Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Droplet Digital PCR Development To Quantify the DNA Meth-Ylation Levels of SEPT9 and SHOX2 in Plasma From Patients With Head and Neck Squamous Cell Carcinoma

Ilaria Grossi
ORCID logo ,
Claudia Assoni
,
Luigi Lorini
,
Davide Smussi
,
Cristina Gurizzan
,
Salvatore Grisanti
,
Alberto Paderno
ORCID logo ,
Davide Mattavelli
ORCID logo ,
Cesare Piazza
,
Iulia Andreea Pelisenco
,
Giuseppina De Petro
ORCID logo ,
Alessandro Salvi
* ORCID logo ,
Paolo Bossi
Version 1 : Received: 5 May 2023 / Approved: 6 May 2023 / Online: 6 May 2023 (08:59:09 CEST)

How to cite: Grossi, I.; Assoni, C.; Lorini, L.; Smussi, D.; Gurizzan, C.; Grisanti, S.; Paderno, A.; Mattavelli, D.; Piazza, C.; Pelisenco, I. A.; De Petro, G.; Salvi, A.; Bossi, P. Droplet Digital PCR Development To Quantify the DNA Meth-Ylation Levels of SEPT9 and SHOX2 in Plasma From Patients With Head and Neck Squamous Cell Carcinoma. Preprints 2023, 2023050397. https://doi.org/10.20944/preprints202305.0397.v1 Grossi, I.; Assoni, C.; Lorini, L.; Smussi, D.; Gurizzan, C.; Grisanti, S.; Paderno, A.; Mattavelli, D.; Piazza, C.; Pelisenco, I. A.; De Petro, G.; Salvi, A.; Bossi, P. Droplet Digital PCR Development To Quantify the DNA Meth-Ylation Levels of SEPT9 and SHOX2 in Plasma From Patients With Head and Neck Squamous Cell Carcinoma. Preprints 2023, 2023050397. https://doi.org/10.20944/preprints202305.0397.v1

Abstract

Head and neck squamous cell carcinomas (HNSCCs) develop from the mucosa of the oral cavity, pharynx, and larynx. The methylation levels of Septin 9 (SEPT9) and short stature homeobox 2 (SHOX2) genes in ccfDNA are considered pan-cancer biomarkers and have shown prognostic value in preliminary reports in HNSCC. Liquid biopsy is a non-invasive procedure that collects tumor-derived molecules including circulating cell-free DNA (ccfDNA). Here, we developed a ddPCR-based assay to detect the DNA methylation levels of plasma circulating SEPT9 and SHOX2 in patients with HNSCC. We first set up the assay on commercial methylated and un-methylated DNA. Then, the dynamic changes of the methylation levels of SEPT9 and SHOX2 were quantified in 20 patients during the follow-up. The results highlighted: i) the capability of the ddPCR-based assay in detecting very low copies of methylated molecules; ii) the significant decrease in methylation levels of SEPT9 and SHOX2 in plasma of HNSCC patients at the first time points of follow-up respect to T0; iii) a different trend of longitudinally DNA methylation variations associable to the clinicopathological features of the patients. The absolute quantification of the methylation levels of SEPT9 and SHOX2 in HNSCC may be used for risk stratification and disease monitoring.

Keywords

liquid biopsy; ddPCR; DNA methylation; cell free DNA; HNSCC

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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