PO.CH01.07 · 化学
从表型筛选到机制洞察：胶质母细胞瘤化疗增敏剂的理性发现
From phenotypic screening to mechanistic insight: Rational discovery of chemosensitizers for glioblastoma
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
多形性胶质母细胞瘤（GBM）是最具侵袭性的原发性脑肿瘤，其特征为进展迅速、治疗耐药以及几乎普遍的复发。尽管采取了多模式治疗策略——包括最大范围手术切除，随后进行放疗和基于替莫唑胺（TMZ）的化疗——中位生存期仍然惨淡，大多数患者在确诊后一年内死亡。TMZ和电离辐射主要通过诱导DNA损伤（包括单链和双链断裂）发挥其细胞毒性作用。细胞DNA损伤应答（DDR）涵盖由激酶、修复酶和检查点调节因子构成的复杂信号网络，协调对这些损伤的检测和修复。DDR通路（包括由ATR/ATM信号、PARP依赖性修复以及细胞周期检查点（如WEE1、CDK1）介导的通路）的失调或过度激活，被认为与GBM显著的治疗耐药有关。在本研究中，我们致力于理性设计和合成新型小分子，旨在增强TMZ和放射诱导的GBM细胞毒性。采用基于结构的设计原则，跨五个聚焦系列生成了七十余种化合物。在TMZ耐药的T98G细胞系和患者来源的GBM培养物中进行表型筛选，以评估内在细胞毒性和TMZ增敏作用。若干先导候选物相比临床DDR抑制剂表现出更优的化疗增敏效力。有趣的是，广泛的激酶谱分析显示不存在经典的DDR激酶抑制，提示存在潜在的非经典作用机制。为阐明该机制，进行了全面的定量蛋白质组学分析，揭示了与DNA修复、应激反应和细胞周期控制相关通路的调节。最有前景的候选物表现出良好的药代动力学和毒理学特性，以及在体内有效的血脑屏障穿透能力。
本工作得到捷克共和国卫生部拨款（NW24J-03-00005）以及军事医学院长期发展计划——大规模杀伤性武器II医疗保健挑战（DZRO-VLF22-ZHN II）的支持。
查看英文原文 English abstract
Glioblastoma multiforme (GBM) represents the most aggressive form of primary brain tumor, characterized by rapid progression, therapeutic resistance, and near-universal recurrence. Despite multimodal treatment strategies-comprising maximal surgical resection followed by radiotherapy and temozolomide (TMZ)-based chemotherapy-median survival remains dismal, with most patients succumbing within one-year post-diagnosis. TMZ and ionizing radiation exert their cytotoxic effects primarily through the induction of DNA lesions, including single- and double-strand breaks. The cellular DNA damage response (DDR), encompassing a complex signaling network of kinases, repair enzymes, and checkpoint regulators, orchestrates the detection and repair of these lesions. Dysregulation or hyperactivation of DDR pathways-including those mediated by ATR/ATM signaling, PARP-dependent repair, and cell cycle checkpoints (e.g., WEE1, CDK1)-has been implicated in the pronounced therapeutic resistance of GBM. In this study, we pursued the rational design and synthesis of novel small molecules aimed at potentiating TMZ- and radiation-induced cytotoxicity in GBM. Over seventy compounds were generated across five focused series using structure-based design principles. Phenotypic screening was conducted in the TMZ-resistant T98G line and patient-derived GBM cultures to evaluate both intrinsic cytotoxicity and TMZ sensitization. Several lead candidates demonstrated superior chemosensitization efficacy compared to clinical DDR inhibitors. Interestingly, broad kinase profiling revealed an absence of canonical DDR kinase inhibition, indicating a potential non-classical mechanism of action. To delineate this mechanism, comprehensive quantitative proteomic analyses were performed, revealing modulation of pathways associated with DNA repair, stress response, and cell cycle control. The most promising candidates exhibited favorable pharmacokinetic and toxicological properties, along with effective blood-brain barrier penetration in vivo.
This work has been supported by the grant from the Ministry of Health of the Czech Republic (NW24J-03-00005); and by the Long-term development plan Military Faculty of Medicine, Healthcare Challenges of WMD II (DZRO-VLF22-ZHN II).
利益披露 Disclosure
L. Gorecki, None..
L. Muckova, None..
M. Sadibolova, None..
O. Soukup, None.