The Anti-Tumor Effect of the Newly Developed LAT1 Inhibitor JPH203 in Colorectal Carcinoma, According to a Comprehensive Analysis

SIMPLE SUMMARY: The large neutral amino acid transporter family (LAT1-4) plays an important role in the intracellular uptake of essential amino acids. Among them, LAT1 is specifically expressed in cancer cells. A novel LAT1-specific inhibitor, JPH203, is expected to cause cancer-specific starvation and possess anti-tumor effects; however, the anti-tumor mechanism for colorectal cancer (CRC) remains unclear. In this study, we clarified the significance of LAT1 expression in cancer progression in database analysis and clinical specimens and demonstrated the utility of JPH203, in vitro and in vivo, using a unique stroma-abundant allogeneic immunoreactive mouse colorectal tumor created by the orthotopic transplantation of a mouse-derived CRC cell line and mesenchymal stem cells into mouse cecum. Comprehensive analysis with RNA sequences showed the efficacy of JPH203 not only in cancer proliferation, but also in tumor stromal activation. These findings will be useful when considering its application in other stroma-abundant carcinomas and its use in combination with other drugs. ABSTRACT: A novel large neutral amino acid transporter 1 (LAT1)-specific inhibitor, JPH203, is expected to cause cancer-specific starvation and possess anti-tumor effects; however, its anti-tumor mechanism for colorectal cancer (CRC) remains unclear. We analyzed LAT family gene expressions in public databases using UCSC Xena and evaluated LAT1 protein expression using immunohistochemistry in 154 cases of surgically resected CRC. We also evaluated mRNA expression using polymerase chain reaction in 10 CRC cell lines. Furthermore, JPH203 treatment experiments were conducted in vitro and in vivo using an allogeneic immune-responsive mouse model with abundant stroma created via the orthotopic transplantation of the mouse-derived CRC cell line CT26 and mesenchymal stem cells. The treatment experiments were followed by comprehensive gene expression analyses with RNA sequencing. Database analyses and immunohistochemistry research on clinical specimens revealed that LAT1 expression was cancer-dominant, and its increase was accompanied by tumor progression. In vitro, JPH203 was effective in an LAT1 expression-dependent manner. In vivo, JPH203 treatment considerably reduced tumor size and metastasis, and RNA sequencing-based pathway analysis showed that not only tumor growth and amino acid metabolism pathways, but also stromal activation-related pathways were suppressed. The results of the RNA sequencing were validated in the clinical specimens, as well as both in vitro and in vivo. LAT1 expression in CRC plays an important role in tumor progression. JPH203 may inhibit the progression of CRC and tumor stromal activity.