Platelet Transcriptome and Cardiovascular Disease

Senzel, Lisa; Gnatenko, Dmitri V.; Bahou, Wadie F.

doi:10.2217/14796678.3.4.391

Cited by 2 publications

(2 citation statements)

References 42 publications

(53 reference statements)

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“…Differentially-expressed mRNAs can potentially be used as biomarkers for diagnostics and prognostics of platelet-related diseases. Comparison of platelet transcriptomes from healthy individuals and patient cohorts with distinct haematologic and cardiovascular diseases reveals significant differences in the spectrum of platelet-expressed mRNAs (6,7,20). Nonetheless, traditional technological platforms for platelet transcript profiling (microarray, quantitative real-time RT-PCR [Q-PCR], serial analysis of gene expression [SAGE], large-scale sequencing, etc.)…”

Section: Introductionmentioning

confidence: 99%

Platelet genetic biomarker quantification: Comparison of fluorescent microspheres and PCR platforms

Huang¹,

Zhu²,

Dhundale³

et al. 2013

Thromb Haemost

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The platelet transcriptome has been extensively characterised using distinct genetic profiling platforms, with evolving evidence for differential expression patterns between healthy individuals and subject cohorts with various haematologic and cardiovascular disorders. Traditional technological platforms for platelet genetic biomarker quantification have limited applicability for clinical molecular diagnostics due to inherent complexities related to RNA isolation and analysis. We have previously established the feasibility of fluorescent microspheres as a simple and reproducible strategy for simultaneous quantification of platelet mRNAs from small volume of blood using intact platelets. We now extend these observations by formally comparing in a 50-member normal cohort the cross-platform behaviour of fluorescent microspheres to the currently accepted Q-PCR standard, using a clinically relevant 15-biomarker gene subset able to discriminate among normal and thrombocytosis cohorts. When compared to Q-PCR, genetic biomarker quantification using fluorescent microspheres demonstrated lower coefficients of variation for low-abundant transcripts, better linearity in serially diluted samples, and good overall between-platform consistency via the geometric mean regression. Neither platform demonstrated age or gender effects for any of the 15 biomarkers studied. Binding site saturation for highly abundant transcripts using fluorescent microspheres can be readily eliminated using an optimal platelet number corresponding to 0.3 ml of peripheral blood, additionally applicable to thrombocytopenic cohorts. These data provide a detailed cross-platform analysis using a relevant biomarker subset, further highlighting the applicability of fluorescent microspheres as potentially superior to Q-PCR for platelet mRNA diagnostics.

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Section: Introductionmentioning

confidence: 99%

Platelet genetic biomarker quantification: Comparison of fluorescent microspheres and PCR platforms

Huang¹,

Zhu²,

Dhundale³

et al. 2013

Thromb Haemost

View full text Add to dashboard Cite

show abstract

“…Current methodsofplatelettranscriptprofiling (microarray, serialanalysis of gene expression [SAGE],and quantitative realtime reverse-transcription PCR [Q-PCR])represent well-establishedtechniquesthat arepoorlyadaptedtoclinical laboratories due to the complexity of RNAi solation, experimental design, and subsequent data analysis (4,(12)(13)(14). Towards adapting transcript profiling technologiest oc linical use, we appliedar ecently-developedplatformfor rapid and efficient platelet mRNA profiling.…”

Section: Introductionmentioning

confidence: 99%

Multiplexed genetic profiling of human blood platelets using fluorescent microspheres

Gnatenko

Zhu²,

Bahou³

2008

Thromb Haemost

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Human platelets have unique and reproducible mRNA profiles, with evidence for distinct profiles in haematopoietic stem cell disorders associated with thrombocytosis. Platelet transcript profiling is traditionally studied by microarray analysis, quantitative reverse transcription-PCR or serial analysis of gene expression, techniques that are labor- and technically-intensive. We have now applied a novel multiplex-based platform for quantitative transcript profiling of human platelets. Simultaneous quantification of 17 platelet transcripts was assayed using intact platelet-rich plasma or gel-filtered platelets lysed in vitro. Accurate and reproducible profiles could be obtained from as few as 5 x 10(7) platelets (a platelet mass corresponding to approximately 100 microl of whole blood), even for the low-abundant platelet transcripts. Correlation coefficients of this 17-member gene set to platelet Affymetrix microarrays were excellent (r(2) = 0.949, p < 1 x 10(-10)), with no correlation to in kind-derived leukocyte profiles, highlighting the cell-specificity of the platform. These data demonstrate that transcript multiplexing using fluorescent microspheres can be adapted for rapid molecular profiling using intact platelets (bypassing the need for RNA isolation methods), with potential applicability irrespective of baseline platelet counts.

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Platelet Transcriptome and Cardiovascular Disease

Cited by 2 publications

References 42 publications

Platelet genetic biomarker quantification: Comparison of fluorescent microspheres and PCR platforms

Platelet genetic biomarker quantification: Comparison of fluorescent microspheres and PCR platforms

Multiplexed genetic profiling of human blood platelets using fluorescent microspheres

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