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Ran Wang, Yatong Yuan, Huarong Shao, Yuehao Sun, Changcheng Lai, Mengrui Zhang, Wenjing Song, Tao Zhang, Fengfeng Zhuang, Qixin Chen, Peixue Ling, Xintian Shao. Real-Time Visualization of Drug-Target Interactions in Native Subcellular Microenvironments for Lysosome-targeted Drug Discovery[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2025.101428
Citation: Ran Wang, Yatong Yuan, Huarong Shao, Yuehao Sun, Changcheng Lai, Mengrui Zhang, Wenjing Song, Tao Zhang, Fengfeng Zhuang, Qixin Chen, Peixue Ling, Xintian Shao. Real-Time Visualization of Drug-Target Interactions in Native Subcellular Microenvironments for Lysosome-targeted Drug Discovery[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2025.101428

Real-Time Visualization of Drug-Target Interactions in Native Subcellular Microenvironments for Lysosome-targeted Drug Discovery

doi: 10.1016/j.jpha.2025.101428
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This work is supported by the Key Technologies Research and Development Program, China (Grant No.: 2021YFC2103100), Beijing Municipal Natural Science FoundationKey Research Project of the Daxing, China (Grant No.: L246029), National Natural Science Foundation of China (Grant Nos.: 82302743 and 22107059), Key Technology and Development Program of Shandong Province, China (Project No.: 2022SFGC0103), Young Elite Scientists Sponsorship Program by China Association for Science and Technology, China (Project No.: CACM-2023-QNRC1-02), Natural Science Foundation of Shandong Province, China (Grant Nos.:ZR2021QH057 and ZR2022QH304), National Administration of Traditional Chinese Medicine-Shandong Province Joint Construction of Traditional Chinese Medicine Technology Project, China (Project No.: GZY-KJS-SD-2023-085), Taishan Scholars Program, China (Project Nos.: TSQN202211221 and TSQN202408252), Shandong Science Fund for Excellent Young Scholars, China (Grant No.: ZR2022YQ66), Shandong Province Traditional Chinese Medicine Science and Technology Project, China (Project No.: Q- 2023059), Shandong Province Medical and Health Science and Technology Project, China (Project No.: 202404070376).

  • Received Date: Apr. 29, 2025
  • Accepted Date: Jul. 31, 2025
  • Rev Recd Date: Jul. 24, 2025
  • Available Online: Aug. 02, 2025
  • Conventional ex vivo drug screening platforms struggle to recapitulate native subcellular microenvironments, leading to high off-target rates and compromised discovery of bioactive compounds. To address this, we developed subcellular target- tracking fluorescent-visualization-based interaction screening (SubTrack-FVIS), a platform combining super-resolution imaging with target-specific fluorescent tagging. SubTrack-FVIS first maps nanoscale spatial distributions of drug targets within living cells, then screens compound libraries to identify molecules specifically binding to target-enriched domains, and finally quantifies drug-target interactions through super- resolution imaging tracking. Compared to traditional toolbox, SubTrack-FVIS reduces off-target effects by evaluating compound binding within native subcellular architectures. When applied to the lysosomal vacuolar H+-ATPases (V-ATPase) subunit, ATP6V1A, a validated anti-cancer target, this approach identified for lysosomal alkalization fluorescent drug (LAFD) as a potent inhibitor. Super-resolution imaging revealed LAFD's dynamic binding to ATP6V1A clusters, enabling real-time visualization of V-ATPase inhibition and subsequent lysosomal destabilization. Crucially, SubTrack-FVIS uncovered LAFD's unique mechanism of blocking autophagosome-lysosome fusion, resolving autophagic flux obstruction at sub-100 nm resolution. This platform establishes a visualization framework for discovering drugs within physiological subcellular contexts while simultaneously decoding their mechanistic impacts, offering application potential for target-centric drug development.
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      沈阳化工大学材料科学与工程学院 沈阳 110142

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