Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheime
- PDF / 501,138 Bytes
- 8 Pages / 595.276 x 790.866 pts Page_size
- 6 Downloads / 160 Views
(2020) 12:163
RESEARCH
Open Access
Identification and validation of endogenous control miRNAs in plasma samples for normalization of qPCR data for Alzheimer’s disease F. Dakterzada1†, A. Targa2,3†, I. D. Benítez2,3, L. Romero-ElKhayat1, D. de Gonzalo-Calvo2, G. Torres2,3, A. Moncusí-Moix2,3, R. Huerto1, M. Sánchez-de-la-Torre2,4, F. Barbé2,3 and G. Piñol-Ripoll1*
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, hsamiR-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, hsamiR-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. Keywords: Alzheimer’s disease, Biomarkers, Mild cognitive impairment, miRNAs, Normalization, qPCR
* Correspondence: [email protected] † Dakterzada, F. and Targa, A. are co-first authors and contributed equally to this study. 1 Unitat Trastorns Cognitius, Clinical Neuroscience Research, Hospital Universitari de Santa Maria, IRBLleida, Lleida, Spain Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
Data Loading...
