TY - JOUR
T1 - A High-Content Screen Identifies Drugs That Restrict Tumor Cell Extravasation across the Endothelial Barrier
AU - Hilfenhaus, Georg
AU - Mompeon, Ana
AU - Freshman, Jonathan
AU - Prajapati, Divya P.
AU - Hernandez, Gloria
AU - Freitas, Vanessa M.
AU - Ma, Feiyang
AU - Langenbacher, Adam D.
AU - Mirkov, Snezana
AU - Song, Dana
AU - Cho, Byoung Kyu
AU - Goo, Young Ah
AU - Pellegrini, Matteo
AU - Chen, Jau Nian
AU - Damoiseaux, Robert
AU - Iruela-Arispe, Maria Luisa
N1 - Funding Information:
The authors thank Douglas Hanahan for providing the pancreatic cancer cell lines; Austin Quach for support with R; the UCLA Zebrafish Core Facility; Sequencing Core Facility; Broad Stem Cell Institute Flow Cytometry Core; Integrated Molecular Technologies Core. All high-throughput screening experiments were performed at the UCLA Molecular Screening Shared Resource (supported by Comprehensive Cancer Center Grant NIH/NCI P30CA016042). Proteomics services were performed by the Northwestern Proteomics Core Facility, generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center, instrumentation award (S10OD025194) from NIH Office of Director, and the National Resource for Translational and Developmental Proteomics supported by P41 GM108569. Metabolomics experiments were performed by the Metabolomics Core Facility at Feinberg School of Medicine at Northwestern University. This work was supported by NIH R01CA197943 and R35HL140014 (to M.L. Iruela-Arispe), R01 HL140472 (to J.-N. Chen), and a fellowship of the German Research Foundation (DFG; #HI1727/1?1 to G. Hilfenhaus) and of the Sao Paulo Research Foundation (FAPESP; #2016/19968?3 to V. Freitas).
Funding Information:
The authors thank Douglas Hanahan for providing the pancreatic cancer cell lines; Austin Quach for support with R; the UCLA Zebrafish Core Facility; Sequencing Core Facility; Broad Stem Cell Institute Flow Cytometry Core; Integrated Molecular Technologies Core. All high-throughput screening experiments were performed at the UCLA Molecular Screening Shared Resource (supported by Comprehensive Cancer Center Grant NIH/NCI P30CA016042). Proteomics services were performed by the Northwestern Proteomics Core Facility, generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center, instrumentation award (S10OD025194) from NIH Office of Director, and the National Resource for Translational and Developmental Proteomics supported by P41 GM108569. Metabolomics experiments were performed by the Metabolomics Core Facility at Feinberg School of Medicine at Northwestern University. This work was supported by NIH R01CA197943 and R35HL140014 (to M.L. Iruela-Arispe), R01 HL140472 (to J.-N. Chen), and a fellowship of the German Research Foundation (DFG; #HI1727/1–1 to G. Hilfenhaus) and of the Sao Paulo Research Foundation (FAPESP; #2016/19968–3 to V. Freitas).
Publisher Copyright:
©2020 American Association for Cancer Research.
PY - 2021/2/1
Y1 - 2021/2/1
N2 - 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 tumor–endothelial 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.
AB - 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 tumor–endothelial 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.
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U2 - 10.1158/0008-5472.CAN-19-3911
DO - 10.1158/0008-5472.CAN-19-3911
M3 - Article
C2 - 33218969
AN - SCOPUS:85100376083
VL - 81
SP - 619
EP - 633
JO - Cancer Research
JF - Cancer Research
SN - 0008-5472
IS - 3
ER -