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      From sequencing to validation: NGS-based exploration of plasma miRNA in papillary thyroid carcinoma

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          Abstract

          Objective

          A non-invasive method using plasma microRNAs provides new insights into thyroid cancer diagnosis. The objective of this study was to discover potential circulating biomarkers of papillary thyroid carcinoma (PTC) through the analysis of plasma miRNAs using next-generation sequencing (NGS).

          Methods

          Plasma miRNAs were isolated from peripheral blood samples collected from healthy individuals, patients diagnosed with PTC, and those with benign thyroid nodules. The Illumina NovaSeq 6000 platform was employed to establish the miRNA expression profiles. Candidate miRNAs for diagnostic purposes were identified utilizing the Random Forest (RF) algorithm. The selected miRNAs were subsequently validated in an independent validation set using RT-qPCR.

          Results

          NGS results revealed consistent plasma miRNA expression patterns among healthy individuals and patients with benign thyroid nodules in the discovery set (6 healthy cases, 17 benign cases), while differing significantly from those observed in the PTC group (17 PTC cases). Seven miRNAs exhibiting significant expression differences were identified and utilized to construct an RF classifier. Receiver operating characteristic (ROC) analysis for PTC diagnosis, and the area under the curve (AUC) was 0.978. Subsequent KEGG and GO analyses of the target genes associated with these 7 miRNAs highlighted pathways relevant to tumors and the cell cycle. Independent validation through RT-qPCR in a separate cohort (15 CONTROL, 15 PTC groups) underscored hsa-miR-301a-3p and hsa-miR-195-5p as promising candidates for PTC diagnosis.

          Conclusion

          In conclusion, our study established a seven-miRNA panel in plasma by Random Forest algorithm with significant performance in discriminating PTC from healthy or benign group. hsa-miR-301a-3p, hsa-miR-195-5p in plasma have potential for further study in the diagnosis of PTC in Asian ethnic.

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          Most cited references75

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          Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries

          This article provides an update on the global cancer burden using the GLOBOCAN 2020 estimates of cancer incidence and mortality produced by the International Agency for Research on Cancer. Worldwide, an estimated 19.3 million new cancer cases (18.1 million excluding nonmelanoma skin cancer) and almost 10.0 million cancer deaths (9.9 million excluding nonmelanoma skin cancer) occurred in 2020. Female breast cancer has surpassed lung cancer as the most commonly diagnosed cancer, with an estimated 2.3 million new cases (11.7%), followed by lung (11.4%), colorectal (10.0 %), prostate (7.3%), and stomach (5.6%) cancers. Lung cancer remained the leading cause of cancer death, with an estimated 1.8 million deaths (18%), followed by colorectal (9.4%), liver (8.3%), stomach (7.7%), and female breast (6.9%) cancers. Overall incidence was from 2-fold to 3-fold higher in transitioned versus transitioning countries for both sexes, whereas mortality varied <2-fold for men and little for women. Death rates for female breast and cervical cancers, however, were considerably higher in transitioning versus transitioned countries (15.0 vs 12.8 per 100,000 and 12.4 vs 5.2 per 100,000, respectively). The global cancer burden is expected to be 28.4 million cases in 2040, a 47% rise from 2020, with a larger increase in transitioning (64% to 95%) versus transitioned (32% to 56%) countries due to demographic changes, although this may be further exacerbated by increasing risk factors associated with globalization and a growing economy. Efforts to build a sustainable infrastructure for the dissemination of cancer prevention measures and provision of cancer care in transitioning countries is critical for global cancer control.
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            Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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              edgeR: a Bioconductor package for differential expression analysis of digital gene expression data

              Summary: It is expected that emerging digital gene expression (DGE) technologies will overtake microarray technologies in the near future for many functional genomics applications. One of the fundamental data analysis tasks, especially for gene expression studies, involves determining whether there is evidence that counts for a transcript or exon are significantly different across experimental conditions. edgeR is a Bioconductor software package for examining differential expression of replicated count data. An overdispersed Poisson model is used to account for both biological and technical variability. Empirical Bayes methods are used to moderate the degree of overdispersion across transcripts, improving the reliability of inference. The methodology can be used even with the most minimal levels of replication, provided at least one phenotype or experimental condition is replicated. The software may have other applications beyond sequencing data, such as proteome peptide count data. Availability: The package is freely available under the LGPL licence from the Bioconductor web site (http://bioconductor.org). Contact: mrobinson@wehi.edu.au

                Author and article information

                Contributors
                URI : https://loop.frontiersin.org/people/2701551Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/2725020Role: Role: Role: Role:
                Role: Role:
                URI : https://loop.frontiersin.org/people/1296050Role: Role: Role: Role:
                Journal
                Front Oncol
                Front Oncol
                Front. Oncol.
                Frontiers in Oncology
                Frontiers Media S.A.
                2234-943X
                07 August 2024
                2024
                : 14
                : 1410110
                Affiliations
                [1] 1 Department of Head and Neck Surgery, Fudan University Shanghai Cancer Center, Fudan University , Shanghai, China
                [2] 2 Department of Oncology, Shanghai Medical College, Fudan University , Shanghai, China
                [3] 3 Runan Medical Technology (Suzhou) Co., LTD , Suzhou, China
                [4] 4 Shanghai Runan Medical Technology Co., LTD , Shanghai, China
                Author notes

                Edited by: Zein Mersini Besharat, Sapienza University of Rome, Italy

                Reviewed by: Thomas Kleen, Biolab Services LLC, United States

                Stefania Mardente, Sapienza University of Rome, Italy

                *Correspondence: Yu Wang, neck130@ 123456sina.com ; Tian Liao, liaotian@ 123456fudan.edu.cn
                Article
                10.3389/fonc.2024.1410110
                11335555
                39169938
                87a7b923-da25-4d20-a8bd-fe0c3e308a07
                Copyright © 2024 Cui, Xuan, Liao and Wang

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 31 March 2024
                : 17 July 2024
                Page count
                Figures: 5, Tables: 3, Equations: 0, References: 75, Pages: 13, Words: 5489
                Funding
                The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The study was supported by the National Natural Science Foundation of China (82072951 to YW), the Science and Technology Commission of Shanghai Municipality (22Y21900100 to YW), and the Shanghai Anticancer Association (SACA-AX202213 to YW).
                Categories
                Oncology
                Original Research
                Custom metadata
                Head and Neck Cancer

                Oncology & Radiotherapy
                papillary thyroid cancer,circulating biomarkers,microrna,cancer diagnosis,ngs

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