Unbiased Detection of Driver Mutations in Extramammary Paget Disease
Unbiased Detection of Driver Mutations in Extramammary Paget Disease
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DOI：
10.1158/1078-0432.ccr-20-3205
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发表时间：
2021-03-15
影响因子：
11.5
通讯作者：
Ogawa, Seishi
中科院分区：
文献类型：
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作者：
Ishida, Yoshihiro;Kakiuchi, Nobuyuki;Ogawa, Seishi
目的：乳房外佩吉特病（EMPD）是一种罕见的皮肤恶性肿瘤，其遗传学改变特征性不强。先前的报道鉴定了染色质重塑基因和PIK 3CA的突变。为了明确地确定EMPD中的驱动突变，我们使用外显子组测序结合靶向测序分析了87个EMPD样品。实验设计：首先，我们使用全外显子组测序分析了37个手术切除的EMPD样品。基于几个计算机分析，我们建立了一个自定义的捕获面板推定的驱动基因，并分析了50个额外的福尔马林固定，石蜡包埋的样本使用靶测序。通过HER 2免疫组织化学评价ERBB 2表达。通过荧光原位杂交进一步分析选择的样品。结果：在外显子组分析中确定了92个突变/样品的中位数。驱动检测算法的联合鉴定ERBB 2、ERBB 3、KMT 2C、TP 53、PIK 3CA、NUP 93、AFDN和CUX 1为可能的驱动突变。拷贝数改变分析显示跨越CDKN 2A的区域反复缺失，ERBB 2反复扩增。ERBB 2、ERBB 3和FGFR 1的扩增/突变表现出相互排斥的趋势。拷贝数改变负荷与复发的可能性相关。突变特征主要由老化和APOBEC激活，缺乏紫外线辐射的证据。HER 2 IHC/荧光原位分析验证了ERBB 2扩增，但检测突变的能力不足。在某些cases.Conclusions：我们的全面，公正的分析表明EMPD的特点是改变涉及PI 3 K-AKT通路的ERBB 2扩增状态的肿瘤异质性。EMPD在分子途径改变和诱变背后的病因学方面与其他皮肤癌不同。
Purpose: Extramammary Paget disease (EMPD) is an uncommon skin malignancy whose genetic alterations are poorly characterized. Previous reports identified mutations in chromatin remodeling genes and PIK3CA. In order to unambiguously determine driver mutations in EMPD, we analyzed 87 EMPD samples using exome sequencing in combination with targeted sequencing.Experimental Design: First, we analyzed 37 EMPD samples that were surgically resected using whole-exome sequencing. Based on several in silico analysis, we built a custom capture panel of putative driver genes and analyzed 50 additional formalin-fixed, paraffin-embedded samples using target sequencing. ERBB2 expression was evaluated by HER2 immunohisotochemistry. Select samples were further analyzed by fluorescence in situ hybridization.Results: A median of 92 mutations/sample was identified in exome analysis. A union of driver detection algorithms identified ERBB2, ERBB3, KMT2C, TP53, PIK3CA, NUP93, AFDN, and CUX1 as likely driver mutations. Copy-number alteration analysis showed regions spanning CDKN2A as recurrently deleted, and ERBB2 as recurrently amplified. ERBB2, ERBB3, and FGFR1 amplification/mutation showed tendency toward mutual exclusivity. Copy-number alteration load was associated with likelihood to recur. Mutational signatures were dominated by aging and APOBEC activation and lacked evidence of ultraviolet radiation. HER2 IHC/fluorescence in situ analysis validated ERBB2 amplification but was underpowered to detect mutations. Tumor heterogeneity in terms of ERBB2 amplification status was observed in some cases.Conclusions: Our comprehensive, unbiased analysis shows EMPD is characterized by alterations involving the PI3K-AKT pathway. EMPD is distinct from other skin cancers in both molecular pathways altered and etiology behind mutagenesis.