Trabectedin is a marine natural product, approved in Europe for the treatment of soft tissue sarcoma and relapsed ovarian cancer. Clinical and experimental evidence indicates that trabectedin is particularly effective against myxoid liposarcomas where response is associated to regression of capillary networks. Here, we investigated the mechanism of the antiangiogenic activity of trabectedin in myxoid liposarcomas. Trabectedin directly targeted endothelial cells, impairing functions relying on extracellular matrix remodeling (invasion and branching morphogenesis) through the upregulation of the inhibitors of matrix metalloproteinases TIMP-1 and TIMP-2. Increased TIMPs synthesis by the tumor microenvironment following trabectedin treatment was confirmed in xenograft models of myxoid liposarcoma. In addition, trabectedin upregulated tumor cell expression of the endogenous inhibitor thrombospondin-1 (TSP-1, a key regulator of angiogenesis-dependent dormancy in sarcoma), in in vivo models of myxoid liposarcomas, in vitro cell lines and primary cell cultures from patients' myxoid liposarcomas. Chromatin Immunoprecipitation analysis showed that trabectedin displaced the master regulator of adipogenesis C/EBPb from the TSP-1 promoter, indicating an association between the upregulation of TSP-1 and induction of adipocytic differentiation program by trabectedin. We conclude that trabectedin inhibits angiogenesis through multiple mechanisms, including directly affecting endothelial cells in the tumor microenvironment-with a potentially widespread activity-and targeting tumor cells' angiogenic activity, linked to a tumor-specific molecular alteration.Trabectedin (Yondelis; ET-743) is a tetrahydroisoquinoline alkaloid isolated from the marine tunicate Ecteinascidia turbinate and now obtained by chemical synthesis.1 It has been approved in Europe for the treatment of advanced or metastatic soft tissue sarcoma and relapsed ovarian cancer, and is currently undergoing phase II trials for several other tumors.Although not yet fully elucidated, its mechanism of action appears different from other antitumor DNA-damaging agents. It binds to N2 of guanine in the minor groove of DNA and interacts with DNA-binding proteins such as DNA repair proteins and transcription factors.1 It affects transcription regulation in a gene-and promoter-dependent fashion in both cancer and normal cells (i.e., macrophages).1-4 As a result trabectedin has a dual antineoplastic effect. It directly targets tumor cells and impairs the recruitment, viability and activity of stroma cells, particularly monocytes/macrophages, 4,5 profoundly affecting the tumor microenvironment, and depriving the tumor of the inflammatory-mediated support.Although trabectedin has shown activity against several types of soft tissue sarcomas, myxoid liposarcomas are the most sensitive to the drug. 6 Myxoid liposarcomas are characterized by chromosomal translocations, most frequently the translocation t(12;16)(q13;p11) leading to the formation of a fusion oncoprotein between FUS (fu...