Circular RNA Sequencing of Maternal Platelets: A Novel Tool for the Identification of Pregnancy-Specific Biomarkers.


Journal

Clinical chemistry
ISSN: 1530-8561
Titre abrégé: Clin Chem
Pays: England
ID NLM: 9421549

Informations de publication

Date de publication:
01 03 2021
Historique:
received: 26 05 2020
accepted: 29 09 2020
pubmed: 2 12 2020
medline: 24 9 2021
entrez: 1 12 2020
Statut: ppublish

Résumé

In the first trimester of pregnancy, the maternal platelet is directly involved in a positive feedback mechanism that facilitates invasion of the extravillous trophoblast into the maternal spiral arteries. Dysfunctional trophoblast invasion with defective deep placentation is primordial in the etiology of the "great obstetrical syndromes." In this proof-of-concept study, using transcriptome analysis of circular RNA (circRNA) following RNA sequencing of maternal platelets, we tested whether pregnancy-specific circRNA markers could be identified in the first trimester of normal pregnancies. Differential transcript expression analysis of circRNAs, as predicted by Accurate CircRNA Finder Suite, CircRNA Identifier (version 2), and Known and Novel Isoform Explorer, was done using thromboSeq.R with variation of multiple settings. Test performance was checked for (a) de novo circRNA identification using the novel platelet-specific Plt-circR4 as a positive control, (b) complete segregation of groups (pregnant vs nonpregnant) after heat map-dendrogram clustering, (c) identification of pregnancy-specific circRNA markers at a false discovery rate (FDR) <0.05, and (d) confirmation of differentially expressed circRNA markers with an FDR <0.05 by an independent method, reverse transcription-quantitative PCR. Of the differentially expressed circRNAs with P values <0.05, 41 circRNAs were upregulated (logFC >2), and 52 circRNAs were downregulated (logFC less than -2) in first-trimester platelet RNA. Of these, nuclear receptor-interacting protein 1 circRNA covering exons 2 and 3 of the 5'-untranslated region was pregnancy specific with upregulation in first-trimester maternal platelets compared to nonpregnant controls. CircRNA sequencing of first-trimester maternal platelets permits the identification of novel pregnancy-specific RNA biomarkers. Future use could include the assessment of maternal and fetal well-being.

Sections du résumé

BACKGROUND
In the first trimester of pregnancy, the maternal platelet is directly involved in a positive feedback mechanism that facilitates invasion of the extravillous trophoblast into the maternal spiral arteries. Dysfunctional trophoblast invasion with defective deep placentation is primordial in the etiology of the "great obstetrical syndromes."
METHODS
In this proof-of-concept study, using transcriptome analysis of circular RNA (circRNA) following RNA sequencing of maternal platelets, we tested whether pregnancy-specific circRNA markers could be identified in the first trimester of normal pregnancies. Differential transcript expression analysis of circRNAs, as predicted by Accurate CircRNA Finder Suite, CircRNA Identifier (version 2), and Known and Novel Isoform Explorer, was done using thromboSeq.R with variation of multiple settings. Test performance was checked for (a) de novo circRNA identification using the novel platelet-specific Plt-circR4 as a positive control, (b) complete segregation of groups (pregnant vs nonpregnant) after heat map-dendrogram clustering, (c) identification of pregnancy-specific circRNA markers at a false discovery rate (FDR) <0.05, and (d) confirmation of differentially expressed circRNA markers with an FDR <0.05 by an independent method, reverse transcription-quantitative PCR.
RESULTS
Of the differentially expressed circRNAs with P values <0.05, 41 circRNAs were upregulated (logFC >2), and 52 circRNAs were downregulated (logFC less than -2) in first-trimester platelet RNA. Of these, nuclear receptor-interacting protein 1 circRNA covering exons 2 and 3 of the 5'-untranslated region was pregnancy specific with upregulation in first-trimester maternal platelets compared to nonpregnant controls.
CONCLUSION
CircRNA sequencing of first-trimester maternal platelets permits the identification of novel pregnancy-specific RNA biomarkers. Future use could include the assessment of maternal and fetal well-being.

Identifiants

pubmed: 33257975
pii: 6012968
doi: 10.1093/clinchem/hvaa249
doi:

Substances chimiques

Biomarkers 0
RNA, Circular 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

508-517

Informations de copyright

© American Association for Clinical Chemistry 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Auteurs

Cees Oudejans (C)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Vera Manders (V)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.
Reproductive Biology Laboratory, Amsterdam UMC, Academic Medical Center, Amsterdam, the Netherlands.

Allerdien Visser (A)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Remco Keijser (R)

Reproductive Biology Laboratory, Amsterdam UMC, Academic Medical Center, Amsterdam, the Netherlands.

Naomi Min (N)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.
Reproductive Biology Laboratory, Amsterdam UMC, Academic Medical Center, Amsterdam, the Netherlands.

Ankie Poutsma (A)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Joyce Mulders (J)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Tarah van den Berkmortel (T)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Di-Jan Wigman (DJ)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Britt Blanken (B)

Department of Clinical Chemistry, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Aldo Jongejan (A)

Department of Clinical Epidemiology, Biostatistics, and Bioinformatics, Amsterdam UMC, Academic Medical Center, the Netherlands.

Eva Pajkrt (E)

Department of Obstetrics/Gynecology, Amsterdam UMC, Academic Medical Center, Amsterdam, the Netherlands.

Marjon de Boer (M)

Department of Obstetrics/Gynecology, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Erik A Sistermans (EA)

Department of Clinical Genetics, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Daoud Sie (D)

Department of Clinical Genetics, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Myron G Best (MG)

Department of Neurosurgery, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.
Department of Pathology, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.
Brain Tumor Center Amsterdam, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Tom Würdinger (T)

Department of Neurosurgery, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.
Brain Tumor Center Amsterdam, Amsterdam UMC, VU University Medical Center, Amsterdam, the Netherlands.

Gijs Afink (G)

Reproductive Biology Laboratory, Amsterdam UMC, Academic Medical Center, Amsterdam, the Netherlands.

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Classifications MeSH