MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach
Gastroesophageal adenocarcinoma (GEA) patients with the microsatellite instability (MSI) subtype emerged as optimal candidates for immunotherapy. To date, immunohistochemistry (IHC) is the gold standard for MSI assessment in formalin-fixed paraffin-embedded (FFPE) specimens. However, IHC, although u...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7244 |
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| author | Elisa Boldrin Maria Assunta Piano Rita Alfieri Marcodomenico Mazza Loretta Vassallo Antonio Scapinello Pierluigi Pilati Matteo Curtarello |
| author_facet | Elisa Boldrin Maria Assunta Piano Rita Alfieri Marcodomenico Mazza Loretta Vassallo Antonio Scapinello Pierluigi Pilati Matteo Curtarello |
| author_sort | Elisa Boldrin |
| collection | DOAJ |
| description | Gastroesophageal adenocarcinoma (GEA) patients with the microsatellite instability (MSI) subtype emerged as optimal candidates for immunotherapy. To date, immunohistochemistry (IHC) is the gold standard for MSI assessment in formalin-fixed paraffin-embedded (FFPE) specimens. However, IHC, although useful for diagnostic typing, cannot be used to analyze cell-free DNA (cfDNA) in liquid biopsy, a tool that could overcome tumor heterogeneity and enable longitudinal monitoring. In order to find an alternative diagnostic method to IHC, we analyzed 86 retrospective GEAs FFPE samples with multiplex PCR. Moreover, to verify the feasibility of MSI detection in liquid biopsy, cfDNA samples of five patients that resulted in having MSI in a prospective cohort of 35 patients were evaluated by multiplex PCR, real-time PCR and droplet digital PCR (ddPCR). Analysis of FFPE showed 100% concordance between multiplex PCR and IHC (Cohen’s Kappa agreement = 1). On the contrary, only ddPCR was able to detect MSI in cfDNAs of T3/T4 GEA patients. In conclusion, data highlight the molecular analysis as an optimal alternative to IHC for the diagnostic typing and suggest that the ddPCR assay can be considered as the most reliable and promising molecular approach to detect MSI in the cfDNA of GEA patients. |
| first_indexed | 2024-03-10T09:38:19Z |
| format | Article |
| id | doaj.art-80dfaa977cd34060b6398317efadb0fd |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:38:19Z |
| publishDate | 2021-07-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-80dfaa977cd34060b6398317efadb0fd2023-11-22T03:55:25ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214724410.3390/ijms22147244MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular ApproachElisa Boldrin0Maria Assunta Piano1Rita Alfieri2Marcodomenico Mazza3Loretta Vassallo4Antonio Scapinello5Pierluigi Pilati6Matteo Curtarello7Immunology and Molecular Oncology, Veneto Institute of Oncology IOV-IRCCS, Via Gattamelata 64, 35128 Padova, ItalyImmunology and Molecular Oncology, Veneto Institute of Oncology IOV-IRCCS, Via Gattamelata 64, 35128 Padova, ItalyOncological Surgery Unit, Veneto Institute of Oncology IOV-IRCCS, Via dei Carpani 16, 31033 Castelfranco Veneto, ItalyOncological Surgery Unit, Veneto Institute of Oncology IOV-IRCCS, Via dei Carpani 16, 31033 Castelfranco Veneto, ItalyPathology Unit, Veneto Institute of Oncology IOV-IRCCS, Via dei Carpani 16, 31033 Castelfranco Veneto, ItalyPathology Unit, Veneto Institute of Oncology IOV-IRCCS, Via dei Carpani 16, 31033 Castelfranco Veneto, ItalyOncological Surgery Unit, Veneto Institute of Oncology IOV-IRCCS, Via dei Carpani 16, 31033 Castelfranco Veneto, ItalyImmunology and Molecular Oncology, Veneto Institute of Oncology IOV-IRCCS, Via Gattamelata 64, 35128 Padova, ItalyGastroesophageal adenocarcinoma (GEA) patients with the microsatellite instability (MSI) subtype emerged as optimal candidates for immunotherapy. To date, immunohistochemistry (IHC) is the gold standard for MSI assessment in formalin-fixed paraffin-embedded (FFPE) specimens. However, IHC, although useful for diagnostic typing, cannot be used to analyze cell-free DNA (cfDNA) in liquid biopsy, a tool that could overcome tumor heterogeneity and enable longitudinal monitoring. In order to find an alternative diagnostic method to IHC, we analyzed 86 retrospective GEAs FFPE samples with multiplex PCR. Moreover, to verify the feasibility of MSI detection in liquid biopsy, cfDNA samples of five patients that resulted in having MSI in a prospective cohort of 35 patients were evaluated by multiplex PCR, real-time PCR and droplet digital PCR (ddPCR). Analysis of FFPE showed 100% concordance between multiplex PCR and IHC (Cohen’s Kappa agreement = 1). On the contrary, only ddPCR was able to detect MSI in cfDNAs of T3/T4 GEA patients. In conclusion, data highlight the molecular analysis as an optimal alternative to IHC for the diagnostic typing and suggest that the ddPCR assay can be considered as the most reliable and promising molecular approach to detect MSI in the cfDNA of GEA patients.https://www.mdpi.com/1422-0067/22/14/7244gastroesophageal adenocarcinoma (GEA)microsatellite instability (MSI)cell-free DNA (cfDNA)liquid biopsy |
| spellingShingle | Elisa Boldrin Maria Assunta Piano Rita Alfieri Marcodomenico Mazza Loretta Vassallo Antonio Scapinello Pierluigi Pilati Matteo Curtarello MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach International Journal of Molecular Sciences gastroesophageal adenocarcinoma (GEA) microsatellite instability (MSI) cell-free DNA (cfDNA) liquid biopsy |
| title | MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach |
| title_full | MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach |
| title_fullStr | MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach |
| title_full_unstemmed | MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach |
| title_short | MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach |
| title_sort | msi analysis in solid and liquid biopsies of gastroesophageal adenocarcinoma patients a molecular approach |
| topic | gastroesophageal adenocarcinoma (GEA) microsatellite instability (MSI) cell-free DNA (cfDNA) liquid biopsy |
| url | https://www.mdpi.com/1422-0067/22/14/7244 |
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