Schöpe Paul Curtis, Heisterkamp Nina, Schütz Devin, Mastrobuoni Guido, Putzker Kerstin, Uhrig Ulrike, Walther Wolfgang, Kempa Stefan, Nazaré Marc, Kobelt Dennis, Stein Ulrike
Experimental and Clinical Research Center, Charité - Universitätsmedizin Berlin, and Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Str. 10, 13125 Berlin, Germany.
Proteomic and Metabolomics Platform, Berlin Institute for Medical Systems Biology (BIMSB), Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany.
Int J Biol Sci. 2025 Jul 11;21(10):4683-4700. doi: 10.7150/ijbs.113805. eCollection 2025.
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 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 (EC < 1 µM; 48 h) and protein level and concomitant restriction of metastatic abilities in two CRC cell lines with a tolerable viability reduction. , 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.
结直肠癌（CRC）转移在全球癌症相关死亡中仍占很大比例。钙结合蛋白S100A4是已知的CRC转移执行者。过去S100A4已与转移形成相关，因此药物干预可降低转移表型。在此，使用转染到HCT116细胞中的S100A4启动子驱动的荧光素酶构建体，对来自EMBL筛选文库的105,600种化合物进行了高通量筛选（HTS），确定了用于S100A4转录抑制的新型化合物。对鉴定出的最有前景的抑制剂进行了S100A4转录抑制、它们对癌细胞伤口愈合、迁移、增殖和活力的影响的测试。随后，在异种移植小鼠模型（HCT116/CMVp-Luc）中测试了领先候选物E12。经过几轮测试，基于2-(4-氟苯磺酰胺基)苯甲酰胺的化合物E12在mRNA（EC<1µM；48小时）和蛋白质水平上对S100A4表达的抑制作用最强，并且在两种CRC细胞系中伴随着转移能力的限制，同时活力降低可耐受。结果表明，转移形成减少，表现为治疗小鼠肝脏中肿瘤的整体生物发光和人类卫星DNA减少。这项研究展示了E12在S100A4靶向转移抑制治疗方面的潜在前景，以改善转移性CRC患者的预后。
