Systems biology analysis reveals NFAT5 as a novel biomarker and master regulator of inflammatory breast cancer.
BACKGROUND
Inflammatory breast cancer (IBC) is the most rare and aggressive variant of breast cancer (BC); however, only a limited number of specific gene signatures with low generalization abilities are available and few reliable biomarkers are helpful to improve IBC classification into a molecularly distinct phenotype. We applied a network-based strategy to gain insight into master regulators (MRs) linked to IBC pathogenesis.


METHODS
In-silico modeling and Algorithm for the Reconstruction of Accurate Cellular Networks (ARACNe) on IBC/non-IBC (nIBC) gene expression data (n = 197) was employed to identify novel master regulators connected to the IBC phenotype. Pathway enrichment analysis was used to characterize predicted targets of candidate genes. The expression pattern of the most significant MRs was then evaluated by immunohistochemistry (IHC) in two independent cohorts of IBCs (n = 39) and nIBCs (n = 82) and normal breast tissues (n = 15) spotted on tissue microarrays. The staining pattern of non-neoplastic mammary epithelial cells was used as a normal control.


RESULTS
Using in-silico modeling of network-based strategy, we identified three top enriched MRs (NFAT5, CTNNB1 or β-catenin, and MGA) strongly linked to the IBC phenotype. By IHC assays, we found that IBC patients displayed a higher number of NFAT5-positive cases than nIBC (69.2% vs. 19.5%; p-value = 2.79 10(-7)). Accordingly, the majority of NFAT5-positive IBC samples revealed an aberrant nuclear expression in comparison with nIBC samples (70% vs. 12.5%; p-value = 0.000797). NFAT5 nuclear accumulation occurs regardless of WNT/β-catenin activated signaling in a substantial portion of IBCs, suggesting that NFAT5 pathway activation may have a relevant role in IBC pathogenesis. Accordingly, cytoplasmic NFAT5 and membranous β-catenin expression were preferentially linked to nIBC, accounting for the better prognosis of this phenotype.


CONCLUSIONS
We provide evidence that NFAT-signaling pathway activation could help to identify aggressive forms of BC and potentially be a guide to assignment of phenotype-specific therapeutic agents. The NFAT5 transcription factor might be developed into routine clinical practice as a putative biomarker of IBC phenotype.
Department of Pathology, Mater Salutis Hospital, Legnago, Italy. remino76@yahoo.it.
Andrea
A
Remo
Department of Science and Technology, University of Sannio, Benevento, Italy. inesimeone@gmail.com.|Qatar Computing Research Institute (QCRI), Qatar Foundation, Doha, Qatar. inesimeone@gmail.com.
Ines
I
Simeone
Department of Science and Technology, University of Sannio, Benevento, Italy. massimo.pancione@unisannio.it.
Massimo
M
Pancione
Department of Pathology and Diagnosis, University of Verona, Verona, Italy. parcesepe.pietro@gmail.com.
Pietro
P
Parcesepe
Department of Molecular Oncology, Institut Paoli-Calmettes, U1068 Inserm, Marseille, France. finettip@ipc.unicancer.fr.
Pascal
P
Finetti
Department of Science and Technology, University of Sannio, Benevento, Italy. luigi.cerulo@gmail.com.|Bioinformatics Laboratory, BIOGEM, Ariano Irpino, Avellino, Italy. luigi.cerulo@gmail.com.
Luigi
L
Cerulo
Qatar Computing Research Institute (QCRI), Qatar Foundation, Doha, Qatar. hbensmail@qf.org.qa.
Halima
H
Bensmail
Department of Molecular Oncology, Institut Paoli-Calmettes, U1068 Inserm, Marseille, France. daniel.birnbaum@inserm.fr.
Daniel
D
Birnbaum
Department Medical Oncology, University of Antwerp, Antwerpen, Belgium. Steven.VanLaere@gza.be.
Steven J
SJ
Van Laere
Department of Science and Technology, University of Sannio, Benevento, Italy. colantuoni@unisannio.it.
Vittorio
V
Colantuoni
Department of Pathology and Diagnosis, University of Verona, Verona, Italy. franco.bonetti@univr.it.
Franco
F
Bonetti
Department of Molecular Oncology, Institut Paoli-Calmettes, U1068 Inserm, Marseille, France. bertuccif@ipc.unicancer.fr.
François
F
Bertucci
Department of Pathology and Diagnosis, University of Verona, Verona, Italy. erminia.manfrin@univr.it.
Erminia
E
Manfrin
Department of Science and Technology, University of Sannio, Benevento, Italy. m.ceccarelli@gmail.com.|Qatar Computing Research Institute (QCRI), Qatar Foundation, Doha, Qatar. m.ceccarelli@gmail.com.
Michele
M
Ceccarelli
D000465:Algorithms; D014408:Biomarkers, Tumor; D002453:Cell Cycle; D015331:Cohort Studies; D019295:Computational Biology; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D006801:Humans; D007150:Immunohistochemistry; D058922:Inflammatory Breast Neoplasms / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D049490:Systems Biology / Q000379:methods*; D014157:Transcription Factors / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism
D014408:Biomarkers, Tumor; C495270:CTNNB1 protein, human; C118125:NFAT5 protein, human; D014157:Transcription Factors; D051176:beta Catenin