38 results on '"Fre, S."'
Search Results
2. Proffered Paper: Mechanical induction of the tumourogenic beta-catenin pathway by tumour growth pressure
- Author
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Fernández-Sánchez, M.E., primary, Barbier, S., additional, Whitehead, J., additional, Béalle, G., additional, Michel, A., additional, Latorre-Ossa, H., additional, Rey, C., additional, Fouassier, L., additional, Claperon, A., additional, Brullé, L., additional, Girard, E., additional, Servant, N., additional, Rio-Frio, T., additional, Marie, H., additional, Lesieur, S., additional, Housset, C., additional, Gennisson, J.-L., additional, Tanter, M., additional, Ménager, C., additional, Fre, S., additional, Robine, S., additional, and Farge, E., additional
- Published
- 2016
- Full Text
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3. Conditional Mouse Models of Cancer
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Vignjevic, D., primary, Fre, S., additional, Louvard, D., additional, and Robine, S., additional
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4. N=2 SUPERGRAVITY AND N=2 SUPERYANG-MILLS THEORY ON GENERAL SCALAR MANIFOLDS: SYMPLECTIC COVARIANCE, GAUGINGS AND THE MOMENTUM MAP
- Author
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Andrianopoli, Laura Maria, Bertolini, M., Ceresole, Anna Teresa, D'Auria, Riccardo, Fre, S. FERRARA P., and Magri, T.
- Published
- 1997
5. Recognition specificity of individual EH domains of mammals and yeast.
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Paoluzi, S, Castagnoli, L, Lauro, I, Salcini, A E, Coda, L, Fre', S, Confalonieri, S, Pelicci, P G, Di Fiore, P P, Cesareni, G, Paoluzi, S, Castagnoli, L, Lauro, I, Salcini, A E, Coda, L, Fre', S, Confalonieri, S, Pelicci, P G, Di Fiore, P P, and Cesareni, G
- Abstract
Udgivelsesdato: 1998-Nov-16, The Eps homology (EH) domain is a recently described protein binding module that is found, in multiple or single copies, in several proteins in species as diverse as human and yeast. In this work, we have investigated the molecular details of recognition specificity mediated by this domain family by characterizing the peptide-binding preference of 11 different EH domains from mammal and yeast proteins. Ten of the eleven EH domains could bind at least some peptides containing an Asn-Pro-Phe (NPF) motif. By contrast, the first EH domain of End3p preferentially binds peptides containing an His-Thr/Ser-Phe (HT/SF) motif. Domains that have a low affinity for the majority of NPF peptides reveal some affinity for a third class of peptides that contains two consecutive amino acids with aromatic side chains (FW or WW). This is the case for the third EH domain of Eps15 and for the two N-terminal domains of YBL47c. The consensus sequences derived from the peptides selected from phage-displayed peptide libraries allows for grouping of EH domains into families that are characterized by different NPF-context preference. Finally, comparison of the primary sequence of EH domains with similar or divergent specificity identifies a residue at position +3 following a conserved tryptophan, whose chemical characteristics modulate binding preference.
- Published
- 1998
6. Conditional Mouse Models of Cancer.
- Author
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Starke, K., Born, G. V. R., Duckles, S., Eichelbaum, M., Ganten, D., Hofmann, F., Rosenthal, W., Rubanyi, G., Feil, Robert, Metzger, Daniel, Vignjevic, D., Fre, S., Louvard, D., and Robine, S.
- Abstract
The development of inducible and conditional technologies allowed us to generate transgenic mouse models that faithfully recapitulate human tumorigenesis. It is possible to control, in time and space, the development of tumors in almost every mouse tissue. The result is that now we have available mouse models for all major human cancers. Novel noninvasive approaches to tumor imaging will enable us to follow tumor development and metastasis in vivo, as well as the effects of candidate therapeutic drugs. Such new generation tumor models, which accurately emulate the disease state in situ, should provide a useful platform with which to experimentally test drugs targeted to specific gene products, or combinations of genes that control rate-limiting steps of tumor development. In this review, we focus on the different mouse models for colon cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2007
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7. Mechanical induction of the oncogenic Beta-catenin pathway by tumour growth pressure
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Barbier, S., Sanchez, M. Fernandez, Whitehead, J., Bealle, G., Michel, A., Latorre-Ossa, H., Rey, C., Fouassier, L., Claperon, A., Brulle, L., Girard, E., Servant, N., Rio-Frio, T., Marie, H., Lesieur, S., Housset, C., Genisson, J., Mickael Tanter, Menager, C., Fre, S., Robine, S., and Farge, E.
8. Notch signals and intestinal stem cells: from crypt homeostasis to colon cancer.
- Author
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Fre, S.
