CK2 和 ATM 激酶的协同阻断通过 ROS 过量产生驱动 VHL 缺陷型肾癌细胞凋亡

发表日期：2022-03-21

Cooperative Blockade of CK2 and ATM Kinases Drives Apoptosis in VHL-Deficient Renal Carcinoma Cells through ROS Overproduction

Renal cell carcinoma (RCC) is the eighth leading malignancy in the world, accounting for 4% of all cancers with poor outcome when metastatic. Protein kinases are highly druggable proteins, which are often aberrantly activated in cancers. The aim of our study was to identify candidate targets for metastatic clear cell renal cell carcinoma therapy, using chemo-genomic-based high-throughput screening. We found that the combined inhibition of the CK2 and ATM kinases in renal tumor cells and patient-derived tumor samples induces synthetic lethality. Mechanistic investigations unveil that this drug combination triggers apoptosis through HIF-2α-(Hypoxic inducible factor HIF-2α) dependent reactive oxygen species (ROS) overproduction, giving a new option for patient care in metastatic RCC.

肾细胞癌 (RCC) 是世界上第八大恶性肿瘤，占所有转移性预后不良癌症的 4%。蛋白激酶是高度可药用的蛋白质，通常在癌症中被异常激活。我们研究的目的是使用基于化学基因组的高通量筛选来确定转移性透明细胞肾细胞癌治疗的候选靶点。我们发现在肾肿瘤细胞和患者来源的肿瘤样本中联合抑制 CK2 和 ATM 激酶可诱导合成致死率。机制研究表明，这种药物组合通过 HIF-2α-（缺氧诱导因子 HIF-2α）依赖性活性氧（ROS）过量产生触发细胞凋亡，为转移性肾细胞癌的患者护理提供了新的选择。

T75细胞培养瓶

Kinase-targeted agents demonstrate antitumor activity in advanced metastatic clear cell renal cell carcinoma (ccRCC), which remains largely incurable. Integration of genomic approaches through small-molecules and genetically based high-throughput screening holds the promise of improved discovery of candidate targets for cancer therapy. The 786-O cell line represents a model for most ccRCC that have a loss of functional pVHL (von Hippel-Lindau). A multiplexed assay was used to study the cellular fitness of a panel of engineered ccRCC isogenic 786-O VHL− cell lines in response to a collection of targeted cancer therapeutics including kinase inhibitors, allowing the interrogation of over 2880 drug–gene pairs. Among diverse patterns of drug sensitivities, investigation of the mechanistic effect of one selected drug combination on tumor spheroids and ex vivo renal tumor slice cultures showed that VHL-defective ccRCC cells were more vulnerable to the combined inhibition of the CK2 and ATM kinases than wild-type VHL cells. Importantly, we found that HIF-2α acts as a key mediator that potentiates the response to combined CK2/ATM inhibition by triggering ROS-dependent apoptosis. Importantly, our findings reveal a selective killing of VHL-deficient renal carcinoma cells and provide a rationale for a mechanism-based use of combined CK2/ATM inhibitors for improved patient care in metastatic VHL-ccRCC.

靶向激酶的药物在晚期转移性透明细胞肾细胞癌 (ccRCC) 中表现出抗肿瘤活性，而这在很大程度上仍然无法治愈。通过小分子和基于遗传的高通量筛选整合基因组方法有望改进癌症治疗候选靶点的发现。786-O 细胞系代表了大多数功能性 pVHL 缺失的 ccRCC 模型 (von Hippel-Lindau)。使用多重测定来研究一组工程化 ccRCC 同基因 786-O VHL 的细胞适应性-细胞系响应包括激酶抑制剂在内的一系列靶向癌症治疗，允许对超过 2880 种药物-基因对进行研究。在多种药物敏感性模式中，对一种选定药物组合对肿瘤球体和离体肾肿瘤切片培养物的机制影响的研究表明，与野生型相比，VHL 缺陷型 ccRCC 细胞更容易受到 CK2 和 ATM 激酶联合抑制的影响。型 VHL 细胞。重要的是，我们发现 HIF-2α 作为一种关键介质，通过触发 ROS 依赖性细胞凋亡来增强对联合 CK2/ATM 抑制的反应。重要的是，我们的研究结果揭示了 VHL 缺陷型肾癌细胞的选择性杀伤，并为基于机制的 CK2/ATM 抑制剂联合使用以改善转移性 VHL-ccRCC 的患者护理提供了依据。

细胞工厂

Genomic analyses have revealed relationships between synthetic lethal interactions and genetic lesions that may be exploited therapeutically [21]. Chemo-genetic screens revealed CK2 and ATM kinases as synthetic lethal targets in VHL-deficient renal carcinoma cells. This vulnerability was characterized using various models, including cells grown as monolayers, tumor spheroid cultures and tumor tissue slices from xenografts and clinical patient samples. Combined inhibition of CK2 and ATM eliminated VHL-deficient renal carcinoma cells by increasing NOX4-mediated ROS production. Importantly, HIF-2α acts as a key determinant that potentiates this response. Overall, given the demonstrated safety of CX-4945 in initial human studies, these preclinical data may justify the implementation of clinical trials using CX-4945 in combination with ATM inhibitors in a subset of HIF-2α-expressing VHL-defective ccRCC patients. Our findings have translational implications which are currently evaluated in the frame of the preclinical trial NCT03571438 .

基因组分析揭示了合成致死相互作用与可用于治疗的遗传损伤之间的关系 [ 21]。化学遗传学筛选显示 CK2 和 ATM 激酶是 VHL 缺陷型肾癌细胞中的合成致死靶标。使用各种模型来表征这种脆弱性，包括生长为单层的细胞、肿瘤球体培养物和来自异种移植物和临床患者样本的肿瘤组织切片。CK2 和 ATM 的联合抑制通过增加 NOX4 介导的 ROS 产生来消除 VHL 缺陷型肾癌细胞。重要的是，HIF-2α 是增强这种反应的关键决定因素。总体而言，鉴于 CX-4945 在最初的人体研究中被证明是安全的，这些临床前数据可能证明使用 CX-4945 与 ATM 抑制剂联合在表达 HIF-2α 的 VHL 缺陷型 ccRCC 患者的子集中实施临床试验是合理的。