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Bringing to Light the Importance of the miRNA Methylome in Colorectal Cancer Prognosis Through Electrochemical Bioplatforms

This work reports the first electrochemical bioplatforms developed for the determination of the total contents of either target miRNA or methylated target miRNA. The bioplatforms are based on the hybridization of the target miRNA with a synthetic biotinylated DNA probe, the capture of the formed DNA...

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Published in:Analytical chemistry (Washington) 2024-03, Vol.96 (11), p.4580-4588
Main Authors: Povedano, Eloy, Ruiz-Valdepeñas Montiel, Víctor, Sebuyoya, Ravery, Torrente-Rodríguez, Rebeca M., Garranzo-Asensio, Maria, Montero-Calle, Ana, Pingarrón, José M., Barderas, Rodrigo, Bartosik, Martin, Campuzano, Susana
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creator Povedano, Eloy
Ruiz-Valdepeñas Montiel, Víctor
Sebuyoya, Ravery
Torrente-Rodríguez, Rebeca M.
Garranzo-Asensio, Maria
Montero-Calle, Ana
Pingarrón, José M.
Barderas, Rodrigo
Bartosik, Martin
Campuzano, Susana
description This work reports the first electrochemical bioplatforms developed for the determination of the total contents of either target miRNA or methylated target miRNA. The bioplatforms are based on the hybridization of the target miRNA with a synthetic biotinylated DNA probe, the capture of the formed DNA/miRNA heterohybrids on the surface of magnetic microcarriers, and their recognition with an antibody selective to these heterohybrids or to the N 6-methyladenosine (m6A) epimark. The determination of the total or methylated target miRNA was accomplished by labeling such secondary antibodies with the horseradish peroxidase (HRP) enzyme. In both cases, amperometric transduction was performed on the surface of disposable electrodes after capturing the resulting HRP-tagged magnetic bioconjugates. Because of their increasing relevance in colorectal cancer (CRC) diagnosis and prognosis, miRNA let-7a and m6A methylation were selected. The proposed electrochemical bioplatforms showed attractive analytical and operational characteristics for the determination of the total and m6A-methylated target miRNA in less than 75 min. These bioplatforms, innovative in design and application, were applied to the analysis of total RNA samples extracted from cultured cancer cells with different metastatic profiles and from paired healthy and tumor tissues of patients diagnosed with CRC at different stages. The obtained results demonstrated, for the first time using electrochemical platforms, the potential of interrogating the target miRNA methylation level to discriminate the metastatic capacities of cancer cells and to identify tumor tissues and, in a pioneering way, the potential of the m6A methylation in miRNA let-7a to serve as a prognostic biomarker for CRC.
doi_str_mv 10.1021/acs.analchem.3c05474
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The bioplatforms are based on the hybridization of the target miRNA with a synthetic biotinylated DNA probe, the capture of the formed DNA/miRNA heterohybrids on the surface of magnetic microcarriers, and their recognition with an antibody selective to these heterohybrids or to the N 6-methyladenosine (m6A) epimark. The determination of the total or methylated target miRNA was accomplished by labeling such secondary antibodies with the horseradish peroxidase (HRP) enzyme. In both cases, amperometric transduction was performed on the surface of disposable electrodes after capturing the resulting HRP-tagged magnetic bioconjugates. Because of their increasing relevance in colorectal cancer (CRC) diagnosis and prognosis, miRNA let-7a and m6A methylation were selected. The proposed electrochemical bioplatforms showed attractive analytical and operational characteristics for the determination of the total and m6A-methylated target miRNA in less than 75 min. 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subjects Antibodies
Antibodies - genetics
Biomarkers
Biosensing Techniques - methods
Cancer
Colorectal cancer
Colorectal carcinoma
Colorectal Neoplasms - diagnosis
Colorectal Neoplasms - genetics
Deoxyribonucleic acid
DNA
Electrical measurement
Electrochemistry
Epigenome
Horseradish peroxidase
Humans
Hybridization
Medical prognosis
Metastases
Metastasis
Methylation
MicroRNAs - analysis
MicroRNAs - genetics
miRNA
N6-methyladenosine
Neoplasms
Nucleic Acid Hybridization - methods
Peroxidase
Prognosis
Transduction
Tumors
title Bringing to Light the Importance of the miRNA Methylome in Colorectal Cancer Prognosis Through Electrochemical Bioplatforms
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