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Breast Cancer Research
Published in: Breast Cancer Research 1/2022

01-12-2022 | Lactation | Research article

Multiple roles for Bcl-3 in mammary gland branching, stromal collagen invasion, involution and tumor pathology

Authors: David Carr, Aiman Zein, Josée Coulombe, Tianqi Jiang, Miguel A. Cabrita, Gwendoline Ward, Manijeh Daneshmand, Andrea Sau, M. A. Christine Pratt

Published in: Breast Cancer Research | Issue 1/2022

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Abstract

Background

The Bcl-3 protein is an atypical member of the inhibitor of -κB family that has dual roles as a transcriptional repressor and a coactivator for dimers of NF-κB p50 and p52. Bcl-3 is expressed in mammary adenocarcinomas and can promote tumorigenesis and survival signaling and has a key role in tumor metastasis. In this study, we have investigated the role of Bcl-3 in the normal mammary gland and impact on tumor pathology.

Methods

We utilized bcl-3−/− mice to study mammary gland structure in virgins and during gestation, lactation and early involution. Expression of involution-associated genes and proteins and putative Bcl-3 target genes was examined by qRT-PCR and immunoblot analysis. Cell autonomous branching morphogenesis and collagen I invasion properties of bcl-3−/− organoids were tested in 3D hydrogel cultures. The role of Bcl-3 in tumorigenesis and tumor pathology was also assessed using a stochastic carcinogen-induced mammary tumor model.

Results

Bcl-3−/− mammary glands demonstrated reduced branching complexity in virgin and pregnant mice. This defect was recapitulated in vitro where significant defects in bud formation were observed in bcl-3−/− mammary organoid cultures. Bcl-3−/− organoids showed a striking defect in protrusive collective fibrillary collagen I invasion associated with reduced expression of Fzd1 and Twist2. Virgin and pregnant bcl-3−/− glands showed increased apoptosis and rapid increases in lysosomal cell death and apoptosis after forced weaning compared to WT mice. Bcl-2 and Id3 are strongly induced in WT but not bcl-3−/− glands in early involution. Tumors in WT mice were predominately adenocarcinomas with NF-κB activation, while bcl-3−/− lesions were largely squamous lacking NF-κB and with low Bcl-2 expression.

Conclusions

Collectively, our results demonstrate that Bcl-3 has a key function in mammary gland branching morphogenesis, in part by regulation of genes involved in extracellular matrix invasion. Markedly reduced levels of pro-survival proteins expression in bcl-3 null compared to WT glands 24 h post-weaning indicate that Bcl-3 has a role in moderating the rate of early phase involution. Lastly, a reduced incidence of bcl-3−/− mammary adenocarcinomas versus squamous lesions indicates that Bcl-3 supports the progression of epithelial but not metaplastic cancers.
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Metadata
Title
Multiple roles for Bcl-3 in mammary gland branching, stromal collagen invasion, involution and tumor pathology
Authors
David Carr
Aiman Zein
Josée Coulombe
Tianqi Jiang
Miguel A. Cabrita
Gwendoline Ward
Manijeh Daneshmand
Andrea Sau
M. A. Christine Pratt
Publication date
01-12-2022
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 1/2022
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/s13058-022-01536-w

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