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Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization

Circulating molecules that are released into the circulation in response to specific stimuli are considered potential biomarkers for physiological or pathological processes. Their effective usefulness as biomarkers resides in their stability and high availability in all the biological fluids, combin...

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Autores principales: Faraldi, Martina, Gomarasca, Marta, Banfi, Giuseppe, Lombardi, Giovanni
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112021/
https://www.ncbi.nlm.nih.gov/pubmed/30342709
http://dx.doi.org/10.1016/bs.acc.2018.07.003
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author Faraldi, Martina
Gomarasca, Marta
Banfi, Giuseppe
Lombardi, Giovanni
author_facet Faraldi, Martina
Gomarasca, Marta
Banfi, Giuseppe
Lombardi, Giovanni
author_sort Faraldi, Martina
collection PubMed
description Circulating molecules that are released into the circulation in response to specific stimuli are considered potential biomarkers for physiological or pathological processes. Their effective usefulness as biomarkers resides in their stability and high availability in all the biological fluids, combined with the limited invasiveness of intervention. Among the circulating molecules, miRNAs represent a novel class of biomarkers as they possess all the required characteristics such as sensitivity, predictivity, specificity, robustness, translatability, and noninvasiveness. miRNAs are small non-coding RNAs, that act as inhibitors of protein translation, and intervene in the complex network of the post-transcriptional mechanisms finely regulating gene expression. The emerging role of miRNAs as potential biomarkers for clinical applications (e.g., cancer and cardiovascular diseases diagnosis and prediction, musculoskeletal disease diagnosis and bone fracture risk prediction), however, requires the standardization of miRNA processing, from sample collection and sample storage, to RNA isolation, RNA reverse-transcription, and data analyses. Normalization is one of the most controversial issues related to quantitative Real-Time PCR data analysis since no universally accepted normalization strategies and reference genes exist, even more importantly, for circulating miRNA quantification. As it is widely demonstrated that the choice of different normalization strategies influences the results of gene expression analysis, it is important to select the most appropriate normalizers for each experimental set. This review discloses on the different strategies adopted in RT-qPCR miRNA normalization and the concerning issues to highlight on the need of a universally accepted methodology to make comparable the results produced by different studies.
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spelling pubmed-71120212020-04-02 Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization Faraldi, Martina Gomarasca, Marta Banfi, Giuseppe Lombardi, Giovanni Adv Clin Chem Article Circulating molecules that are released into the circulation in response to specific stimuli are considered potential biomarkers for physiological or pathological processes. Their effective usefulness as biomarkers resides in their stability and high availability in all the biological fluids, combined with the limited invasiveness of intervention. Among the circulating molecules, miRNAs represent a novel class of biomarkers as they possess all the required characteristics such as sensitivity, predictivity, specificity, robustness, translatability, and noninvasiveness. miRNAs are small non-coding RNAs, that act as inhibitors of protein translation, and intervene in the complex network of the post-transcriptional mechanisms finely regulating gene expression. The emerging role of miRNAs as potential biomarkers for clinical applications (e.g., cancer and cardiovascular diseases diagnosis and prediction, musculoskeletal disease diagnosis and bone fracture risk prediction), however, requires the standardization of miRNA processing, from sample collection and sample storage, to RNA isolation, RNA reverse-transcription, and data analyses. Normalization is one of the most controversial issues related to quantitative Real-Time PCR data analysis since no universally accepted normalization strategies and reference genes exist, even more importantly, for circulating miRNA quantification. As it is widely demonstrated that the choice of different normalization strategies influences the results of gene expression analysis, it is important to select the most appropriate normalizers for each experimental set. This review discloses on the different strategies adopted in RT-qPCR miRNA normalization and the concerning issues to highlight on the need of a universally accepted methodology to make comparable the results produced by different studies. Elsevier Inc. 2018 2018-08-23 /pmc/articles/PMC7112021/ /pubmed/30342709 http://dx.doi.org/10.1016/bs.acc.2018.07.003 Text en Copyright © 2018 Elsevier Inc. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Faraldi, Martina
Gomarasca, Marta
Banfi, Giuseppe
Lombardi, Giovanni
Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization
title Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization
title_full Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization
title_fullStr Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization
title_full_unstemmed Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization
title_short Free Circulating miRNAs Measurement in Clinical Settings: The Still Unsolved Issue of the Normalization
title_sort free circulating mirnas measurement in clinical settings: the still unsolved issue of the normalization
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112021/
https://www.ncbi.nlm.nih.gov/pubmed/30342709
http://dx.doi.org/10.1016/bs.acc.2018.07.003
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