一个整合的全基因组CRISPRa方法来功能化lncRNAs在药物耐药性
Assaf C Bester1, Jonathan D Lee1, Alejandro Chavez2
1Cancer Research Institute, Beth Israel Deaconess Cancer Center, Department of Medicine and Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Ludwig Center at Harvard, Harvard Medical School, Boston, MA, USA.
Cell
|April 21, 2018
概括
这项研究引入了一个新的CRISPR查平台 (DICaS),用于发现影响癌症药物耐药性的基因. 它确定了新的细胞周期,生存和信号基因,包括GAS6-AS2 lncRNA,这有助于AML的化疗耐药性.
科学领域:
- 基因组学
- 分子生物学
- 癌症研究
背景情况:
- 抗化疗是癌症死亡的主要原因.
- 赛塔拉宾是急性髓性白血病 (AML) 的关键治疗方法.
- 鉴定影响药物敏感性的遗传因素对于改善癌症治疗至关重要.
研究的目的:
- 开发和应用一个全面的全基因组CRISPR激活查平台 (DICaS),以确定癌症中细胞氨基酸敏感性的遗传决定因素.
- 在功能上表征蛋白质编码和长非编码RNA (lncRNA) 基因参与药物耐药性.
- 在AML和其他癌症中发现新的治疗点和抵抗机制.
主要方法:
- 开发一个双蛋白编码和非编码的集成CRISPR激活选平台 (DICaS).
- 使用760个人类泛癌细胞系的药物遗传学数据,初步识别可能的抗药基因.
- 使用CRISPR激活编码和lncRNA基因的全基因功能表征,包括14,701个lncRNA基因的特定CRISPR激活策略 (CaLR).
主要成果:
- DICaS成功地发现了与细胞循环调节,生存/亡以及癌症信号通路相关的新基因.
- 已发现的GAS6- AS2 lncRNA的转录激活过度激活了GAS6/ TAM通路.
- 这种过度活化代表了多种癌症,包括AML的显著抵抗机制.
结论:
- 通过分析编码和非编码遗传元素,DICaS平台是一种强大且综合的方法来发现治疗点.
- 这些发现突出了lncRNAs,如GAS6-AS2在调解化疗耐药性的作用.
- 这项研究提供了对细胞原蛋白耐药性的新见解,并提供了在AML和其他恶性瘤中克服它的潜在策略.
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