Willing Eva-Maria, Vollbrecht Claudia, Vössing Christine, Weist Peggy, Schallenberg Simon, Herbst Johanna M, Schatz Stefanie, Jóri Balázs, Bataillon Guillaume, Harter Philipp, Salutari Vanda, Martin Antonio Gonzáles, Vergote Ignace, Colombo Nicoletta, Roeper Julia, Berg Tobias, Berger Regina, Kah Bettina, Noettrup Trine Jakobi, Falk Markus, Arndt Kathrin, Polten Andreas, Ray-Coquard Isabelle, Selzam Franziska, Pirngruber Judith, Schmidt Stefanie, Hummel Michael, Tiemann Markus, Horst David, Sehouli Jalid, Pujade-Lauraine Eric, Tiemann Katharina, Braicu Elena Ioana, Heukamp Lukas C
Institut für Hämatopathologie Hamburg, 22547 Hamburg, Germany.
Nord-Ostdeutsche Gesellschaft für Gynäkologische Onkologie NOGGO e. V., 13359 Berlin, Germany.
Cancers (Basel). 2023 Jun 30;15(13):3445. doi: 10.3390/cancers15133445.
The worldwide approval of the combination maintenance therapy of olaparib and bevacizumab in advanced high-grade serous ovarian cancer requires complex molecular diagnostic assays that are sufficiently robust for the routine detection of driver mutations in homologous recombination repair (HRR) genes and genomic instability (GI), employing formalin-fixed (FFPE) paraffin-embedded tumor samples without matched normal tissue. We therefore established a DNA-based hybrid capture NGS assay and an associated bioinformatic pipeline that fulfils our institution's specific needs. The assay´s target regions cover the full exonic territory of relevant cancer-related genes and HRR genes and more than 20,000 evenly distributed single nucleotide polymorphism (SNP) loci to allow for the detection of genome-wide allele specific copy number alterations (CNA). To determine GI status, we implemented an %CNA score that is robust across a broad range of tumor cell content (25-85%) often found in routine FFPE samples. The assay was established using high-grade serous ovarian cancer samples for which and mutation status as well as Myriad MyChoice homologous repair deficiency (HRD) status was known. The NOGGO (Northeastern German Society for Gynecologic Oncology) GIS (GI-Score) v1 assay was clinically validated on more than 400 samples of the ENGOT PAOLA-1 clinical trial as part of the European Network for Gynaecological Oncological Trial groups (ENGOT) HRD European Initiative. The "NOGGO GIS v1 assay" performed using highly robust hazard ratios for progression-free survival (PFS) and overall survival (OS), as well a significantly lower dropout rate than the Myriad MyChoice clinical trial assay supporting the clinical utility of the assay. We also provide proof of a modular and scalable routine diagnostic method, that can be flexibly adapted and adjusted to meet future clinical needs, emerging biomarkers, and further tumor entities.
奥拉帕利和贝伐单抗联合维持疗法在晚期高级别浆液性卵巢癌中的全球获批，需要复杂的分子诊断检测方法，这些方法要足够强大，能够在不匹配正常组织的情况下，使用福尔马林固定（FFPE）石蜡包埋肿瘤样本，对同源重组修复（HRR）基因中的驱动突变和基因组不稳定性（GI）进行常规检测。因此，我们建立了一种基于DNA的杂交捕获NGS检测方法以及相关的生物信息学流程，以满足我们机构的特定需求。该检测方法的目标区域覆盖了相关癌症相关基因和HRR基因的全部外显子区域，以及超过20,000个均匀分布的单核苷酸多态性（SNP）位点，以检测全基因组等位基因特异性拷贝数改变（CNA）。为了确定GI状态，我们实施了一个%CNA评分，该评分在常规FFPE样本中常见的广泛肿瘤细胞含量范围（25-85%）内都很稳健。该检测方法是使用高级别浆液性卵巢癌样本建立的，其 和 突变状态以及Myriad MyChoice同源修复缺陷（HRD）状态是已知的。作为欧洲妇科肿瘤试验组网络（ENGOT）HRD欧洲倡议的一部分，NOGGO（德国东北部妇科肿瘤学会）GIS（GI评分）v1检测方法在ENGOT PAOLA-1临床试验的400多个样本上进行了临床验证。使用“NOGGO GIS v1检测方法”得出的无进展生存期（PFS）和总生存期（OS）的风险比非常稳健，并且 dropout率明显低于Myriad MyChoice临床试验检测方法，这支持了该检测方法的临床实用性。我们还提供了一种模块化和可扩展的常规诊断方法的证据，该方法可以灵活调整和适应，以满足未来的临床需求、新出现的生物标志物以及更多的肿瘤实体。
