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Multi-gene custom panels for the characterisation of metastatic colorectal carcinoma in clinical practice: express the role of PIK3CA mutations
  1. Dario de Biase1,2,
  2. Umberto Malapelle3,
  3. Antonio De Leo2,4,
  4. Thais Maloberti4,
  5. Michela Visani4,
  6. Pasquale Pisapia3,
  7. Giorgia Acquaviva2,4,
  8. Francesco Pepe5,
  9. Gianluca Russo3,
  10. Antonino Iaccarino3,
  11. Annalisa Pession1,2,
  12. Giovanni Tallini2,4,
  13. Giancarlo Troncone3
  1. 1 Department of Pharmacy and Biotechnology, Molecular Diagnostic Unit, University of Bologna, Bologna, Italy
  2. 2 Division of Molecular Pathology Laboratory, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy
  3. 3 Department of Public Health, University Federico II of Naples, Napoli, Italy
  4. 4 Department of Medicine (Dipartimento di Medicina Specialistica, Diagnostica e Sperimentale), Molecular Diagnostic Unit, University of Bologna, Azienda USL di Bologna, Bologna, Italy
  5. 5 Sanità Pubblica, University of Naples Federico II, Napoli, Italy
  1. Correspondence to Dr Dario de Biase, Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy; dario.debiase{at}; Dr Umberto Malapelle; umberto.malapelle{at}


Aims In metastatic colorectal carcinomas (mCRC), RAS/RAF genes mutations are first tested to determine the eligibility for anti-EGFR (Epidermal Growth Factor Receptor) therapy in combination with conventional cytotoxic agents. Recent advancements in next-generation sequencing (NGS) have highlighted the potential of multi-gene panels. This multi-gene analysis may provide useful information for the molecular characterisation of mCRC, other than the status of RAS/RAF genes. Aim of this study was to evaluate the feasibility of two NGS custom multi-gene panels in the characterisation of CRC cases and evaluating the relevance of PIK3CA mutation in a routine cohort of consecutive CRC cases.

Methods A total of 961 formalin-fixed and paraffin-embedded specimens from two medical centres (Bologna and Naples) were analysed using two lab-developed NGS multi-gene panels.

Results KRAS mutations (56.2%) were the more frequent alterations observed in our cohort. Intriguingly, PIK3CA mutations were more frequent (16.8%) than variants observed in the other two genes nowadays analysed in CRC clinical practice (NRAS and BRAF, 4.2% and 9.6%, respectively). Moreover, in more than 10% of samples, coexistent mutations were detected in our cohort of CRC.

Conclusions Our study demonstrates the feasibility and efficacy of lab-developed targeted multi-gene NGS panels in the clinical practice of CRC. Moreover, the data lead to hypothesise that PIK3CA mutations, together with those of RAS/BRAF, worth to be further investigated in clinical CRC specimens.

  • colorectal cancer
  • pathology
  • molecular
  • oncogenes
  • DNA
  • diagnostic techniques and procedures

Data availability statement

All data relevant to the study are included in the article or uploaded as supplementary information.

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Data availability statement

All data relevant to the study are included in the article or uploaded as supplementary information.

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  • GT and GT are joint senior authors.

  • Handling editor Runjan Chetty.

  • Twitter @UmbertoMalapel1, @PasqualePisapia

  • DdB and UM contributed equally.

  • Contributors Conceptualisation, DdB and UM; methodology, DdB, UM, MV, PP, GA, FP, ADL, TM, GR, AI; formal analysis, DdB, UM, PP, FP; data curation, PP, GA, FP, ADL, GR, TM, AP; writing—original draft preparation, DdB, UM; supervision, AP, GTallini, GTroncone; project administration, GTallini, GTroncone; funding acquisition, GTallini, GTroncone. All authors have read and agreed to the published version of the manuscript.

  • Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.

  • Competing interests UM reports personal fees (as a speaker or advisor) from Boehringer Ingelheim, ThermoFisher Scientific, AstraZeneca, Roche, MSD (Merck Sharp & Dohme), Amgen and Merck, Diaceutics and Eli Lilly, which are unrelated to the current work.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.