In situ detection of HER2:HER2 and HER2:HER3 protein–protein interactions demonstrates prognostic significance in early breast cancer

Spears, Melanie; Taylor, Karen; Munro, Alison F.; Cunningham, C; Mallon, Elizabeth; Twelves, Chris; Cameron, David; Thomas, Jeremy; Bartlett, John M.S.

doi:10.1007/s10549-011-1606-z

Cited by 63 publications

(60 citation statements)

References 25 publications

(43 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Specifically, HER3 promotes invasion and metastasis through its ability to activate the PI3K pathway [9]. The clinical impact of HER3 is indicated by the observation that increased HER3 expression [10] and the detection HER2/HER3 dimers [11] have prognostic significance in breast cancer.…”

Section: Discussionmentioning

confidence: 99%

Perhexiline promotes HER3 ablation through receptor internalization and inhibits tumor growth

et al. 2015

View full text Add to dashboard Cite

IntroductionHuman epidermal growth factor receptor HER3 has been implicated in promoting the aggressiveness and metastatic potential of breast cancer. Upregulation of HER3 has been found to be a major mechanism underlying drug resistance to EGFR and HER2 tyrosine kinase inhibitors and to endocrine therapy in the treatment of breast cancer. Thus, agents that reduce HER3 expression at the plasma membrane may synergize with current therapies and offer a novel therapeutic strategy to improve treatment.MethodsWe devised an image-based screening platform using membrane localized HER3-YFP to identify small molecules that promote HER3 internalization and degradation. In vitro and in vivo tumor models were used to characterize the signaling effects of perhexiline, an anti-anginal drug, identified by the screening platform.ResultsWe found perhexiline, an anti-anginal drug, selectively internalized HER3, decreased HER3 expression, and subsequently inhibited signaling downstream of HER3. Consistent with these results, perhexiline inhibited breast cancer cell proliferation in vitro and tumor growth in vivo.ConclusionsThis is the first demonstration that HER3 can be targeted with small molecules by eliminating it from the cell membrane. The novel approach used here led to the discovery that perhexiline ablates HER3 expression, and offers an opportunity to identify HER3 ablation modulators as innovative therapeutics to improve survival in breast cancer patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-015-0528-9) contains supplementary material, which is available to authorized users.

show abstract

Section: Discussionmentioning

confidence: 99%

Perhexiline promotes HER3 ablation through receptor internalization and inhibits tumor growth

et al. 2015

View full text Add to dashboard Cite

show abstract

“…As HER3 has been shown to be the preferred interaction partner of HER2 [21, 22] and as pertuzumab is targeted against this interaction, several groups have attempted to quantify HER2-HER3 dimerisation in different cell types and xenograft models [23] as well as in FFPE samples [23, 24]. These studies use either the Proximity Ligation Assay (PLA) or the VeraTag assay, which can both detect proteins in close proximity, inferring interaction.…”

Section: Introductionmentioning

confidence: 99%

“…The equivalent distance can be up to 300 nm for the VeraTag assay [26, 27]. Using the PLA technique (primary plus secondary antibodies conjugated to oligonucleotides), high levels of HER2-HER2 and HER2-HER3 protein proximity have previously been shown to be correlated with HER2 amplification/overexpression, using 88 and 74 cases of human breast carcinomas for HER2-HER2 and HER2-HER3 PLAs, respectively [24]. …”

Section: Introductionmentioning

confidence: 99%

HER2-HER3 dimer quantification by FLIM-FRET predicts breast cancer metastatic relapse independently of HER2 IHC status

