A High-Content Screen Identifies Drugs That Restrict Tumor Cell Extravasation across the Endothelial Barrier

Author(s):Show less -

Hilfenhaus, Georg ^[1] ; Mompeon, Ana ^[2] ; Freshman, Jonathan ^[1] ; Prajapati, Divya P. ^[1] ; Hernandez, Gloria ^[3] ; Freitas, Vanessa M. ^[4] ; Ma, Feiyang ^[3] ; Langenbacher, Adam D. ^[1] ; Mirkov, Snezana ^[2] ; Song, Dana ^[1] ; Cho, Byoung-Kyu ^[5] ; Goo, Young Ah ^[5] ; Pellegrini, Matteo ^{[1, 3]} ; Chen, Jau-Nian ^{[1, 3]} ; Damoiseaux, Robert ^{[6, 7, 8]} ; Iruela-Arispe, M. Luisa ^{[6, 1, 3, 2]}

Affiliation

^[1] Univ Calif Los Angeles, Dept Mol Cell & Dev Biol, Los Angeles, CA - USA

^[2] Northwestern Univ, Feinberg Sch Med, Dept Cell & Dev Biol, Chicago, IL 60611 - USA

^[3] Univ Calif Los Angeles, Mol Biol Inst, Los Angeles, CA - USA

^[4] Univ Sao Paulo, Inst Biomed Sci, Dept Cell & Dev Biol, Sao Paulo - Brazil

^[5] Northwestern Univ, Prote Ctr Excellence, Feinberg Sch Med, Chicago, IL 60611 - USA

^[6] Univ Calif Los Angeles, Jonsson Comprehens Canc Ctr, Los Angeles, CA 90024 - USA

^[7] Univ Calif Los Angeles, Dept Mol & Med Pharmacol, Los Angeles, CA - USA

^[8] Univ Calif Los Angeles, Calif NanoSyst Inst, Los Angeles, CA - USA

• Document type: Journal article
• Source: Cancer Research; v. 81, n. 3, p. 619-633, FEB 1 2021.
• Web of Science Citations: 0

Abstract

Metastases largely rely on hematogenous dissemination of tumor cells via the vascular system and significantly limit prognosis of patients with solid tumors. To colonize distant sites, circulating tumor cells must destabilize the endothelial barrier and transmigrate across the vessel wall. Here we performed a high-content screen to identify drugs that block tumor cell extravasation by testing 3,520 compounds on a transendothelial invasion coculture assay. Hits were further characterized and validated using a series of in vitro assays, a zebrafish model enabling three-dimensional (3D) visualization of tumor cell extravasation, and mouse models of lung metastasis. The initial screen advanced 38 compounds as potential hits, of which, four compounds enhanced endothelial barrier stability while concurrently suppressing tumor cell motility. Two compounds niclosamide and forskolin significantly reduced tumor cell extravasation in zebrafish, and niclosamide drastically impaired metastasis in mice. Because niclosamide had not previously been linked with effects on barrier function, single-cell RNA sequencing uncovered mechanistic effects of the drug on both tumor and endothelial cells. Importantly, niclosamide affected homotypic and heterotypic signaling critical to intercellular junctions, cell-matrix interactions, and cytoskeletal regulation. Proteomic analysis indicated that niclosamide-treated mice also showed reduced levels of kininogen, the precursor to the permeability mediator bradykinin. Our findings designate niclosamide as an effective drug that restricts tumor cell extravasation through modulation of signaling pathways, chemokines, and tumorendothelial cell interactions. Significance: A high-content screen identified niclosamide as an effective drug that restricts tumor cell extravasation by enhancing endothelial barrier stability through modulation of molecular signaling, chemokines, and tumor-endothelial cell interactions. (AU)

• FAPESP's process: 16/19968-3 - Cellular stress on angiogenesis modulation: Biological significance of extracellular vesicles
• Grantee: Vanessa Morais Freitas
• Support Opportunities: Scholarships abroad - Research