# Novel High-Throughput Screen Identified S100A4 Inhibitors for Anti-Metastatic Therapy

## TL;DR

A new compound, E12, was found to inhibit S100A4, a protein linked to colorectal cancer metastasis, potentially improving treatment for advanced cancer patients.

## Contribution

A high-throughput screen identified E12 as a novel S100A4 inhibitor with anti-metastatic effects in colorectal cancer.

## Key findings

- E12 inhibited S100A4 expression at mRNA and protein levels in CRC cell lines.
- E12 reduced metastatic abilities and tumor bioluminescence in a xenograft mouse model.
- E12 showed tolerable viability reduction in cancer cells with anti-metastatic potential.

## Abstract

Colorectal cancer (CRC) metastasis continues to account for a substantial proportion of cancer-related deaths worldwide. Calcium-binding protein S100A4 is a known executor of CRC metastasis. S100A4 has been correlated to metastasis formation in the past, and therefore pharmaceutical intervention reduces the metastatic phenotype. Herein, a high-throughput screen (HTS) of 105,600 compounds from the EMBL screening library using an S100A4 promoter-driven luciferase construct transfected into HCT116 cells identified novel compounds for S100A4 transcriptional inhibition. The most promising inhibitors identified were tested for S100A4 transcriptional inhibition, their impact on wound healing, migration, proliferation and viability of cancer cells. Subsequently, the leading candidate E12 was tested in vivo in a xenograft mouse model (HCT116/CMVp- Luc). After several testing rounds, E12 a 2-(4-fluorobenzenesulfonamido)benzamide-based compound showed the strongest inhibition of S100A4 expression at mRNA (EC50 < 1 µM; 48 h) and protein level and concomitant restriction of metastatic abilities in two CRC cell lines with a tolerable viability reduction. In vivo, a reduction in metastasis formation was demonstrated, displayed by reduced overall bioluminescence of tumors and human satellite DNA in the liver of treated mice. This study exhibited E12's promising potential for S100A4 targeted metastasis inhibition therapy to improve the outcome of metastasized CRC patients.