L. P. Priesterbach-Ackley, Joyce van Kuik, B. Tops, A. Lasorella, A. Iavarone, Wim van Hecke, Pierre A. Robe, Pieter Wesseling, Wendy W J de Leng
{"title":"RT-PCR assay to detect FGFR3::TACC3 fusions in formalin-fixed, paraffin-embedded glioblastoma samples","authors":"L. P. Priesterbach-Ackley, Joyce van Kuik, B. Tops, A. Lasorella, A. Iavarone, Wim van Hecke, Pierre A. Robe, Pieter Wesseling, Wendy W J de Leng","doi":"10.1093/nop/npad081","DOIUrl":null,"url":null,"abstract":"\n \n \n One targeted treatment option for IDH-wildtype glioblastoma focuses on tumors with FGFR3::TACC3 fusions. FGFR3::TACC3 fusion detection can be challenging, as targeted RNA next generation sequencing is not routinely performed and immunohistochemistry is an imperfect surrogate marker. Fusion status can be determined using RT-PCR on fresh frozen (FF) material, but sometimes only formalin-fixed, paraffin embedded (FFPE) tissue is available.\n \n \n \n To develop an RT-PCR assay to determine FGFR3::TACC3 status in FFPE glioblastoma samples.\n \n \n \n Twelve tissue micro-arrays with 353 historical glioblastoma samples were immunohistochemically stained for FGFR3. Samples with overexpression of FGFR3 (n=13) were subjected to FGFR3::TACC3 RT-PCR on FFPE, using 5 primer sets for detection of 5 common fusion variants. Fusion negative samples were additionally analyzed with NGS (n=6), FGFR3 FISH (n=6) and RNA sequencing (n=5).\n \n \n \n Using RT-PCR on FFPE material of the 13 samples with FGFR3 overexpression, we detected an FGFR3::TACC3 fusion in 7 samples, covering 3 different fusion variants. For 5 of these FF was available, and the presence of the fusion was confirmed through RT-PCR on FF. With RNA-sequencing one additional sample was found to harbor a FGFR3::TACC3 fusion (variant not covered by current RT-PCR for FFPE). The frequency of FGFR3::TACC3 fusion in this cohort was 9/353 (2,5%).\n \n \n \n RT-PCR for FGFR3::TACC3 fusions can successfully be performed on FFPE material, with a specificity of 100% and (due to limited primer sets) a sensitivity of 83,3%. This assay allows for the identification of potential targeted treatment options when only formalin-fixed tissue is available.\n","PeriodicalId":506567,"journal":{"name":"Neuro-Oncology Practice","volume":"49 24","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Neuro-Oncology Practice","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/nop/npad081","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
One targeted treatment option for IDH-wildtype glioblastoma focuses on tumors with FGFR3::TACC3 fusions. FGFR3::TACC3 fusion detection can be challenging, as targeted RNA next generation sequencing is not routinely performed and immunohistochemistry is an imperfect surrogate marker. Fusion status can be determined using RT-PCR on fresh frozen (FF) material, but sometimes only formalin-fixed, paraffin embedded (FFPE) tissue is available.
To develop an RT-PCR assay to determine FGFR3::TACC3 status in FFPE glioblastoma samples.
Twelve tissue micro-arrays with 353 historical glioblastoma samples were immunohistochemically stained for FGFR3. Samples with overexpression of FGFR3 (n=13) were subjected to FGFR3::TACC3 RT-PCR on FFPE, using 5 primer sets for detection of 5 common fusion variants. Fusion negative samples were additionally analyzed with NGS (n=6), FGFR3 FISH (n=6) and RNA sequencing (n=5).
Using RT-PCR on FFPE material of the 13 samples with FGFR3 overexpression, we detected an FGFR3::TACC3 fusion in 7 samples, covering 3 different fusion variants. For 5 of these FF was available, and the presence of the fusion was confirmed through RT-PCR on FF. With RNA-sequencing one additional sample was found to harbor a FGFR3::TACC3 fusion (variant not covered by current RT-PCR for FFPE). The frequency of FGFR3::TACC3 fusion in this cohort was 9/353 (2,5%).
RT-PCR for FGFR3::TACC3 fusions can successfully be performed on FFPE material, with a specificity of 100% and (due to limited primer sets) a sensitivity of 83,3%. This assay allows for the identification of potential targeted treatment options when only formalin-fixed tissue is available.