Analysis of mTOR Gene Aberrations in Melanoma Patients and Evaluation of Their Sensitivity to PI3K-AKT-mTOR Pathway Inhibitors
Analysis of mTOR Gene Aberrations in Melanoma Patients and Evaluation of Their Sensitivity to PI3K-AKT-mTOR Pathway Inhibitors
复制标题
黑色素瘤患者 mTOR 基因畸变分析及其对 PI3K-AKT-mTOR 通路抑制剂敏感性评估
DOI：
10.1158/1078-0432.ccr-15-1110
复制
发表时间：
2016-02-15
影响因子：
11.5
通讯作者：
Guo, Jun
中科院分区：
文献类型：
--
作者：
Kong, Yan;Si, Lu;Guo, Jun
目的：mTOR是治疗癌症的有效靶点。是否可以选择携带mTOR突变的黑色素瘤患者接受PI3K-AKT-mTOR途径抑制剂的治疗还有待确定。实验设计：共纳入412例黑色素瘤标本。用Sanger测序法检测mTOR所有外显子的基因突变，并用Agilent的SureSelect靶浓缩系统进行确认。利用转录激活物样效应核酸酶技术构建稳定表达mTOR突变体的HEK293T细胞。结果：体细胞非同义突变的总发生率为10.4%(43/412)。肢端黑色素瘤(11.0%)和粘膜黑色素瘤(14.3%)中mTOR非同义突变的发生率高于慢性日光损伤(CSD；6.7%)和非CSD(3.4%)MELA-Nomas。在43例mTOR突变中，共检测到41个不同的突变，涉及25个不同的外显子。存在mTOR非同义突变的黑色素瘤患者的中位生存期明显短于无mTOR非同义突变的患者(P=0.028)。MTOR突变体在HEK293T细胞中的瞬时表达强烈激活了mTOR-p70S6K通路。在稳定表达H1968Y或P2213S mTOR突变体的HEK293T细胞中，LY294002和AZD5363在抑制PI3K-AKT-mTOR途径和细胞增殖方面比替西罗莫司或BYL719有更强的抑制作用。MTOR非同义突变可能预示着黑色素瘤较差的预后。PI3K-AKT-mTOR途径抑制剂的临床试验可能对具有特定mTOR突变的黑色素瘤患者有利。(C)2015年AACR。
Purpose: mTOR is a validated target in cancer. It remains to be determined whether melanoma patients bearing mTOR mutation could be selected for treatment with PI3K-AKT-mTOR pathway inhibitors.Experimental Design: A total of 412 melanoma samples were included. Gene aberrations in all exons of mTOR were detected by Sanger sequencing and confirmed by using Agilent's SureSelect Target Enrichment System. HEK293T cells stably expressing mTOR mutants were constructed by using transcription activator- like effector nucleases technique. Function of mTOR mutants and in vitro sensitivity of gain-of-function mTOR mutations to PI3K-AKT-mTOR pathway inhibitors were analyzed.Results: The overall incidence of somatic nonsynonymous mutations of mTOR was 10.4% (43/412). mTOR nonsynonymous mutations were relatively more frequent in acral (11.0%) and mucosal (14.3%) melanomas than in chronic sun-induced damage (CSD; 6.7%) and non-CSD (3.4%) mela-nomas. Of the 43 cases with mTOR mutations, 41 different mutations were detected, affecting 25 different exons. The median survival time for melanoma patients with mTOR nonsynonymous mutation was significantly shorter than that for patients without mTOR nonsynonymous mutation (P = 0.028). Transient expression of mTOR mutants in HEK293T cells strongly activated the mTOR-p70S6K pathway. In HEK293T cells with stable expression of H1968Y or P2213S mTOR mutants, LY294002 and AZD5363 showed higher potency than temsirolimus or BYL719 in inhibiting the PI3K-AKT-mTOR pathway and cell proliferation.Conclusions: mTOR nonsynonymous mutations are frequent in melanoma patients. mTOR nonsynonymous mutation may predict a worse prognosis of melanoma. Clinical trials with PI3K-AKT-mTOR pathway inhibitors may be beneficial for melanoma patients with specific mTOR mutations. (C) 2015 AACR.