Full
Outline
To screen novel molecular glues targeting ovarian tumor domain deubiquitinase with linear linkage specificity (OTULIN) and provide a new therapeutic strategy for gastric cancer.
Gastric cancer cell lines with high OTULIN expression (AGS-NC, SNU601-NC) and OTULIN-knockout cells (AGS-sgOTULIN, SNU601-sgOTULIN) were used. Experimental groups included: control (DMSO), LTL-04-061 concentration gradients (0.1, 0.3, 1.0, 3.0, 5.0, 7.0, 10.0 μmol/L), and proteasome inhibitor intervention (MG-132/MLN4924+5 μmol/L LTL-04-061). OTULIN protein expression was analyzed by Western blotting, while mRNA levels were quantified by qPCR. Protein interactions were validated by co-immunoprecipitation (Co-IP). Molecular docking and 100 ns molecular dynamics (MD) simulations were performed to analyze complex conformations. Anti-proliferative effects were assessed using CCK-8, cell proliferation, and colony formation assays.
LTL-04-061 degraded OTULIN protein in a concentration-dependent manner (0.1 to 10.0 μmol/L), with significant degradation at 5 μmol/L. At this concentration, OTULIN mRNA levels remained unchanged (P>0.05). Degradation was blocked by MG-132/MLN4924, indicating dependence on the CRBN-mediated ubiquitin-proteasome pathway. Co-IP confirmed LTL-04-061-induced OTULIN-CRBN ternary complex formation. Key interactions involved OTULIN HIS339 and CRBN TRP378/HIS380. MD simulations showed stable complex conformation after 50 ns (RMSD fluctuation <0.1 nm), sustained hydrogen bonds (80 to 100 ns), and stable radius of gyration (Rg) and solvent-accessible surface area (SASA), indicating no structural expansion/collapse. LTL-04-061 concentration-dependently suppressed gastric cancer cell proliferation and colony formation. CCK-8 assays revealed IC50 values of 8.95 μmol/L (AGS) and 11.70 μmol/L (SNU601). OTULIN knockout significantly attenuated this inhibition (P<0.01).
OTULIN exerts pro-oncogenic effects in gastric cancer. The molecular glue LTL-04-061 targets OTULIN for CRBN-dependent ubiquitin-proteasomal degradation, thereby suppressing malignant proliferation. This identifies LTL-04-061 as a promising precursor for molecularly targeted gastric cancer therapy.
91
39
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
关闭全屏
No. 86 Xueyuan South Road, Haidian District, Beijing
010-62662699(Operation and Technical Support)
qkjq@cast.org.cn
Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House
