Moreno Daniel Antunes, da Silva Luciane Sussuchi, Zanon Maicon Fernando, Bonatelli Murilo, de Paula Flávia Escremim, de Medeiros Matsushita Marcus, Teixeira Gustavo Ramos, Santana Iara Viana Vidigal, Saggioro Fabiano, Neder Luciano, Stavale João N, Malheiros Suzana Maria Fleury, Lima Matheus, Hajj Glaucia Noeli Maroso, Garcia-Rivello Hernan, Christiansen Silvia, Nunes Susana, da Costa Maria João Gil, Soares Maria José, Pinheiro Jorge, Junior Carlos Almeida, Mançano Bruna Minniti, Reis Rui Manuel
Molecular Oncology Research Center Dr. Paulo Prata, Barretos Cancer Hospital, Barretos, Brazil.
Molecular Diagnosis Laboratory Dr. Paulo Prata, Barretos Cancer Hospital, Barretos, Brazil.
J Neurooncol. 2022 Mar;157(1):27-35. doi: 10.1007/s11060-022-03965-1. Epub 2022 Feb 15.
Medulloblastoma is the most frequent pediatric malignant brain tumor, and is divided into four main subgroups: WNT, SHH, group 3, and group 4. MYCN amplification is an important medulloblastoma prognostic biomarker. We aimed to molecular classify and predict MYCN amplification in a single assay.
It was included 209 medulloblastomas from 205 patients (Brazil, Argentina, and Portugal), divided into training (n = 50) and validation (n = 159) sets. A nCounter assay was carried out using a custom panel for molecular classification, with additional genes, including MYCN. nSolver 4.0 software and the R environment were used for profiling and MYCN mRNA analysis. MYCN amplification by FISH was performed in 64 cases.
The 205 medulloblastomas were classified in SHH (44.9%), WNT (15.6%), group 3 (18.1%) and group 4 (21.4%). In the training set, MYCN amplification was detected in three SHH medulloblastomas by FISH, which showed significantly higher MYCN mRNA counts than non-FISH amplified cases, and a cutoff for MYCN amplification was established ([Formula: see text] + 4σ = 11,124.3). Applying this threshold value in the validation set, we identified MYCN mRNA counts above the cutoff in three cases, which were FISH validated.
We successfully stratified medulloblastoma molecular subgroups and predicted MYCN amplification using a single nCounter assay without the requirement of additional biological tissue, costs, or bench time.
髓母细胞瘤是最常见的儿童恶性脑肿瘤，分为四个主要亚组：WNT、SHH、3组和4组。MYCN扩增是髓母细胞瘤重要的预后生物标志物。我们旨在通过单一检测对髓母细胞瘤进行分子分类并预测MYCN扩增。
纳入来自205例患者（巴西、阿根廷和葡萄牙）的209例髓母细胞瘤，分为训练集（n = 50）和验证集（n = 159）。使用定制的分子分类检测板进行nCounter检测，该检测板包含包括MYCN在内的其他基因。使用nSolver 4.0软件和R环境进行分析及MYCN mRNA分析。对64例病例进行了FISH检测MYCN扩增。
205例髓母细胞瘤分为SHH（44.9%）、WNT（15.6%）、3组（18.1%）和4组（21.4%）。在训练集中，通过FISH在3例SHH髓母细胞瘤中检测到MYCN扩增，这些病例的MYCN mRNA计数显著高于未通过FISH检测到扩增的病例，并确定了MYCN扩增的临界值（[公式：见正文]+4σ = 11,124.3）。在验证集中应用该阈值，我们在3例病例中鉴定出MYCN mRNA计数高于临界值，这些病例经FISH验证。
我们成功地对髓母细胞瘤分子亚组进行了分层，并使用单一的nCounter检测预测了MYCN扩增，无需额外的生物组织、成本或实验时间。
