Kunishige Rina, Noguchi Yoshiyuki, Okamoto Naomi, Li Lei, Ono Akito, Murata Masayuki, Kano Fumi
Multimodal Cell Analysis Collaborative Research Cluster, Institute of Science Tokyo, Yokohama-shi, Kanagawa, Japan.
Cellshoot Therapeutics, Inc., Koto-ku, Tokyo, Japan.
Commun Biol. 2025 Mar 31;8(1):480. doi: 10.1038/s42003-025-07886-3.
In drug development, systematically characterizing a compound's mechanism of action (MoA), including its direct targets and effector proteins, is crucial yet challenging. Network-based approaches, unlike those focused solely on direct targets, effectively detect a wide range of cellular responses elicited by compounds. This study applied protein covariation network analysis, leveraging quantitative, morphological, and localization features from immunostained microscopic images, to elucidate the MoA of AX-53802, a novel ferroptosis inducer. From the candidate targets extracted through network analysis, GPX4 was verified as the direct target by validation experiments. Additionally, aggregates involving GPX4, TfR1, and F-actin were observed alongside iron reduction, suggesting a ferroptosis defense mechanism. Furthermore, combination therapies targeting GPX4 and FAK/Src were found to enhance cancer cell death, and MDM2, ezrin, and cortactin were identified as potential ferroptosis inhibitor targets. These findings highlight the effectiveness of network-based approaches in uncovering a compound's MoA and developing combination therapies for cancer.
在药物研发中，系统地表征化合物的作用机制（MoA），包括其直接靶点和效应蛋白，至关重要但具有挑战性。与仅关注直接靶点的方法不同，基于网络的方法能够有效地检测化合物引发的广泛细胞反应。本研究应用蛋白质共变网络分析，利用免疫染色显微镜图像的定量、形态和定位特征，以阐明新型铁死亡诱导剂AX-53802的作用机制。通过网络分析提取的候选靶点中，GPX4经验证实验确认为直接靶点。此外，观察到涉及GPX4、TfR1和F-肌动蛋白的聚集体以及铁还原现象，提示存在铁死亡防御机制。此外，发现靶向GPX4和FAK/Src的联合疗法可增强癌细胞死亡，并且MDM2、埃兹蛋白和皮层肌动蛋白被确定为潜在的铁死亡抑制剂靶点。这些发现凸显了基于网络的方法在揭示化合物作用机制和开发癌症联合疗法方面的有效性。
