to antibody-mediated platelet depletion compared to littermate controls. In summary, our data suggest that modulation of ARHGEF3 gene expression in humans with a promoter-localized SNP plays a role in human MKs and human platelet function-a finding resulting from the biological follow-up of human genetic studies. Arhgef3 KO mice partially recapitulate the human phenotype.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0178095&type=printable |
| spellingShingle | Siying Zou Alexandra M Teixeira Myrto Kostadima William J Astle Aparna Radhakrishnan Lukas Mikolaj Simon Lucy Truman Jennifer S Fang John Hwa Ping-Xia Zhang Pim van der Harst Paul F Bray Willem H Ouwehand Mattia Frontini Diane S Krause SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. PLoS ONE |
| title | SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. |
| title_full | SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. |
| title_fullStr | SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. |
| title_full_unstemmed | SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. |
| title_short | SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. |
| title_sort | snp in human arhgef3 promoter is associated with dnase hypersensitivity transcript level and platelet function and arhgef3 ko mice have increased mean platelet volume |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0178095&type=printable |
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