et al. 2016

View full text Add to dashboard Cite

Overexpression of HER2 is an important prognostic marker, and the only predictive biomarker of response to HER2-targeted therapies in invasive breast cancer. HER2-HER3 dimer has been shown to drive proliferation and tumor progression, and targeting of this dimer with pertuzumab alongside chemotherapy and trastuzumab, has shown significant clinical utility. The purpose of this study was to accurately quantify HER2-HER3 dimerisation in formalin fixed paraffin embedded (FFPE) breast cancer tissue as a novel prognostic biomarker.FFPE tissues were obtained from patients included in the METABRIC (Molecular Taxonomy of Breast Cancer International Consortium) study. HER2-HER3 dimerisation was quantified using an improved fluorescence lifetime imaging microscopy (FLIM) histology-based analysis. Analysis of 131 tissue microarray cores demonstrated that the extent of HER2-HER3 dimer formation as measured by Förster Resonance Energy Transfer (FRET) determined through FLIM predicts the likelihood of metastatic relapse up to 10 years after surgery (hazard ratio 3.91 (1.61–9.5), p = 0.003) independently of HER2 expression, in a multivariate model. Interestingly there was no correlation between the level of HER2 protein expressed and HER2-HER3 heterodimer formation. We used a mathematical model that takes into account the complex interactions in a network of all four HER proteins to explain this counterintuitive finding.Future utility of this technique may highlight a group of patients who do not overexpress HER2 protein but are nevertheless dependent on the HER2-HER3 heterodimer as driver of proliferation. This assay could, if validated in a group of patients treated with, for instance pertuzumab, be used as a predictive biomarker to predict for response to such targeted therapies.

show abstract

“…This overexpression is thought to cause activation of c-erbB2 by ligand-independent homodimerisation (contrasting with its normal function in ligand-induced heterodimerisation with other EGF receptor family members) (13). In our model, high c-erbB2 signalling is induced in an immortalised human luminal mammary epithelial cell line [HB2, originally developed in the lab of Dr Joyce Taylor-Papadimitriou (14)] by means of a hybrid RTK, ‘trk-neu’.…”

Section: Introductionmentioning

confidence: 99%

Loss of E-cadherin expression is not a prerequisite for c-erbB2-induced epithelial-mesenchymal transition

Nilsson¹,

Akhtar²,

Kannius-Janson³

et al. 2014

International Journal of Oncology

View full text Add to dashboard Cite

Recent research into the mechanisms of tumour cell invasiveness has highlighted the parallels between carcinogenesis and epithelial-mesenchymal transition (EMT), originally described as a developmental transdifferentiation program but also implicated in fibrosis and cancer. In a model system for mammary carcinogenesis, we previously observed that induced signalling from a homodimer of the c-erbB2 (HER2) receptor tyrosine kinase in an initially non-malignant mammary cell line caused EMT where i) cell scattering occurred before downregulation of the cell-cell adhesion molecule E-cadherin and ii) the progress of EMT was dramatically delayed when cells were grown at high density. Here, we have further analysed these phenomena. Ectopic expression of E-cadherin concomitant with c-erbB2 signalling was unable to impede the progression of EMT, suggesting that E-cadherin downregulation is not required for EMT. Furthermore, fibroblast-like cells isolated after EMT induced in the presence or absence of ectopic E-cadherin expression showed highly similar morphology and vimentin expression. E-cadherin expressed in these fibroblastic cells had a subcellular localisation similar to that found in epithelial cells, but it exhibited a much weaker attachment to the cytoskeleton, suggesting cytoskeletal rearrangements as an important mechanism in EMT-associated cell scattering. We also investigated whether density-dependent inhibition of EMT is mediated by E-cadherin as a sensor for cell-cell contact, by expressing dominant-negative E-cadherin. While expression of this mutant weakened cell-cell adhesion, it failed to facilitate EMT at high cell densities. These results indicate that loss of E-cadherin expression is a consequence rather than a cause of c-erbB2-induced EMT and that density-dependent inhibition of EMT is not mediated by E-cadherin signalling.

show abstract

In situ detection of HER2:HER2 and HER2:HER3 protein–protein interactions demonstrates prognostic significance in early breast cancer

Cited by 63 publications

References 25 publications

Perhexiline promotes HER3 ablation through receptor internalization and inhibits tumor growth

Perhexiline promotes HER3 ablation through receptor internalization and inhibits tumor growth

HER2-HER3 dimer quantification by FLIM-FRET predicts breast cancer metastatic relapse independently of HER2 IHC status

Loss of E-cadherin expression is not a prerequisite for c-erbB2-induced epithelial-mesenchymal transition

Contact Info

Product

Resources

About