The effect of U46619 and CPI211 on migration and invasion of 4T1 and MDA-MB-231 tumor cells through Matrigel-coated transwells was assessed. Tumor cells were seeded in the upper chambers of a Matrigel-coated transwell insert in serum-free media. Cell migration towards 1% serum in the lower chamber was assessed by crystal violet staining of the lower side of the transwell filter. Digital images of stained transwell filters were used to count the number of migrated cells. Representative images are shown (A). Each data point shown (B) represents the number of cells migrating to the lower side of the filter. Midlines are the average {plus minus} S.D., N = 4 (4T1) and 5 (MDA-MB-231), each assessed in triplicate.
Funding
NIH
National Center for Advancing Translational Sciences
Congressionally Directed Medical Research Programs
ARTICLE ABSTRACT
Although new drug discoveries are revolutionizing cancer treatments, repurposing existing drugs would accelerate the timeline and lower the cost for bringing treatments to cancer patients. Our goal was to repurpose CPI211, a potent and selective antagonist of the thromboxane A2-prostanoid receptor (TPr), a G-protein–coupled receptor that regulates coagulation, blood pressure, and cardiovascular homeostasis. To identify potential new clinical indications for CPI211, we performed a phenome-wide association study (PheWAS) of the gene encoding TPr, TBXA2R, using robust deidentified health records and matched genomic data from more than 29,000 patients. Specifically, PheWAS was used to identify clinical manifestations correlating with a TBXA2R single-nucleotide polymorphism (rs200445019), which generates a T399A substitution within TPr that enhances TPr signaling. Previous studies have correlated 200445019 with chronic venous hypertension, which was recapitulated by this PheWAS analysis. Unexpectedly, PheWAS uncovered an rs200445019 correlation with cancer metastasis across several cancer types. When tested in several mouse models of metastasis, TPr inhibition using CPI211 potently blocked spontaneous metastasis from primary tumors, without affecting tumor cell proliferation, motility, or tumor growth. Further, metastasis following intravenous tumor cell delivery was blocked in mice treated with CPI211. Interestingly, TPr signaling in vascular endothelial cells induced VE-cadherin internalization, diminished endothelial barrier function, and enhanced transendothelial migration by tumor cells, phenotypes that were decreased by CPI211. These studies provide evidence that TPr signaling promotes cancer metastasis, supporting the study of TPr inhibitors as antimetastatic agents and highlighting the use of PheWAS as an approach to accelerate drug repurposing.
