Abstract
ARF GTPase protein 1 (GIT1) is a scaffold protein that is overexpressed in hepatocellular carcinoma (HCC) and colorectal cancer (CRC). GIT1 forms a complex with methionine adenosyltransferase 2B (MAT2B) that activates RAS-RAF-MEK-ERK signaling in HCC and CRC to enhance tumorigenicity. Here, we investigated in a proof-of-concept study whether a small molecule that disrupts GIT1-MAT2B interaction can be effective in HCC and CRC treatment. Since the GIT1 crystal structure is unavailable, we developed a molecular model and used computer-based drug discovery approach to screen for small molecules targeting the GIT1 ankyrin repeat domain, the region closest to where MAT2B interacts that is accessible. Of nine compounds tested, compound 3 (C3) selectively interacts with GIT1 and shows an anti-cancer effect in a GIT1-dependent manner. C3 is antiproliferative, induced apoptosis and G2/M cell cycle arrest while inhibiting colony formation and migration in liver and colon cancer cells. C3 lowered interaction between GIT1 and MAT2B, and with downstream effectors cRAF, MEK and ERK, lowering MEK activity and cyclin D1 expression. Unexpectedly, C3 stabilized GIT1 interaction with cyclin B1 while weakening cyclin B1’s interaction with components of the anaphase promoting complex, concomitant with sustained cyclin B1 expression and mitosis arrest. In mice, C3 administration was well tolerated and inhibited murine CRC growth and liver metastasis in immune competent mice and human CRC growth in the livers of nude mice. In conclusion, a small molecule inhibitor that disrupts GIT1’s normal interactome is a promising new approach to treating liver and colon cancers.
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All data generated or analysed during this study are included in this paper and in supplemental information files. All original and uncropped western blots are included in the Supplemental Material.
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Acknowledgements
Authors acknowledge Tony TW Li’s contribution in the initial screening of the GIT1 inhibitors. This work was supported by NIH grant P01CA233452 (SC Lu, ML Tomasi), Plan Nacional of I + D SAF2017-88041-R (JM Mato). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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HP, JC, WF, JW, AC, RM, YZ – conducted the experiments, data collection, drafted methods; JC, LBT, RM – prepared figures; LBT, SS, MLT, RM, JMM – critical reading of the manuscript; SCL - study concept and design, data analysis and interpretation, edited/wrote the manuscript, obtained funding, and provided overall study supervision. All authors read and approved the final manuscript.
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SCL and RM have filed a patent for small molecule inhibitors of GIT1 (Title: COMPOUNDS AND METHODS FOR TREATING CANCERS; Application No.: 63/422,672; Filed: November 4, 2022). The other authors declare that they have no competing interests.
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We confirm that all experiments were performed in accordance with relevant guidelines and regulations and approved by the Institutional Animal Care and Use Committee of Cedars-Sinai Medical Center.
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Peng, H., Chhimwal, J., Fan, W. et al. A small molecule inhibitor of ARF GTPase protein 1 limits liver and colon cancer cell growth and metastasis. Cell Death Dis (2026). https://doi.org/10.1038/s41419-026-08477-8
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DOI: https://doi.org/10.1038/s41419-026-08477-8
