LIN28B-mediated PI3K/AKT pathway activation promotes metastasis in colorectal cancer models
Alice E. Shin, … , Peter A. Sims, Anil K. Rustgi
Alice E. Shin, … , Peter A. Sims, Anil K. Rustgi
Published January 14, 2025
Citation Information: J Clin Invest. 2025;135(8):e186035. https://doi.org/10.1172/JCI186035.
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Research Article Gastroenterology Oncology

LIN28B-mediated PI3K/AKT pathway activation promotes metastasis in colorectal cancer models

Abstract

Colorectal cancer (CRC) remains a leading cause of cancer death because of metastatic spread. LIN28B is overexpressed in 30% of CRCs and promotes metastasis, yet its mechanisms remain unclear. In this study, we genetically modified CRC cell lines to overexpress LIN28B, resulting in enhanced PI3K/AKT pathway activation and liver metastasis in mice. We developed genetically modified mouse models with constitutively active Pik3ca that form intestinal tumors progressing to liver metastases with an intact immune system, addressing the limitations of previous Pik3ca-mutant models, including long tumor latency, mixed histology, and lack of distant metastases. The PI3Kα-specific inhibitor alpelisib reduced migration and invasion in vitro and metastasis in vivo. We present a comprehensive analysis of vertical inhibition of the PI3K/AKT pathway in CRC using the FDA-approved drugs alpelisib and capivasertib (an AKT inhibitor) in combination with LY2584702 (a ribosomal protein S6 kinase inhibitor) in CRC cell lines and mouse- and patient-derived organoids. Tissue microarrays from patients with CRC verified that LIN28B and PI3K/AKT pathway activation correlate with CRC progression. These findings highlight the critical role of the LIN28B-mediated PI3K/AKT pathway in CRC metastasis, the therapeutic potential of targeted inhibition, and the promise of patient-derived organoids in precision medicine in metastatic CRC.

Authors

Alice E. Shin, Kensuke Sugiura, Secunda W. Kariuki, David A. Cohen, Samuel P. Flashner, Andres J. Klein-Szanto, Noriyuki Nishiwaki, Dechokyab De, Neil Vasan, Joel T. Gabre, Christopher J. Lengner, Peter A. Sims, Anil K. Rustgi

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Figure 1

LIN28B expression in CRC cells activates the PI3K/AKT pathway and promotes liver metastasis.

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LIN28B expression in CRC cells activates the PI3K/AKT pathway and promot...
(A) Western blot of LIN28B protein levels in LoVo and DLD-1 CRC cell lines with either EV or LIN28B overexpression vector (LIN28Bhi), normalized to GAPDH and LoVo EV (1-way ANOVA, mean ± SEM). (B) Experimental setup for in vivo colorectal liver metastasis assay. (C) Representative H&E and GFP images of liver sections from mice injected with CRC cells. Scale bars: 5 mm; scale bars for insets: 1 mm. (D) Proportion of mice that developed liver metastases (χ2 test). (E) Quantification of the size of liver metastases in each group (1-way ANOVA, mean ± SEM). (F) GSEA showing hallmark pathways enriched in LoVo LIN28Bhi cells compared with EV cells (n = 3). (G) Western blot analysis of p-AKT (Ser473) and t-AKT in CRC cells (2-tailed Student’s unpaired t test, mean ± SEM). (H) Quantification of phosphorylated protein targets involved in the PI3K/AKT pathway in LIN28Bhi cells relative to EV cells as measured by AKT pathway phosphorylation array (2-tailed Student’s unpaired t test between EV and LIN28Bhi for each protein, mean ± SEM). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.