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前列腺癌病例内部概述及来自FFPE组织的NGS数据质量

Internal Overview of Prostatic Cancer Cases and Quality of and NGS Data from the FFPE Tissue.

作者信息

Antolini Enrica, Filosa Alessandra, Santoni Matteo, Antaldi Elena, Bartoli Elisa, Sierchio Lidia, Giantomassi Federica, Mandolesi Alessandra, Goteri Gaia

机构信息

Department of Biomedical Science and Public Health, Università Politecnica delle Marche, 60126 Ancona, Italy.

Anatomic Pathology Unit, Azienda Ospedaliero Universitaria delle Marche, 60126 Ancona, Italy.

出版信息

Diagnostics (Basel). 2024 Sep 18;14(18):2067. doi: 10.3390/diagnostics14182067.

Abstract

Comprehensive genomic profiling (CGP) has gained an important role in patients with advanced prostate cancer following the introduction of PARP inhibitors in daily clinical practice. Here, we report an overview of CGP results, specifically of BRCA1 and BRCA2 HRD-repair system genes, from patients with prostate cancer analyzed in our institution, and we compare our results with those available from more recent scientific literature. The study cohort consisted of 70 patients. Somatic DNA was extracted from Formalin-Fixed Paraffin-Embedded (FFPE) tissue using a MagCore Genomic DNA FFPE One-Step Kit for MagCore System. The DNA was quantified by EasyPGX Real-Time qPCR and EasyPGX Analysis Software (version 4.0.13). Tissue somatic DNA libraries were prepared with Myriapod NGS BRCA1-2 panel-NG035 and sequenced in a Mi-Seq System. The sequence alignment in hg19 and the variant calling were performed using Myriapod NGS Data Analysis Software version 5.0.8 NG900-SW 5.0.8 with a software detection limit (LoD) of 95%. Variants with a coverage of 500 and VAF% ≥ 5 were evaluated. Tumor tissue NGS was unsuccessful in 46/70 patients (66%). Mutations of the gene were detected in 4 of the samples: (1) ex10 c.1244A>G p.His415Arg VAF = 51.03%; (2) ex11 c.5946delT p.Ser1982fs VAF = 72.1%; (3) ex11 c.3302A>G p.His1101Arg VAF = 52.9%; and (4) ex11 c.3195_3198delTAAT p.Asn1066fs VAF = 51.1%. The results from our internal overview seem to support the data and to confirm the performance of the technical issues reported in the literature. Considering the advanced age of our patients, with 84% of men over the age of 65, the application of alternative and less invasive procedures such as liquid biopsy, could be a more suitable solution for some cases.

摘要

在日常临床实践中引入聚二磷酸腺苷核糖聚合酶(PARP)抑制剂后,综合基因组分析(CGP)在晚期前列腺癌患者中发挥了重要作用。在此,我们报告了在我们机构分析的前列腺癌患者的CGP结果概述,特别是BRCA1和BRCA2同源重组缺陷(HRD)修复系统基因的结果,并将我们的结果与最新科学文献中的结果进行比较。研究队列由70名患者组成。使用用于MagCore系统的MagCore基因组DNA福尔马林固定石蜡包埋(FFPE)一步法试剂盒从FFPE组织中提取体细胞DNA。通过EasyPGX实时定量聚合酶链反应(qPCR)和EasyPGX分析软件(版本4.0.13)对DNA进行定量。用Myriapod NGS BRCA1-2检测板-NG035制备组织体细胞DNA文库,并在Mi-Seq系统中进行测序。使用Myriapod NGS数据分析软件版本5.0.8 NG900-SW 5.0.8在hg19中进行序列比对和变异检测,软件检测限(LoD)为95%。评估覆盖度为500且变异等位基因频率(VAF)%≥5的变异。70例患者中有46例(66%)肿瘤组织的下一代测序(NGS)未成功。在4个样本中检测到该基因的突变:(1)外显子10 c.1244A>G p.His415Arg,VAF = 51.03%;(2)外显子11 c.5946delT p.Ser1982fs,VAF = 72.1%;(3)外显子11 c.3302A>G p.His1101Arg,VAF = 52.9%;以及(4)外显子11 c.3195_3198delTAAT p.Asn1066fs,VAF = 51.1%。我们内部概述的结果似乎支持这些数据,并证实了文献中报道的技术问题的表现。考虑到我们患者的高龄,65岁以上男性占84%,对于某些病例,应用替代的、侵入性较小的方法,如液体活检,可能是更合适的解决方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ca21/11431729/1028afb4d138/diagnostics-14-02067-g001.jpg

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