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      Dynamic architectural interplay between leucocytes and mammary epithelial cells

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          Abstract

          The adult mammary gland undergoes dynamic changes during puberty and the postnatal developmental cycle. The mammary epithelium is composed of a bilayer of outer basal, or myoepithelial, cells and inner luminal cells, the latter lineage giving rise to the milk‐producing alveolar cells during pregnancy. These luminal alveolar cells undergo Stat3‐mediated programmed cell death following the cessation of lactation. It is established that immune cells in the microenvironment of the gland have a role to play both in the ductal outgrowth during puberty and in the removal of dead cells and remodelling of the stroma during the process of postlactational regression. However, most studies have focussed on the role of the stromal immune cell compartment or have quantified immune cell populations in tissue extracts. Our recent development of protocols for deep imaging of the mammary gland in three dimensions (3D) has enabled the architectural relationship between immune cells and the epithelium to be examined in detail, and we have discovered a surprisingly dynamic relationship between the basal epithelium and leucocytes. Furthermore, we have observed morphological changes in the myoepithelial cells, as involution progresses, which were not revealed by previous work in 2D tissue sections and whole tissue. This dynamic architecture suggests a role for myoepithelial cells in the orderly progression of involution. We conclude that deep imaging of mammary gland and other tissues is essential for analysing complex interactions between cellular compartments.

          Abstract

          In this study by Christine Watson and colleagues, deep 3D‐imaging was used to investigate changes in the architecture of myoepithelial cells and leukocytes in mammary ducts and lobuloalveolar structures. Leukocytes intercalate between ductal myoepithelial and luminal cells and surprisingly, closely associate with myoepithelial cells during lactation and involution, adopting their shape. Myoepithelial cells contract progressively as involution proceeds and shrink back towards the ducts, possibly providing a barrier to the dissemination of milk and dying luminal cells.

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          Mammary development in the embryo and adult: a journey of morphogenesis and commitment.

          Mammary gland development occurs through distinctive stages throughout embryonic and pubertal development and reproductive life. At each stage, different signals are required to induce changes in both the epithelium and the surrounding mesenchyme/stroma. Recent studies have provided new insights into the origin, specification and fate of mammary stem and progenitor cells and into how the differentiated lineages that comprise the functional mammary gland are determined. The development of new tools and culture techniques has also enabled the factors that influence branching morphogenesis in the embryonic and pubertal gland to be identified. A surprising recent discovery has been that mammary epithelial cells commit to differentiated lineages using the same signalling pathways that regulate lineage determination in T helper cells.
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            Suppression of epithelial apoptosis and delayed mammary gland involution in mice with a conditional knockout of Stat3.

            Mammary gland involution is characterized by extensive apoptosis of the epithelial cells. At the onset of involution, Stat3 is specifically activated. To address the function of this signaling molecule in mammary epithelial apoptosis, we have generated a conditional knockout of Stat3 using the Cre-lox recombination system. Following weaning, a decrease in apoptosis and a dramatic delay of involution occurred in Stat3 null mammary tissue. Involution is normally associated with a significant increase in IGFBP-5 levels. This was observed in control glands, but not in the absence of Stat3. IGFBP-5 has been suggested to induce apoptosis by sequestering IGF-1 to casein micelles, thereby inhibiting its survival function. Our findings suggest that IGFBP-5 is a direct or indirect target for Stat3 and its upregulation is essential to normal involution. No marked differences were seen in the regulation of Stat5, Bcl-x(L), or Bax in the absence of Stat3. Precocious activation of Stat1 and increases in levels of p53 and p21 occurred and may act as compensatory mechanisms for the eventual initiation of involution observed in Stat3 null mammary glands. This is the first demonstration of the importance of a Stat factor in signaling the initiation of physiological apoptosis in vivo.
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              Stat3 controls lysosomal-mediated cell death in vivo.

              It is well established that lysosomes play an active role during the execution of cell death. A range of stimuli can lead to lysosomal membrane permeabilization (LMP), thus inducing programmed cell death without involvement of the classical apoptotic programme. However, these lysosomal pathways of cell death have mostly been described in vitro or under pathological conditions. Here we show that the physiological process of post-lactational regression of the mammary gland is accomplished through a non-classical, lysosomal-mediated pathway of cell death. We found that, during involution, lysosomes in the mammary epithelium undergo widespread LMP. Furthermore, although cell death through LMP is independent of executioner caspases 3, 6 and 7, it requires Stat3, which upregulates the expression of lysosomal proteases cathepsin B and L, while downregulating their endogenous inhibitor Spi2A (ref. 8). Our findings report a previously unknown, Stat3-regulated lysosomal-mediated pathway of cell death under physiological circumstances. We anticipate that these findings will be of major importance in the design of treatments for cancers such as breast, colon and liver, where cathepsins and Stat3 are commonly overexpressed and/or hyperactivated respectively. © 2011 Macmillan Publishers Limited. All rights reserved.
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                Author and article information

                Contributors
                cjw53@cam.ac.uk
                Journal
                FEBS J
                FEBS J
                10.1111/(ISSN)1742-4658
                FEBS
                The Febs Journal
                John Wiley and Sons Inc. (Hoboken )
                1742-464X
                1742-4658
                29 November 2019
                January 2020
                : 287
                : 2 ( doiID: 10.1111/febs.v287.2 )
                : 250-266
                Affiliations
                [ 1 ] Department of Pathology University of Cambridge UK
                [ 2 ] Department of Veterinary Medicine Cambridge UK
                Author notes
                [*] [* ] Correspondence

                C. J. Watson, Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK

                Tel: +44 01223 333725

                E‐mail: cjw53@ 123456cam.ac.uk

                Author information
                https://orcid.org/0000-0002-3331-1249
                https://orcid.org/0000-0002-8548-5902
                Article
                FEBS15126
                10.1111/febs.15126
                7003847
                31691481
                aa62000c-0f4d-431e-bf2d-33d0cb7bdd29
                © 2019 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 13 June 2019
                : 19 August 2019
                : 04 November 2019
                Page count
                Figures: 6, Tables: 0, Pages: 17, Words: 8836
                Funding
                Funded by: Medical Research Council , open-funder-registry 10.13039/501100000265;
                Award ID: MR/N022963/1
                Categories
                Editor's Choice
                Editor's Choice
                Custom metadata
                2.0
                January 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.5 mode:remove_FC converted:06.02.2020

                Molecular biology
                imaging,involution,leucocytes,mammary gland
                Molecular biology
                imaging, involution, leucocytes, mammary gland

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