Laboratory-Developed NGS Panel for Characterization of Advanced Colorectal Cancer, Real-World Data from a Tertiary Referral Centre.
4 affiliations
- 1Biobank of Research, IRCCS Azienda Ospedaliero-Universitaria di Bologna, 40138 Bologna, Italy.
- 2Solid Tumor Molecular Pathology Laboratory, IRCCS Azienda Ospedaliero-Universitaria di Bologna, 40138 Bologna, Italy.
- 3Department of Medical and Surgical Sciences (DIMEC), University of Bologna, 40138 Bologna, Italy.
- 4Department of Pharmacy and Biotechnology (FaBiT), University of Bologna, 40138 Bologna, Italy.
Abstract
Background/Objectives: Colorectal cancer (CRC) is one of the most frequently diagnosed malignancies and a leading cause of cancer-related mortality worldwide. Its development and progression are driven by a complex interplay of genetic and epigenetic alterations that contribute to marked biological heterogeneity and variable clinical outcomes. Comprehensive molecular profiling is therefore essential for accurately characterizing CRC and guiding personalized therapeutic strategies. In this context, next-generation sequencing (NGS) has emerged as a powerful tool, enabling high-throughput, simultaneous detection of multiple clinically relevant genomic alterations in a single assay. Methods: We report our experience with a laboratory-designed NGS panel for the routine assessment of advanced-stage CRC. A total of 611 colorectal cancer specimens were analyzed using a laboratory-developed multi-gene NGS panel targeting 229 amplicons across 25 oncogenes and tumor suppressor genes. Results: The panel demonstrated analytical feasibility, with unsuccessful results observed in only 1.3% of the analyzed samples, due to low-quality/quantity DNA. RAS genes (KRAS and NRAS, altered in 51.2% and 2.7% of cases, respectively), together with BRAF (10.6%), PIK3CA (16.1%), SMAD4 (11.8%), and TP53 (64.3%), represented the most frequently altered molecular markers in the studied cohort. The frequency of major actionable genetic alterations in CRC was comparable between primary (n = 482) and metastatic (n = 121) lesions, suggesting similar mutation frequencies across the groups. Conclusions: This study presents a laboratory-developed multi-gene NGS panel specifically designed for molecular profiling of CRC in routine clinical practice.
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BibTeX
@article{paola2026laboratorydeveloped,
title = {{Laboratory-Developed NGS Panel for Characterization of Advanced Colorectal Cancer, Real-World Data from a Tertiary Referral Centre.}},
author = {Floriana Jessica Di Paola and Giulia Calafato and Thais Maloberti and Laura Poppi and Giulia Ciccimarra and Sara Coluccelli and Viviana Sanza and Elisa Gruppioni and Annalisa Altimari and Antonio De Leo and Dario de Biase and Giovanni Tallini},
journal = {Diagnostics (Basel, Switzerland)},
year = {2026},
month = sep,
doi = {10.3390/diagnostics16193091},
url = {https://europepmc.org/article/MED/42882247},
note = {CC BY},
}
RIS
TY - JOUR
TI - Laboratory-Developed NGS Panel for Characterization of Advanced Colorectal Cancer, Real-World Data from a Tertiary Referral Centre.
AU - Floriana Jessica Di Paola
AU - Giulia Calafato
AU - Thais Maloberti
AU - Laura Poppi
AU - Giulia Ciccimarra
AU - Sara Coluccelli
AU - Viviana Sanza
AU - Elisa Gruppioni
AU - Annalisa Altimari
AU - Antonio De Leo
AU - Dario de Biase
AU - Giovanni Tallini
PY - 2026
DA - 2026/09/24
JO - Diagnostics (Basel, Switzerland)
DO - 10.3390/diagnostics16193091
UR - https://europepmc.org/article/MED/42882247
AB - Background/Objectives: Colorectal cancer (CRC) is one of the most frequently diagnosed malignancies and a leading cause of cancer-related mortality worldwide. Its development and progression are driven by a complex interplay of genetic and epigenetic alterations that contribute to marked biological heterogeneity and variable clinical outcomes. Comprehensive molecular profiling is therefore essential for accurately characterizing CRC and guiding personalized therapeutic strategies. In this context, next-generation sequencing (NGS) has emerged as a powerful tool, enabling high-throughput, simultaneous detection of multiple clinically relevant genomic alterations in a single assay. Methods: We report our experience with a laboratory-designed NGS panel for the routine assessment of advanced-stage CRC. A total of 611 colorectal cancer specimens were analyzed using a laboratory-developed multi-gene NGS panel targeting 229 amplicons across 25 oncogenes and tumor suppressor genes. Results: The panel demonstrated analytical feasibility, with unsuccessful results observed in only 1.3% of the analyzed samples, due to low-quality/quantity DNA. RAS genes (KRAS and NRAS, altered in 51.2% and 2.7% of cases, respectively), together with BRAF (10.6%), PIK3CA (16.1%), SMAD4 (11.8%), and TP53 (64.3%), represented the most frequently altered molecular markers in the studied cohort. The frequency of major actionable genetic alterations in CRC was comparable between primary (n = 482) and metastatic (n = 121) lesions, suggesting similar mutation frequencies across the groups. Conclusions: This study presents a laboratory-developed multi-gene NGS panel specifically designed for molecular profiling of CRC in routine clinical practice.
SN - 2075-4418
ER -