Preclinical Assessment of a Novel CDC7 Inhibitor: Genomewide RNAi Screening Identifies Unique Synergetic and Resistance Genes,
Preclinical Assessment of a Novel CDC7 Inhibitor: Genomewide RNAi Screening Identifies Unique Synergetic and Resistance Genes,
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新型 CDC7 抑制剂的临床前评估：全基因组 RNAi 筛选鉴定出独特的协同和耐药基因，
DOI：
10.1182/blood.v118.21.3570.3570
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
M. Frattini
中科院分区：
文献类型：
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作者：
Nancy Liu;B. Bhinder;David Shum;Christina N. Ramirez;C. Radu;Hakim Djaballah;M. Frattini
摘要3570尽管进行了广泛的药物发现努力，但候选药物的失败和患者在临床上的复发仍然是持续存在的问题。虽然药物基因参与不足导致药物失败，但从头逃逸突变会导致患者复发，因此需要对基因如何影响药物反应性进行系统研究。为此，我们探索了一种基于功能性短发夹RNA（shRNA）的基因组筛选平台，旨在询问药物-基因接合并评估其对信号通路的影响。我们提出这一概念作为一种新的方式来评估候选药物的临床试验之前，使责任评估和预测临床结果。我们利用了在慢病毒颗粒中产生的阵列式shRNA文库，其特征在于几个明显的有利特征，包括一次靶向一个发夹的shRNA和动态高含量全孔显微镜成像分析。我们在不存在或存在新型CDC 7激酶抑制剂（MSK-777）的情况下以其IC 20和IC 50进行了三次平行的全基因组shRNA筛选，并鉴定了影响MSK-777敏感性和抗性的几个候选基因。这些包括增强MSK-777敏感性的增效剂和赋予MSK-777抗性的拯救剂。IPA分析将这些命中的簇映射到多个主要途径，其中包括NF-κ B途径、泛素-蛋白酶体途径、DNA复制和几个表观遗传调控基因。我们将提出并讨论这一概念以及新兴的途径，以确定关键的治疗靶点和敏感性和耐药性的生物标志物。因此，不仅允许评估调节特定药物试剂的候选基因的更广泛的适用性，而且允许鉴定定制的和更有效的治疗癌症的治疗方案。披露：没有相关的利益冲突。
Abstract 3570 Despite extensive drug discovery efforts, drug-candidate failure and patients relapsing in the clinic remain as persistent problems. While insufficient drug-gene engagement leads to drug failure, de novo escape mutations give rise to patients relapsing, calling the need for systemic studies on how genes influence drug responsiveness. Towards this end, we have explored a functional short hairpin RNA (shRNA) based genomic screening platform aimed at interrogating drug-gene engagement and assessing its consequences on signaling pathways. We propose this concept as a novel way to evaluate drug candidates prior to clinical trials enabling liability assessment and predicting clinical outcome. We took advantage of the arrayed shRNA library produced in lentiviral particles and characterized by several obvious advantageous features including shRNA targeting one hairpin at a time and on the fly high content whole well microscopy imaging analysis. We carried out three parallel genomewide shRNA screens in the absence or presence of the novel CDC7 kinase inhibitor (MSK-777) at its IC20 and IC50 and have identified several gene candidates that influence MSK-777 sensitivity and resistance. These include synergizers that enhance MSK-777 sensitivity and rescuers that confer MSK-777 resistance. IPA analysis mapped clusters of these hits to multiple major pathways among them were the NF-kB pathway, the ubiquitin-proteasome pathway, DNA replication, and several epigenetic regulatory genes. We will present and discuss this concept together with the emerging pathways as a means to identify both key therapeutic targets and biomarkers of sensitivity and resistance. Thus, allowing for not only a broader applicability of assessing candidate genes that modulate specific drug agents, but also for the identification of a tailored and more efficacious therapeutic regimen to treat cancer. Disclosures: No relevant conflicts of interest to declare.