Revealing Temozolomide Resistance Mechanisms via Genome-Wide CRISPR Libraries.
Revealing Temozolomide Resistance Mechanisms via Genome-Wide CRISPR Libraries.
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通过全基因组CRISPR文库揭示替莫唑胺耐药机制。
DOI：
10.3390/cells9122573
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发表时间：
2020-12-01
期刊：
影响因子：
6
通讯作者：
Menck CFM
中科院分区：
文献类型：
--
作者：
Rocha CRR;Reily Rocha A;Molina Silva M;Rodrigues Gomes L;Teatin Latancia M;Andrade Tomaz M;de Souza I;Karolynne Seregni Monteiro L;Menck CFM
胶质母细胞瘤是一种严重的脑肿瘤，预后差，治疗选择少。替莫唑胺（TMZ）是这些选择之一，然而，成功有限，失败主要是由于肿瘤耐药性。在这项工作中，在人类胶质母细胞瘤细胞系中转导了用于基因敲除或激活的全基因组CRISPR-Cas9慢病毒筛选文库，旨在鉴定调节TMZ抗性的基因。通过下一代测序（NGS）鉴定TMZ处理后存活细胞中两个文库中富集的sgRNA。对敲除筛选的候选基因的途径分析揭示了几种富集的途径，包括错配修复和Sonic Hedgehog途径。沉默排名前10位的三个基因（MSH 2，PTCH 2和CLCA 2）证实了TMZ诱导的细胞死亡保护。此外，CRISPR激活文库显示NRF 2和Wnt途径参与TMZ抗性。一致地，FZD 6、CTNNB 1或NRF 2基因的过表达显著增加TMZ处理后的细胞存活。此外，在该筛选中检测到的NRF 2和相关基因与胶质母细胞瘤患者的生存率呈显著负相关。最后，来自敲除或激活筛选的几个候选基因可被抑制剂或小分子靶向，其中一些已经用于临床。
Glioblastoma is a severe type of brain tumor with a poor prognosis and few therapy options. Temozolomide (TMZ) is one of these options, however, with limited success, and failure is mainly due to tumor resistance. In this work, genome-wide CRISPR-Cas9 lentiviral screen libraries for gene knockout or activation were transduced in the human glioblastoma cell line, aiming to identify genes that modulate TMZ resistance. The sgRNAs enriched in both libraries in surviving cells after TMZ treatment were identified by next-generation sequencing (NGS). Pathway analyses of gene candidates on knockout screening revealed several enriched pathways, including the mismatch repair and the Sonic Hedgehog pathways. Silencing three genes ranked on the top 10 list (MSH2, PTCH2, and CLCA2) confirm cell protection from TMZ-induced death. In addition, a CRISPR activation library revealed that NRF2 and Wnt pathways are involved in TMZ resistance. Consistently, overexpression of FZD6, CTNNB1, or NRF2 genes significantly increased cell survival upon TMZ treatment. Moreover, NRF2 and related genes detected in this screen presented a robust negative correlation with glioblastoma patient survival rates. Finally, several gene candidates from knockout or activation screening are targetable by inhibitors or small molecules, and some of them have already been used in the clinic.
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影响因子：
64.8
作者：
Konermann S;Brigham MD;Trevino AE;Joung J;Abudayyeh OO;Barcena C;Hsu PD;Habib N;Gootenberg JS;Nishimasu H;Nureki O;Zhang F
通讯作者：
Zhang F
影响因子：
--
作者：
da Silva PBG;Teixeira Dos Santos MC;Rodini CO;Kaid C;Pereira MCL;Furukawa G;da Cruz DSG;Goldfeder MB;Rocha CRR;Rosenberg C;Okamoto OK
通讯作者：
Okamoto OK
DOI：
10.1038/nrd.2016.238
发表时间：
2017-02
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
Fellmann C;Gowen BG;Lin PC;Doudna JA;Corn JE
通讯作者：
Corn JE
全基因组 CRISPR-Cas9 筛选鉴定出 NF-kappa B/E2F6 负责胶质母细胞瘤中与 EGFRvIII 相关的替莫唑胺耐药性
影响因子：
15.1
作者：
Huang, Kai;Liu, Xing;Kang, Chunsheng
通讯作者：
Kang, Chunsheng
影响因子：
10.5
作者：
Jaramillo MC;Zhang DD
通讯作者：
Zhang DD