Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease

Background. MicroRNAs (miRNAs) are noncoding RNAs that are highly relevant as disease biomarkers. Several studies that explored the role of miRNAs in Alzheimer’s disease (AD) demonstrated their usefulness in clinical identification. Nevertheless, miRNAs that may act as endogenous controls (ECs) have...

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Autores: Dakterzada, Farida, Targa, Adriano, Benítez, Iván, Romero El Khayat, Leila, de Gonzalo Calvo, David, Torres, Gerard, Moncusí Moix, Anna, Huerto Vilas, Raquel, Sánchez de la Torre, Manuel, Barbé Illa, Ferran, Piñol Ripoll, Gerard
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10459.1/83231
Acceso en línea:https://doi.org/10.1186/s13195-020-00735-x
http://hdl.handle.net/10459.1/83231
Access Level:acceso abierto
Palabra clave:Alzheimer’s disease
Biomarkers
Mild cognitive impairment
miRNAs
Normalization
qPCR
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spelling Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's diseaseDakterzada, FaridaTarga, AdrianoBenítez, IvánRomero El Khayat, Leilade Gonzalo Calvo, DavidTorres, GerardMoncusí Moix, AnnaHuerto Vilas, RaquelSánchez de la Torre, ManuelBarbé Illa, FerranPiñol Ripoll, GerardAlzheimer’s diseaseBiomarkersMild cognitive impairmentmiRNAsNormalizationqPCRBackground. MicroRNAs (miRNAs) are noncoding RNAs that are highly relevant as disease biomarkers. Several studies that explored the role of miRNAs in Alzheimer’s disease (AD) demonstrated their usefulness in clinical identification. Nevertheless, miRNAs that may act as endogenous controls (ECs) have not yet been established. The identification of ECs would contribute to the standardization of these biomarkers in AD. The objective of the study was to identify miRNAs that can be used as ECs in AD. Methods. We evaluated 145 patients divided into two different cohorts. One was a discovery cohort of 19 women diagnosed with mild to moderate AD (Mini-Mental State Examination (MMSE) score ≥ 20) and with confirmed pathologic levels of Aβ42 in CSF. The stability assessment cohort consisted of 126 individuals: 24 subjects without AD or any kind of dementia and negative for all core CSF biomarkers of AD, 25 subjects with MCI and negative for CSF biomarkers (MCI −), 22 subjects with MCI and positive for CSF biomarkers (MCI +), and 55 subjects with AD and positive for CSF biomarkers. In the discovery cohort, a profile of 384 miRNAs was determined in the plasma by TaqMan low-density array. The best EC candidates were identified by mean-centering and concordance correlation restricted normalization methods. The stability of the EC candidates was assessed using the GeNorm, BestKeeper, and NormFinder algorithms. Results. Nine miRNAs (hsa-miR-324-5p, hsa-miR-22-5p, hsa-miR-103a-2-5p, hsa-miR-362-5p, hsa-miR-425-3p, hsa-miR-423-5p, hsa-let-7i-3p, hsa-miR-532-5p, and hsa-miR-1301-3p) were identified as EC candidates in the discovery cohort. The validation results indicated that hsa-miR-103a-2-5p was the best EC, followed by hsa-miR-22-5p, hsa-miR-1301-3p, and hsa-miR-425-3p, which had similar stability values in all three algorithms. Conclusions. We identified a profile of four miRNAs as potential plasma ECs to be used for normalization of miRNA expression data in studies of subjects with cognitive impairment.Generalitat of Catalonia, Department of Health (PERIS 2019 SLT008/18/00050), and “Fundació La Marató TV3” (464/C/2014) to GPR. This study was cofinanced by FEDER funds from the European Union (“A way to build Europe”). IRBLleida is a CERCA Programme/Generalitat of Catalonia. FD was supported by the Agency for Management of University and Research Grants (FI_B100153). David de Gonzalo Calvo acknowledges receiving financial support from Instituto de Salud Carlos III (ISCIII); Miguel Servet 2020: CP20/00041, co-funded by the European Social Fund (ESF)/“Investing in your future”.BCM202220222020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://doi.org/10.1186/s13195-020-00735-xhttp://hdl.handle.net/10459.1/83231http://hdl.handle.net/10459.1/83231reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.1186/s13195-020-00735-xAlzheimer's Research & Therapy, 2020, vol.12cc-by (c) Authors, 2020info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/4.0/oai:recercat.cat:10459.1/832312026-05-29T05:05:01Z
