Wang Weishu, Wang Hao, Zhang Zeyu, Liu Xin, Hu Binbin, Tian Feng, Ye Zhou, Shi Linqi, Yu Zhilin
Key Laboratory of Functional Polymer Materials, Ministry of Education, State Key Laboratory of Medicinal Chemical Biology, Institute of Polymer Chemistry, College of Chemistry, Nankai University, 94 Weijin Road, Tianjin, 300071, China.
Applied Oral Sciences and Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong S.A.R., 999077, China.
Adv Mater. 2025 Jul;37(28):e2420399. doi: 10.1002/adma.202420399. Epub 2025 May 7.
Liquid-liquid phase separation (LLPS) of proteins and nucleic acids into membraneless organelles (MLOs) plays a critical role in sustaining fundamental physiological processes. However, creating artificial coacervate droplets in living cells from exogenous molecules and modulating the functions of MLOs remain challenging. To address this concern, here we reported enzyme-induced in situ phase separation of peptides into droplets targeting MLO stress granule (SG) for enhanced cancer chemotherapy. The peptide YF containing two sulfated tyrosine residues undergoes sulfatase-responsive LLPS into droplets. Cellular studies confirm in situ phase separation of YF selectively in sulfatase-overexpressing cancer cells. By integrating with appropriate ligands, the in situ-formed droplets d-YF-L coacervate with SGs driven by association between the ligand with SG key component protein G3BP2. Mechanistic studies illustrate that the in situ-formed droplets enhance the cytotoxicity of sorafenib via activating caspase-dependent apoptosis. Furthermore, animal experiments confirm that administration of the in situ-formed droplets with sorafenib significantly inhibits tumor growth in murine models bearing tumors, accompanied by an excellent biosafety profile. The findings in this study elucidate an innovative approach for in situ formulation of coacervate droplets within tumor cells and a new material for targeting membraneless organelles, thus providing a promising new strategy for disease organelle-targeted therapy in the future.
蛋白质和核酸的液-液相分离（LLPS）形成无膜细胞器（MLOs）在维持基本生理过程中起着关键作用。然而，利用外源性分子在活细胞中创建人工凝聚液滴并调节MLOs的功能仍然具有挑战性。为了解决这一问题，我们在此报告了酶诱导的肽原位相分离形成靶向MLO应激颗粒（SG）的液滴，以增强癌症化疗效果。含有两个硫酸化酪氨酸残基的肽YF在硫酸酯酶作用下发生响应性LLPS形成液滴。细胞研究证实YF在硫酸酯酶过表达的癌细胞中选择性地发生原位相分离。通过与适当的配体整合，原位形成的液滴d-YF-L与SGs凝聚，这是由配体与SG关键组成蛋白G3BP2之间的结合驱动的。机制研究表明，原位形成的液滴通过激活半胱天冬酶依赖性凋亡增强了索拉非尼的细胞毒性。此外，动物实验证实，将原位形成的液滴与索拉非尼联合给药可显著抑制荷瘤小鼠模型中的肿瘤生长，且具有良好的生物安全性。本研究结果阐明了一种在肿瘤细胞内原位制备凝聚液滴的创新方法以及一种靶向无膜细胞器的新材料，从而为未来疾病细胞器靶向治疗提供了一种有前景的新策略。
