Development of brain metastases (BM) is associated with neurological decline, morbidity and virtually 100% mortality. BM develop in 10-16% breast cancer patients, with highest incidence for HER2-positive and triple-negative disease. Median survival is 5-22 months depending on treatment and prognostic indicators. There are no standard targeted drug therapies indicated for treatment of BM.
The HER3 (ERBB3) tyrosine kinase is induced in BM compared to matching primary breast and lung cancers, suggesting it could be activated to exploit the abundance of neuregulin (NRG) ligand in the brain. In support of this, we detected very low expression of NRG in clinical samples by RNAseq, and could suppress growth of intracranial MDA-MB-231 breast cancer xenografts by co-grafting a neutralizing Nrg-1 antibody, or treating the mice with Herceptin (i.p.). ERBB3 RNA levels correlate strongly with ERBB2 in clinical samples, but at the protein level HER3 is ubiquitously activated (phosphorylated) in cases comprising a range of HER2 activation levels. IHC analysis of a large archival BM cohort (n=170; 7 primary cancer types) showed strong, complete pHER3 membrane staining in 57.7% cases. Collectively these data suggest that targeting HER3 could be a good therapeutic strategy.
In vitro, we found that Nrg-1 induced phosphorylation cascades involving HER2, HER3, Akt and ERK 1/2 (but not HER4), and increased proliferative and migratory behavior in HER2/3+ breast cancer cell lines. We also tested whether HER3 signaling could promote extracellular protease activity, since protease-mediated modification of the tumour microenvironment is thought to be involved in permeabilising the blood-brain-barrier. Nrg-1 treatment increased expression and activities of Cathepsin B, MMP-2 and MMP-9 in HER2/3+ breast cancer cell lines, and permeabilised a tight human brain microvascular endothelial cell layer in vitro. Treatment with GM6001 (broad spectrum MMP inhibitor), Herceptin or the humanized HER3 monoclonal antibody EV20 opposed Nrg-1-mediated permeability in this assay.