Functional Genomic Identification of Predictors of Sensitivity and Mechanisms of Resistance to Multivalent Second-Generation TRAIL-R2 Agonists.
Functional Genomic Identification of Predictors of Sensitivity and Mechanisms of Resistance to Multivalent Second-Generation TRAIL-R2 Agonists.
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DOI：
10.1158/1535-7163.mct-21-0532
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发表时间：
2022-04-01
影响因子：
5.7
通讯作者：
McDade SS
中科院分区：
文献类型：
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作者：
Grinkevitch V;Wappett M;Crawford N;Price S;Lees A;McCann C;McAllister K;Prehn J;Young J;Bateson J;Gallagher L;Michaut M;Iyer V;Chatzipli A;Barthorpe S;Ciznadija D;Sloma I;Wesa A;Tice DA;Wessels L;Garnett M;Longley DB;McDermott U;McDade SS
多价第二代 TRAIL-R2 激动剂目前正处于临床前开发后期和早期临床试验阶段。在此，我们使用具有代表性的第二代药物 MEDI3039 来解决这些药物面临的两个主要临床挑战：缺乏预测性生物标志物来支持患者选择和耐药性的出现。全基因组 CRISPR 敲除筛选值得注意的是，在 TP53 野生型模型中，除了经典 TRAIL-R2 途径（caspase-8、FADD、BID）以及 p53 和 BAX 之外，缺乏耐药机制，而 CRISPR 激活筛选发现细胞死亡抑制剂 MCL-1 和 BCL-XL 是抑制 MEDI3039 诱导的细胞死亡的机制。高通量药物筛选未能识别与 MEDI3039 反应相关的基因组改变；然而，转录组学分析揭示了 MEDI3039 敏感性与外源性凋亡途径核心成分的表达之间存在显着关联，最显着的是实体瘤细胞系中其主要凋亡效应器 caspase-8。对结直肠细胞系和患者来源的异种移植物的进一步分析确定了 caspase-8 与其内源性调节因子 FLIP(L) 的表达比率可预测几种主要实体瘤类型和由 caspase-8:MCL-1 比率指示的进一步子集对 MEDI3039 的敏感性。随后 MEDI3039 对 TRAIL-R2、caspase-8、FADD 和 BID 敲除模型的 60 种具有不同作用机制的化合物进行组合筛选，鉴定出两种凋亡蛋白抑制剂 (IAP)，它们与 MEDI3039 表现出强大的协同作用，只能逆转 BID 删除模型中的耐药性。总之，我们将 caspase-8:FLIP(L) 和 caspase-8:MCL-1 的比率确定为第二代 TRAIL-R2 激动剂的潜在预测生物标志物，并将 FADD 和 caspase-8 等关键效应子的丧失确定为实体瘤临床耐药性的可能驱动因素。
Multivalent second-generation TRAIL-R2 agonists are currently in late preclinical development and early clinical trials. Herein, we use a representative second-generation agent, MEDI3039, to address two major clinical challenges facing these agents: lack of predictive biomarkers to enable patient selection and emergence of resistance. Genome-wide CRISPR knockout screens were notable for the lack of resistance mechanisms beyond the canonical TRAIL-R2 pathway (caspase-8, FADD, BID) as well as p53 and BAX in TP53 wild-type models, whereas a CRISPR activatory screen identified cell death inhibitors MCL-1 and BCL-XL as mechanisms to suppress MEDI3039-induced cell death. High-throughput drug screening failed to identify genomic alterations associated with response to MEDI3039; however, transcriptomics analysis revealed striking association between MEDI3039 sensitivity and expression of core components of the extrinsic apoptotic pathway, most notably its main apoptotic effector caspase-8 in solid tumor cell lines. Further analyses of colorectal cell lines and patient-derived xenografts identified caspase-8 expression ratio to its endogenous regulator FLIP(L) as predictive of sensitivity to MEDI3039 in several major solid tumor types and a further subset indicated by caspase-8:MCL-1 ratio. Subsequent MEDI3039 combination screening of TRAIL-R2, caspase-8, FADD, and BID knockout models with 60 compounds with varying mechanisms of action identified two inhibitor of apoptosis proteins (IAP) that exhibited strong synergy with MEDI3039 that could reverse resistance only in BID-deleted models. In summary, we identify the ratios of caspase-8:FLIP(L) and caspase-8:MCL-1 as potential predictive biomarkers for second-generation TRAIL-R2 agonists and loss of key effectors such as FADD and caspase-8 as likely drivers of clinical resistance in solid tumors.