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Leveraging colorectal cancer (CRC) as a model, we explored the influence of computational parameters on mutational signature analyses across 230 CRC cell lines and 152 CRC patients. Results were validated in three independent datasets: 483 endometrial cancer patients stratified by mismatch repair (MMR) status, 35 lung cancer patients by smoking status and 12 patient-derived organoids (PDOs) annotated for colibactin exposure. Assessing various bioinformatic tools, reference datasets and input data sizes including whole genome sequencing, whole exome sequencing and a pan-cancer gene panel, we demonstrated significant variability in the results. We report that the use of distinct algorithms and references led to statistically different results, highlighting how arbitrary choices may induce variability in the mutational signature contributions. Furthermore, we found a differential contribution of mutational signatures between coding and intergenic regions and defined the minimum number of somatic variants required for reliable mutational signature assignment. To facilitate the identification of the most suitable workflows, we developed Comparative Mutational Signature analysis on Coding and Extragenic Regions (CoMSCER), a bioinformatic tool which allows researchers to easily perform comparative mutational signature analysis by coupling the results from several tools and public reference datasets and to assess mutational signature contributions in coding and non-coding genomic regions. In conclusion, our study provides a comparative framework to elucidate the impact of distinct computational workflows on mutational signatures.<\/jats:p>","DOI":"10.1093\/bib\/bbae249","type":"journal-article","created":{"date-parts":[[2024,5,24]],"date-time":"2024-05-24T07:37:29Z","timestamp":1716536249000},"source":"Crossref","is-referenced-by-count":10,"title":["Mutational signatures of colorectal cancers according to distinct computational workflows"],"prefix":"10.1093","volume":"25","author":[{"given":"Paolo","family":"Battuello","sequence":"first","affiliation":[{"name":"Department of Oncology , Molecular Biotechnology Center, , Piazza Nizza 44, 10126, Turin , Italy"},{"name":"University of Turin , Molecular Biotechnology Center, , Piazza Nizza 44, 10126, Turin , Italy"},{"name":"Genomics of Cancer and Targeted Therapies Unit , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"},{"name":"The AIRC Institute of Molecular Oncology , IFOM ETS, , Via Adamello 16, 20139, Milan , 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Targeted Therapies Unit , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"},{"name":"The AIRC Institute of Molecular Oncology , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Di Nicolantonio","sequence":"additional","affiliation":[{"name":"Department of Oncology , Molecular Biotechnology Center, , Piazza Nizza 44, 10126, Turin , Italy"},{"name":"University of Turin , Molecular Biotechnology Center, , Piazza Nizza 44, 10126, Turin , Italy"},{"name":"Candiolo Cancer Institute , FPO - IRCCS, Strada Provinciale 142 - km 3.95, 10060, Candiolo, Turin , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Bardelli","sequence":"additional","affiliation":[{"name":"Department of Oncology , Molecular Biotechnology Center, , Piazza Nizza 44, 10126, Turin , Italy"},{"name":"University of Turin , Molecular Biotechnology Center, , Piazza Nizza 44, 10126, Turin , Italy"},{"name":"Genomics of Cancer and Targeted Therapies Unit , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"},{"name":"The AIRC Institute of Molecular Oncology , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5511-1555","authenticated-orcid":false,"given":"Giovanni","family":"Crisafulli","sequence":"additional","affiliation":[{"name":"Genomics of Cancer and Targeted Therapies Unit , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"},{"name":"The AIRC Institute of Molecular Oncology , IFOM ETS, , Via Adamello 16, 20139, Milan , Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,5,23]]},"reference":[{"issue":"7463","key":"2024052406004550700_ref1","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1038\/nature12477","article-title":"Signatures of mutational processes in human 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