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Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease.
Alzheimers Res Ther. 2020 12 05; 12(1):163.AR

Abstract

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.

Authors+Show Affiliations

Unitat Trastorns Cognitius, Clinical Neuroscience Research, Hospital Universitari de Santa Maria, IRBLleida, Lleida, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain. Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Madrid, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain. Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Madrid, Spain.Unitat Trastorns Cognitius, Clinical Neuroscience Research, Hospital Universitari de Santa Maria, IRBLleida, Lleida, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain. Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Madrid, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain. Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Madrid, Spain.Unitat Trastorns Cognitius, Clinical Neuroscience Research, Hospital Universitari de Santa Maria, IRBLleida, Lleida, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain. Group of Precision Medicine in Chronic Diseases, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain.Translational Research in Respiratory Medicine, Hospital Universitari Arnau de Vilanova-Santa Maria, IRBLleida, Lleida, Spain. Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Madrid, Spain.Unitat Trastorns Cognitius, Clinical Neuroscience Research, Hospital Universitari de Santa Maria, IRBLleida, Lleida, Spain. gerard_437302@hotmail.com.

Pub Type(s)

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

Language

eng

PubMed ID

33278902

Citation

Dakterzada, F, et al. "Identification and Validation of Endogenous Control miRNAs in Plasma Samples for Normalization of qPCR Data for Alzheimer's Disease." Alzheimer's Research & Therapy, vol. 12, no. 1, 2020, p. 163.
Dakterzada F, Targa A, Benítez ID, et al. Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease. Alzheimers Res Ther. 2020;12(1):163.
Dakterzada, F., Targa, A., Benítez, I. D., Romero-ElKhayat, L., de Gonzalo-Calvo, D., Torres, G., Moncusí-Moix, A., Huerto, R., Sánchez-de-la-Torre, M., Barbé, F., & Piñol-Ripoll, G. (2020). Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease. Alzheimer's Research & Therapy, 12(1), 163. https://doi.org/10.1186/s13195-020-00735-x
Dakterzada F, et al. Identification and Validation of Endogenous Control miRNAs in Plasma Samples for Normalization of qPCR Data for Alzheimer's Disease. Alzheimers Res Ther. 2020 12 5;12(1):163. PubMed PMID: 33278902.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer's disease. AU - Dakterzada,F, AU - Targa,A, AU - Benítez,I D, AU - Romero-ElKhayat,L, AU - de Gonzalo-Calvo,D, AU - Torres,G, AU - Moncusí-Moix,A, AU - Huerto,R, AU - Sánchez-de-la-Torre,M, AU - Barbé,F, AU - Piñol-Ripoll,G, Y1 - 2020/12/05/ PY - 2020/07/29/received PY - 2020/11/26/accepted PY - 2020/12/6/entrez PY - 2020/12/7/pubmed PY - 2021/6/25/medline KW - Alzheimer’s disease KW - Biomarkers KW - Mild cognitive impairment KW - Normalization KW - miRNAs KW - qPCR SP - 163 EP - 163 JF - Alzheimer's research & therapy JO - Alzheimers Res Ther VL - 12 IS - 1 N2 - 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. SN - 1758-9193 UR - https://www.unboundmedicine.com/medline/citation/33278902/Identification_and_validation_of_endogenous_control_miRNAs_in_plasma_samples_for_normalization_of_qPCR_data_for_Alzheimer's_disease_ DB - PRIME DP - Unbound Medicine ER -