dc.title.none.fl_str_mv Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
title Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
spellingShingle Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
Dakterzada, Farida
Alzheimer’s disease
Biomarkers
Mild cognitive impairment
miRNAs
Normalization
qPCR
title_short Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
title_full Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
title_fullStr Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
title_full_unstemmed Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
title_sort Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease
dc.creator.none.fl_str_mv Dakterzada, Farida
Targa, Adriano
Benítez, Iván
Romero El Khayat, Leila
de Gonzalo Calvo, David
Torres, Gerard
Moncusí Moix, Anna
Huerto Vilas, Raquel
Sánchez de la Torre, Manuel
Barbé Illa, Ferran
Piñol Ripoll, Gerard
author Dakterzada, Farida
author_facet Dakterzada, Farida
Targa, Adriano
Benítez, Iván
Romero El Khayat, Leila
de Gonzalo Calvo, David
Torres, Gerard
Moncusí Moix, Anna
Huerto Vilas, Raquel
Sánchez de la Torre, Manuel
Barbé Illa, Ferran
Piñol Ripoll, Gerard
author_role author
author2 Targa, Adriano
Benítez, Iván
Romero El Khayat, Leila
de Gonzalo Calvo, David
Torres, Gerard
Moncusí Moix, Anna
Huerto Vilas, Raquel
Sánchez de la Torre, Manuel
Barbé Illa, Ferran
Piñol Ripoll, Gerard
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Alzheimer’s disease
Biomarkers
Mild cognitive impairment
miRNAs
Normalization
qPCR
topic Alzheimer’s disease
Biomarkers
Mild cognitive impairment
miRNAs
Normalization
qPCR
description Background. MicroRNAs (miRNAs) are noncoding RNAs that are highly relevant as disease biomarkers. Several studies that explored the role of miRNAs in Alzheimer’s disease (AD) demonstrated their usefulness in clinical identification. Nevertheless, miRNAs that may act as endogenous controls (ECs) have not yet been established. The identification of ECs would contribute to the standardization of these biomarkers in AD. The objective of the study was to identify miRNAs that can be used as ECs in AD. Methods. We evaluated 145 patients divided into two different cohorts. One was a discovery cohort of 19 women diagnosed with mild to moderate AD (Mini-Mental State Examination (MMSE) score ≥ 20) and with confirmed pathologic levels of Aβ42 in CSF. The stability assessment cohort consisted of 126 individuals: 24 subjects without AD or any kind of dementia and negative for all core CSF biomarkers of AD, 25 subjects with MCI and negative for CSF biomarkers (MCI −), 22 subjects with MCI and positive for CSF biomarkers (MCI +), and 55 subjects with AD and positive for CSF biomarkers. In the discovery cohort, a profile of 384 miRNAs was determined in the plasma by TaqMan low-density array. The best EC candidates were identified by mean-centering and concordance correlation restricted normalization methods. The stability of the EC candidates was assessed using the GeNorm, BestKeeper, and NormFinder algorithms. Results. Nine miRNAs (hsa-miR-324-5p, hsa-miR-22-5p, hsa-miR-103a-2-5p, hsa-miR-362-5p, hsa-miR-425-3p, hsa-miR-423-5p, hsa-let-7i-3p, hsa-miR-532-5p, and hsa-miR-1301-3p) were identified as EC candidates in the discovery cohort. The validation results indicated that hsa-miR-103a-2-5p was the best EC, followed by hsa-miR-22-5p, hsa-miR-1301-3p, and hsa-miR-425-3p, which had similar stability values in all three algorithms. Conclusions. We identified a profile of four miRNAs as potential plasma ECs to be used for normalization of miRNA expression data in studies of subjects with cognitive impairment.
publishDate 2020
dc.date.none.fl_str_mv 2020
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://doi.org/10.1186/s13195-020-00735-x
http://hdl.handle.net/10459.1/83231
http://hdl.handle.net/10459.1/83231
url https://doi.org/10.1186/s13195-020-00735-x
http://hdl.handle.net/10459.1/83231
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1186/s13195-020-00735-x
Alzheimer's Research & Therapy, 2020, vol.12
dc.rights.none.fl_str_mv cc-by (c) Authors, 2020
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/4.0/
rights_invalid_str_mv cc-by (c) Authors, 2020
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BCM
publisher.none.fl_str_mv BCM
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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