Xue Jian, Li Meng, Kang Li, Wang Meiting, Yin Jiabin, Sun Donghui, Deng Yaqi, Wei Qinghua, Wong Jiemin, Zhu Tong, Liu Shunying
Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai, 200062, China.
Shanghai Key Laboratory of Regulatory Biology, Fengxian District Central Hospital-ECNU Joint Center of Translational Medicine, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, 200241, China.
Cell Death Dis. 2025 Jul 19;16(1):537. doi: 10.1038/s41419-025-07765-z.
Osteosarcoma (OS) has been defined as one of the most intricate and formidable malignant bone tumors, and there has been no significant improvement in targeted therapies for OS over the past 50 years. Therefore, it is crucial to identify new potential drug targets for OS. Here, we have developed a label-free activity-based protein profiling (ABPP) using a stereochemically selective probe from an in-house patrimonial library of covalent small molecule compounds to identify an anti-OS target. Phenotypic screening resulted in the discovery of a selective inhibitor (S,R)-4v that potently suppresses the proliferation of OS 143B cells with an IC value of 0.28 µM. Subsequent label-free ABPP studies identified neurite outgrowth inhibitor B (Nogo-B) as the primary cellular target for (S,R)-4v via a rapid relatively quantitative analysis using its inactive isomer as control. This finding was validated by interaction assays including pull-down, cellular thermal shift assay (CETSA), molecular docking and functional studies. Mechanistic investigations revealed that the apoptotic effect induced by (S,R)-4v was mediated through Nogo-B inhibition of the PI3K/AKT-dependent NF-κB pathway. Altogether, this study presents a novel strategy that couples anti-OS compound screening with target identification and successfully identifies Nogo-B as a potential candidate for targeted OS therapy.
骨肉瘤（OS）被定义为最复杂且可怕的恶性骨肿瘤之一，在过去50年里，OS的靶向治疗没有显著进展。因此，确定OS新的潜在药物靶点至关重要。在此，我们利用来自内部共价小分子化合物祖传文库的立体化学选择性探针，开发了一种无标记的基于活性的蛋白质谱分析（ABPP）方法，以鉴定抗OS靶点。表型筛选发现了一种选择性抑制剂（S，R）-4v，它能有效抑制OS 143B细胞的增殖，IC值为0.28µM。随后的无标记ABPP研究通过使用其无活性异构体作为对照的快速相对定量分析，确定神经突生长抑制因子B（Nogo-B）是（S，R）-4v的主要细胞靶点。通过包括下拉、细胞热位移分析（CETSA）、分子对接和功能研究在内的相互作用分析验证了这一发现。机制研究表明，（S，R）-4v诱导的凋亡效应是通过Nogo-B抑制PI3K/AKT依赖的NF-κB途径介导的。总之，本研究提出了一种将抗OS化合物筛选与靶点鉴定相结合的新策略，并成功将Nogo-B鉴定为OS靶向治疗的潜在候选靶点。
