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dc.contributor.authorOzgencil, Meryem
dc.contributor.authorBarwell, Julian
dc.contributor.authorTischkowitz, Marc
dc.contributor.authorIzatt, Louise
dc.contributor.authorKesterton, Ian
dc.contributor.authorSimpson, Michael
dc.contributor.authorSharpe, Paul
dc.contributor.authorde Sepulveda, Paulo
dc.contributor.authorVoisset, Edwige
dc.contributor.authorSolomon, Ellen
dc.date.accessioned2021-12-22T15:05:29Z
dc.date.available2021-12-22T15:05:29Z
dc.date.issued2021
dc.date.submitted2021-09-01
dc.identifier.issn1932-6203
dc.identifier.otherpone-d-21-28014
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/331687
dc.descriptionFunder: The European Union Scholarship Programme
dc.descriptionFunder: King’s College London
dc.description.abstractEstablishing a universally applicable protocol to assess the impact of BRCA1 variants of uncertain significance (VUS) expression is a problem which has yet to be resolved despite major progresses have been made. The numerous difficulties which must be overcome include the choices of cellular models and functional assays. We hypothesised that the use of induced pluripotent stem (iPS) cells might facilitate the standardisation of protocols for classification, and could better model the disease process. We generated eight iPS cell lines from patient samples expressing either BRCA1 pathogenic variants, non-pathogenic variants, or BRCA1 VUSs. The impact of these variants on DNA damage repair was examined using a ɣH2AX foci formation assay, a Homologous Repair (HR) reporter assay, and a chromosome abnormality assay. Finally, all lines were tested for their ability to differentiate into mammary lineages in vitro. While the results obtained from the two BRCA1 pathogenic variants were consistent with published data, some other variants exhibited differences. The most striking of these was the BRCA1 variant Y856H (classified as benign), which was unexpectedly found to present a faulty HR repair pathway, a finding linked to the presence of an additional variant in the ATM gene. Finally, all lines were able to differentiate first into mammospheres, and then into more advanced mammary lineages expressing luminal- or basal-specific markers. This study stresses that BRCA1 genetic analysis alone is insufficient to establish a reliable and functional classification for assessment of clinical risk, and that it cannot be performed without considering the other genetic aberrations which may be present in patients. The study also provides promising opportunities for elucidating the physiopathology and clinical evolution of breast cancer, by using iPS cells.
dc.languageen
dc.publisherPublic Library of Science (PLoS)
dc.subjectResearch Article
dc.subjectBiology and life sciences
dc.subjectMedicine and health sciences
dc.subjectResearch and analysis methods
dc.titleAssessing BRCA1 activity in DNA damage repair using human induced pluripotent stem cells as an approach to assist classification of BRCA1 variants of uncertain significance.
dc.typeArticle
dc.date.updated2021-12-22T15:05:28Z
prism.issueIdentifier12
prism.publicationNamePLoS One
prism.volume16
dc.identifier.doi10.17863/CAM.79137
dcterms.dateAccepted2021-11-18
rioxxterms.versionofrecord10.1371/journal.pone.0260852
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
datacite.contributor.supervisoreditor: Galli, Alvaro
dc.contributor.orcidTischkowitz, Marc [0000-0002-7880-0628]
dc.contributor.orcidde Sepulveda, Paulo [0000-0001-8295-5414]
dc.contributor.orcidVoisset, Edwige [0000-0002-0943-4847]
dc.identifier.eissn1932-6203
pubs.funder-project-idBreast Cancer Now (2015NovPR609)
pubs.funder-project-idBlood Cancer UK (13043)
cam.issuedOnline2021-12-02


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