PO.ET02.04 · 实验与分子治疗
通过双载荷创新克服异质性和耐药性以推进双载荷ADC发现项目
Enhancing dual-payload ADC discovery projects by overcoming heterogeneity and resistance through dual-payload innovation
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
双载荷抗体药物偶联物（ADC）已成为一种创新且有前景的下一代ADC形式，通过递送两种细胞毒剂以经由协同效应增强疗效、减轻耐药，同时不伴随毒性风险的增加，旨在应对应答不佳的癌症和患者快速复发这一挑战。双载荷ADC的成功取决于合适的双载荷组合。DNA损伤应答（DDR）抑制剂正日益被评估作为ADC的载荷，其中拓扑异构酶1（Top 1）抑制剂是最常用的一类。Top 1抑制在复制叉处捕获DNA断裂，同时阻断DDR通路切断了修复逃逸途径，导致复制灾难和选择性合成致死性细胞死亡。为识别一种能够克服结直肠癌（CRC）肿瘤异质性和传统耐药的合理设计的双载荷组合，对Top 1抑制剂与一组具有不同机制的DDR抑制剂的协同配对进行了筛选和评估。首先在CRC细胞组合中研究了单载荷活性，随后在部分选定的CRC细胞系和ADC相关耐药细胞系中评估了双载荷组合协同作用。在2D和3D细胞毒性评估模型中，一对Top 1抑制剂+CHK抑制剂的组合在CRC细胞系和ADC相关耐药细胞系中显示出显著的强效协同作用。通过细胞周期和凋亡研究、生物标志物的WB以及RNA测序进一步探索了该组合的机制研究，揭示了这种协同作用源于协调的细胞周期阻滞和增强的凋亡诱导，并得到DDR通路转录组分析的支持。关键的是，该双载荷组合在一组健康人细胞系中未显示毒性增加，在体外CD34+ HSPC髓系祖细胞增殖和分化测定中显示有限或相加的血液毒性，表明治疗窗口可能得到改善。这些研究策略和发现加速了双载荷ADC的创新研究，尤其是在双载荷组合作为克服癌症异质性和耐药性的有前景策略的概念验证阶段。
查看英文原文 English abstract
Dual-payload antibody-drug conjugates (ADCs) have emerged as an innovative promising next-generation ADC format, delivering two cytotoxic agents to enhance efficacy through synergistic effects, mitigate resistance, without an accompanying increase in toxicity risk, aiming to address the challenge of poor-responsive cancers and rapid patient relapses. The success of dual-payload ADCs hinges on the appropriate dual-payload combination.DNA-damage response (DDR) inhibitors are increasingly being evaluated as payloads for ADCs, with Topoisomerase 1 (Top 1) inhibitors being the most commonly used class. Top 1 inhibition traps DNA breaks at replication forks, simultaneously blocking the DDR pathway severs the repair escape route, leading to replication catastrophe and selective synthetic-lethal cell death.To identify a rationally designed dual-payload combination that overcomes tumor heterogeneity and conventional resistance of colorectal cancer (CRC), synergistic pairs of Top 1 inhibitors and a group of DDR inhibitors with different mechanisms were screened and evaluated. Single payload activities were first investigated in CRC cell panel, and dual-payload combination synergies were then assessed in some selected CRC cell lines and ADC-related drug-resistant cell lines. A pair of Top 1 inhibitor + CHK inhibitor combination showed significant potent synergy in CRC cell lines and ADC-related drug-resistant cell lines with 2D and 3D cytotoxicity evaluation models. Mechanistic studies of this combination were further explored with cell cycle and apoptosis studies, WB of biomarkers, as well as RNA-Sequencing, revealing that the synergy stemmed from coordinated cell cycle arrest and enhanced apoptosis induction, supported by transcriptomic profiling of DDR pathways. Critically, the dual-payload combination showed no increased toxicity in a group of healthy human cell lines and limited or additive hematotoxicity in the in vitro CD34+ HSPCs myeloid progenitor cell proliferation and differentiation assays, indicating a potentially improved therapeutic window. These study strategies and findings accelerate dual-payload ADC innovative research, especially in the stage of proof of concept for dual-payload combination as a promising strategy to overcome cancer heterogeneity and resistance
利益披露 Disclosure
Y. Meng, None..
L. Chai, None..
W. Liu, None..
X. Yang, None..
M. Kang, None..
R. Song, None..
Z. Li, None..
Y. Zhang, None..
Z. Bao, None..
Y. Bi, None..
Y. Zhao, None..
W. Fan, None..
X. Li, None..
L. Li, None..
T. Bing, None.