Mercier, Anne ElisabethJoubert, Anna MargarethaViallet, JeanDesroches-Castan, AgnesDe Koning, LeanneMabeta, Peaceful LucyHelena, Jolene MichellePepper, Michael SeanLafanechere, Laurence2024-11-132024-11-132024-10Mercier, A.E.; Joubert, A.M.; Prudent, R.; Viallet, J.; DesrochesCastan, A.; De Koning, L.; Mabeta, P.; Helena, J.; Pepper, M.S.; Lafanechère, L. Sulfamoylated Estradiol Analogs Targeting the Actin and Microtubule Cytoskeletons Demonstrate Anti-Cancer Properties In Vitro and In Ovo. Cancers 2024, 16, 2941. https://doi.org/10.3390/cancers16172941.2072-6694 (online)10.3390/cancers16172941http://hdl.handle.net/2263/99034DATA AVAILABITY STATEMENT: All data generated or analysed during this study are included in this published article [and its Supplementary Information File].This article belongs to the Special Issue titled 'Cell Signaling in Cancer and Cancer Therapy'.The microtubule-disrupting agent 2-methoxyestradiol (2-ME) displays anti-tumor and anti-angiogenic properties, but its clinical development is halted due to poor pharmacokinetics. We therefore designed two 2-ME analogs in silico—an ESE-15-one and an ESE-16 one—with improved pharmacological properties. We investigated the effects of these compounds on the cytoskeleton in vitro, and their anti-angiogenic and anti-metastatic properties in ovo. Time-lapse fluorescent microscopy revealed that sub-lethal doses of the compounds disrupted microtubule dynamics. Phalloidin fluorescent staining of treated cervical (HeLa), metastatic breast (MDA-MB-231) cancer, and human umbilical vein endothelial cells (HUVECs) displayed thickened, stabilized actin stress fibers after 2 h, which rearranged into a peripheral radial pattern by 24 h. Cofilin phosphorylation and phosphorylated ezrin/radixin/moesin complexes appeared to regulate this actin response. These signaling pathways overlap with anti-angiogenic, extra-cellular communication and adhesion pathways. Sub-lethal concentrations of the compounds retarded both cellular migration and invasion. Anti-angiogenic and extra-cellular matrix signaling was evident with TIMP2 and P-VEGF receptor-2 upregulation. ESE-15-one and ESE-16 exhibited anti-tumor and anti-metastatic properties in vivo, using the chick chorioallantoic membrane assay. In conclusion, the sulfamoylated 2-ME analogs displayed promising anti-tumor, anti-metastatic, and anti-angiogenic properties. Future studies will assess the compounds for myeloproliferative effects, as seen in clinical applications of other drugs in this class.en© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).MicrotubulesActinAnti-angiogenicMetastasisCofilinEzrin/radixin/moesinChick chorioallantoic membrane assay (CAM)MigrationInvasionCancer2-methoxyestradiol analogsSDG-03: Good health and well-beingArticle