Integrated approach to functional analysis of an ERBB2 variant of unknown significance detected by a cancer gene panel test.
Integrated approach to functional analysis of an ERBB2 variant of unknown significance detected by a cancer gene panel test.
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DOI：
10.1007/s13402-021-00656-3
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Sueoka-Aragane N
中科院分区：
文献类型：
--
作者：
Harada Y;Sato A;Araki M;Matsumoto S;Isaka Y;Sagae Y;Abe T;Aoyagi Y;Sueoka E;Okuno Y;Kimura S;Sueoka-Aragane N
未知意义的变异（VUS）的处理是基于NGS的癌症基因组检测临床应用中的一个重要问题。我们检测到一个新的ERBB 2胞外结构域VUS，c.1157A > G p。（E401 G），在癌症基因组测试中。由于ERBB 2胞外域（ECD）变体的激活机制尚未完全清楚，我们的目的是阐明ERBB 2 E401 G的这些机制和生物学功能。 选择ERBB 2 E401 G作为VUS进行分析，因为多种软件工具预测其致病性。我们制备了具有E401 G变体的ERBB 2表达载体以及具有S310 F和E321 G的载体，这些载体已知是激活突变。基于野生型ERBB 2或突变型ERBB 2在没有ERBB 2扩增或变体的细胞系中的表达，我们评估了人表皮生长因子受体2和相关蛋白的磷酸化，并利用分子动力学（MD）模拟研究了变体赋予的机制。研究了ERBB 2 E401 G的体内外生物学效应。我们发现ERBB 2 E401 G以类似于S310 F的方式增强C-末端磷酸化。MD模拟分析显示，这些变体以配体非依赖性方式维持EGFR-HER 2异源二聚体的稳定性。此外，ERBB 2 E401 G转导的细胞显示出体外侵袭能力增加和体内肿瘤生长能力增加。我们的研究结果提供了关于ERBB 2胞外结构域（ECD）变体激活机制的重要信息，并说明了一个模型工作流程，该流程整合了湿和干工作台过程，用于分析癌症基因组检测检测到的VUS。在线版本包含补充材料，可通过10.1007/s13402-021-00656-3获得。
Dealing with variants of unknown significance (VUS) is an important issue in the clinical application of NGS-based cancer gene panel tests. We detected a novel ERBB2 extracellular domain VUS, c.1157A > G p.(E401G), in a cancer gene panel test. Since the mechanisms of activation by ERBB2 extracellular domain (ECD) variants are not fully understood, we aimed to clarify those mechanisms and the biological functions of ERBB2 E401G. ERBB2 E401G was selected as VUS for analysis because multiple software tools predicted its pathogenicity. We prepared ERBB2 expression vectors with the E401G variant as well as vectors with S310F and E321G, which are known to be activating mutations. On the basis of wild-type ERBB2 or mutant ERBB2 expression in cell lines without ERBB2 amplification or variants, we evaluated the phosphorylation of human epidermal growth factor receptor 2 and related proteins, and investigated with molecular dynamics (MD) simulation the mechanisms conferred by the variants. The biological effects of ERBB2 E401G were also investigated, both in vitro and in vivo. We found that ERBB2 E401G enhances C-terminal phosphorylation in a way similar to S310F. MD simulation analysis revealed that these variants maintain the stability of the EGFR-HER2 heterodimer in a ligand-independent manner. Moreover, ERBB2 E401G-transduced cells showed an increased invasive capacity in vitro and an increased tumor growth capacity in vivo. Our results provide important information on the activating mechanisms of ERBB2 extracellular domain (ECD) variants and illustrate a model workflow integrating wet and dry bench processes for the analysis of VUS detected with cancer gene panel tests. The online version contains supplementary material available at 10.1007/s13402-021-00656-3.
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DOI：
10.1634/theoncologist.2009-0145
发表时间：
2010
期刊：
The oncologist
影响因子：
--
作者：
Rosen LS;Ashurst HL;Chap L
通讯作者：
Chap L
影响因子：
64.8
作者：
Hyman DM;Piha-Paul SA;Won H;Rodon J;Saura C;Shapiro GI;Juric D;Quinn DI;Moreno V;Doger B;Mayer IA;Boni V;Calvo E;Loi S;Lockhart AC;Erinjeri JP;Scaltriti M;Ulaner GA;Patel J;Tang J;Beer H;Selcuklu SD;Hanrahan AJ;Bouvier N;Melcer M;Murali R;Schram AM;Smyth LM;Jhaveri K;Li BT;Drilon A;Harding JJ;Iyer G;Taylor BS;Berger MF;Cutler RE Jr;Xu F;Butturini A;Eli LD;Mann G;Farrell C;Lalani AS;Bryce RP;Arteaga CL;Meric-Bernstam F;Baselga J;Solit DB
通讯作者：
Solit DB
影响因子：
28.4
作者：
Cobain EF;Wu YM;Vats P;Chugh R;Worden F;Smith DC;Schuetze SM;Zalupski MM;Sahai V;Alva A;Schott AF;Caram MEV;Hayes DF;Stoffel EM;Jacobs MF;Kumar-Sinha C;Cao X;Wang R;Lucas D;Ning Y;Rabban E;Bell J;Camelo-Piragua S;Udager AM;Cieslik M;Lonigro RJ;Kunju LP;Robinson DR;Talpaz M;Chinnaiyan AM
通讯作者：
Chinnaiyan AM
影响因子：
--
作者：
Hu S;Sun Y;Meng Y;Wang X;Yang W;Fu W;Guo H;Qian W;Hou S;Li B;Rao Z;Lou Z;Guo Y
通讯作者：
Guo Y
影响因子：
7
作者：
Ng, PC;Henikoff, S
通讯作者：
Henikoff, S