FOCUS4 biomarker laboratories: from the benefits to the practical and logistical issues faced during 6 years of centralised testing.

Autor:	Richman SD; Leeds Institute on Medical Research, University of Leeds, Leeds, UK s.d.richman@leeds.ac.uk., Hemmings G; Leeds Institute on Medical Research, University of Leeds, Leeds, UK., Roberts H; All Wales Molecular Genetics Laboratory, All Wales Medical Genetics Service, University Hospital of Wales, Cardiff, UK., Gallop N; Leeds Institute on Medical Research, University of Leeds, Leeds, UK., Dodds R; All Wales Molecular Genetics Laboratory, All Wales Medical Genetics Service, University Hospital of Wales, Cardiff, UK., Wilkinson L; Leeds Institute on Medical Research, University of Leeds, Leeds, UK., Davis J; Leeds Institute on Medical Research, University of Leeds, Leeds, UK., White R; All Wales Molecular Genetics Laboratory, All Wales Medical Genetics Service, University Hospital of Wales, Cardiff, UK., Yates E; MRC Clinical Trials Unit at UCL, London, UK., Jasani B; TARGOS Molecular Pathology GmbH, Kassel, Germany., Brown L; MRC Clinical Trials Unit at UCL, London, UK., Maughan TS; MRC Oxford Institute for Radiation Oncology, University of Oxford, Oxford, UK., Butler R; All Wales Molecular Genetics Laboratory, All Wales Medical Genetics Service, University Hospital of Wales, Cardiff, UK., Quirke P; Leeds Institute on Medical Research, University of Leeds, Leeds, UK., Adams R; Velindre Cancer Centre, Cardiff University, Cardiff, UK.
Jazyk:	angličtina
Zdroj:	Journal of clinical pathology [J Clin Pathol] 2023 Aug; Vol. 76 (8), pp. 548-554. Date of Electronic Publication: 2022 Mar 07.
DOI:	10.1136/jclinpath-2022-208233
Abstrakt:	Aims: FOCUS4 was a phase II/III umbrella trial, recruiting patients with advanced or metastatic colorectal cancer, between 2014 and 2020. Molecular profiling of patients' formalin-fixed, paraffin-embedded tumour blocks was undertaken at two centralised biomarker laboratories (Leeds and Cardiff), and the results fed directly to the Medical Research Council Clinical Trials Unit, and used for subsequent randomisation. Here the laboratories discuss their experiences. Methods: Following successful tumour content assessment, blocks were sectioned for DNA extraction and immunohistochemistry (IHC). Pyrosequencing was initially used to determine tumour mutation status (KRAS, NRAS, BRAF and PIK3CA), then from 2018 onwards, next-generation sequencing was employed to allow the inclusion of TP53. Protein expression of MLH1, MSH2, MSH6, PMS2 and pTEN was determined by IHC. An interlaboratory comparison programme was initiated, allowing sample exchanges, to ensure continued assay robustness. Results: 1291 tumour samples were successfully analysed. Assay failure rates were very low; 1.9%-3.3% for DNA sequencing and 0.9%-1.3% for IHC. Concordance rates of > 98% were seen for the interlaboratory comparisons, where a result was obtained by both laboratories. Conclusions: Practical and logistical problems were identified, including poor sample quality and difficulties with sample anonymisation. The often last-minute receipt of a sample for testing and a lack of integration with National Health Service mutation analysis services were challenging. The laboratories benefitted from both pretrial validations and interlaboratory comparisons, resulting in robust assay development and provided confidence during the implementation of new sequencing technologies. We conclude that our centralised approach to biomarker testing in FOCUS4 was effective and successful. Competing Interests: Competing interests: None declared. (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY. Published by BMJ.)
Databáze:	MEDLINE
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