1328:利用全基因组CRISPR-Cas9筛选鉴定胶质母细胞瘤的耐药网络

S. Budhiraja, S. Baisiwala, E. Perrault, Li Chen, Cheol Park, Chidiebere U. Awah, C. Dmello, A. Zolp, A. Sonabend, Atique U. Ahmed
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引用次数: 0

摘要

胶质母细胞瘤(GBM)是最具侵袭性和最常见的成人恶性脑肿瘤类型,每年有12,000例新诊断。即使采用目前的治疗标准——手术切除、放疗和以替莫唑胺(TMZ)为基础的化疗——中位生存期约为20个月。这部分是由于对包括TMZ在内的传统疗法的高耐药性导致复发率接近100%。是什么导致了这种耐药性的发展,这在很大程度上仍然是未知的。许多研究表明原发性和复发性肿瘤之间存在差异,但需要对耐药机制有更深入的了解。CRISPR-Cas9筛选是系统和公正的遗传分析的有力工具,我们将其用于了解TMZ抗性。我们在H4人类GBM细胞中进行了全基因组CRISPR敲除筛选,其中包含超过17,000个基因。将dmso处理的人群与tmz处理的人群进行14天的比较。在这个药敏筛选中,缺失的导体对应于tmz耐药基因,而富集的导体对应于tmz敏感基因。分析显示,已知tmz敏感基因(atg14、MSH6、MLH1和pms2)在指南中显著富集,从而验证了我们的筛选结果。然而,更重要的是,我们能够鉴定出200个与TMZ抗性有关的新基因。通路分析显示,这些基因在Hippo和Notch信号中富集,这两种信号都在化学耐药中起作用。从这个新基因列表中,我们确定了4个以前未研究过的基因。这些基因的RNA表达显著升高(p引文格式:Shreya Budhiraja, Shivani Baisiwala, Ella Perrault, Li Chen, Cheol Park, Chidiebere Awah, Crismita dello, Andrew Zolp, Adam Sonabend, Atique Ahmed)。利用全基因组CRISPR-Cas9筛选鉴定胶质母细胞瘤耐药网络[摘要]。见:美国癌症研究协会2021年年会论文集;2021年4月10日至15日和5月17日至21日。费城(PA): AACR;癌症杂志,2021;81(13 -增刊):1328。
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Abstract 1328: Using whole-genome CRISPR-Cas9 screening to identify resistance networks in glioblastoma
Glioblastoma (GBM) is the most aggressive and common type of adult malignant brain tumor, with 12,000 new diagnoses each year. Even with the current standard of care—surgical resection, radiation, and temozolomide (TMZ)-based chemotherapy—the median survival is about 20 months. This is partly due to the high rate of resistance to conventional therapy, including TMZ, leading to recurrence rates close to 100%. It remains largely unknown what drives the development of this resistance. Many studies have shown differences between primary and recurrent tumors, but a deeper understanding of resistance mechanisms is needed. CRISPR-Cas9 screening is a powerful tool for systematic and unbiased genetic analysis, which we applied to understand TMZ resistance. We performed a genome-wide CRISPR knockout screen in H4 human GBM cells, encompassing over 17,000 genes. A DMSO-treated population was compared with a TMZ-treated population over 14 days. In this drug sensitivity screen, depletion of guides corresponds to a TMZ-resistance gene, whereas enrichment of guides corresponds to a TMZ-sensitivity gene. Analysis showed that there was significant enrichment in guides for known TMZ-sensitivity genes that have been highly cited—ATG14, MSH6, MLH1, and PMS2—thus validating our screen results. However, more importantly, we were able to identify a list of 200 novel genes implicated in TMZ resistance. Pathway analysis revealed that these genes were enriched in Hippo and Notch signaling, both known to play a role in chemoresistance. From this list of novel genes, we identified 4 previously unstudied genes. These genes showed significant elevations in RNA expression (p Citation Format: Shreya Budhiraja, Shivani Baisiwala, Ella Perrault, Li Chen, Cheol Park, Chidiebere Awah, Crismita Dmello, Andrew Zolp, Adam Sonabend, Atique Ahmed. Using whole-genome CRISPR-Cas9 screening to identify resistance networks in glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1328.
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