- Subjects
- *
STEM cells , *CELL proliferation , *NOTCH genes - Abstract
In adult organisms, tissues are maintained and repaired by stem cells, which divide and differentiate to generate more specialized progeny. The mechanisms that control the balance between self-renewal and differentiation promise fundamental insights into the origin and design of multi-cellular organisms. A tissue that is particularly suited to approach these questions is the intestinal epithelium, as it consists of a monolayer of epithelial cells that endlessly divide, migrate while differentiating and are replaced to ensure continuous and fast cell renewal throughout adult life. In this tissue, somatic stem cells represent crucial elements that govern tissue remodeling and homeostasis and the combined work of several laboratories has provided evidence for the existence of at least two different stem cell populations in the small intestinal crypt. In this program, we focus on the Notch signaling pathway as a new promising marker to study gut stem cell physiology. Our observations indicate that the Notch1 receptor is expressed in both crypt stem cell populations, implying that it can mark either one of the two in vivo and providing a precious tool to dissect the hierarchy between these different stem cells. Notch signaling plays an evolutionarily conserved role in metazoans, which basically consists in mediating cell coordination in development and during adult tissue homeostasis, through the profound effects it has on stem cells. The precise identity of the cells in which Notch signaling is active is still unclear, mainly due to the lack of reliable tools to investigate Notch expression in vivo. For this purpose, we have recently developed and characterized a novel roster of unique transgenic mice that permit to assess Notch expression and activation in vivo in an unprecedented fashion. Thanks to these novel mice, we have been able to formally show that the expression of the Notch1 receptor and of its transcriptional target Hes1 identifies crypt stem cells. We are now using these mouse models to dissect the hierarchy between stem cell populations in the small intestine, while expanding our knowledge on colonic stem cells, that remain poorly studied. In addition, we study the dynamic behavior of stem cells during regeneration upon injury. The critical role played by Notch signaling in intestinal renewal and differentiation, as well as in tumorigenesis in this tissue, is exemplified by the vast number of reports on this topic appeared in recent years (i.e. [1-6]). In the past five years, the Notch signaling pathway has emerged as an essential regulator of intestinal homeostasis; indeed Notch signals can control the segregation of each mature lineage from undifferentiated progenitor cells, and they are instrumental for maintaining the proliferating intestinal cell pool. When Notch signaling is inhibited, all crypt cells cease to proliferate and differentiate into secretory cells [4]. Reciprocally, we have shown that gain of Notch function in the developing intestinal epithelium dramatically impairs cell differentiation and increases the proportion of dividing cells [2]. The role of Notch in promoting intestinal proliferation requires Wnt signals, whereas it specifies cell fate independently of Wnt [5]. Importantly, our work has shown that Notch acts in synergy with the Wnt pathway to induce intestinal polyps [5]. The accumulated evidence in both systems supports a role for Notch in expanding a potentially malignant cell population and hence increasing the chances of a tumorigenic event. It is indeed widely accepted that the primary events in tumourigenesis are linked to stem cell transformation: the process of tumor development is thought to initially affect normal stem cells or closely related early progenitors. In order to establish the identity of Notch1-expressing cells within a tumor and to follow their fate in vivo, we have crossed N1-CreERT2/GFP mice to Apc heterozygous animals, which give rise to intestinal tumors by stochastic loss of heterozygosity (LOH) at the Apc locus. Our novel genetic fate-mapping system will consent to identify which cells within a tumor express the Notch1 receptor, as well as to follow the progeny of the Notch-expressing tumor cells. Our preliminary results show that only a small fraction of tumor cells expresses the N1cre-driven GFP reporter. These experiments will allow us to highlight in vivo which and how many cells within a tumor present Notch activity. Our research aims at understanding the molecular features of adult stem cells, as well as the mechanisms by which Notch signaling controls the fate of specific cell populations during regeneration and tumourigenesis. We believe that the identification of common organizational principles of tissue architecture is an essential step in order to develop safe and efficacious applications of stem cells in regenerative medicine. Fre, S., et al., Notch lineages and activity in intestinal stem cells determined by a new set of knock-in mice. [ABSTRACT FROM AUTHOR]
- Published
- 2013
9. 7 - Proffered Paper: Mechanical induction of the tumourogenic beta-catenin pathway by tumour growth pressure.
- Author
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Fernández-Sánchez, M.E., Barbier, S., Whitehead, J., Béalle, G., Michel, A., Latorre-Ossa, H., Rey, C., Fouassier, L., Claperon, A., Brullé, L., Girard, E., Servant, N., Rio-Frio, T., Marie, H., Lesieur, S., Housset, C., Gennisson, J.-L., Tanter, M., Ménager, C., and Fre, S.
- Published
- 2016
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10. Alpha-6 integrin deletion delays the formation of Brca1/p53-deficient basal-like breast tumors by restricting luminal progenitor cell expansion.
- Author
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Faraldo MM, Romagnoli M, Wallon L, Dubus P, Deugnier MA, and Fre S
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- Animals, Female, Mice, Humans, Neoplastic Stem Cells metabolism, Neoplastic Stem Cells pathology, Cell Proliferation, Stem Cells metabolism, Gene Deletion, Cell Transformation, Neoplastic genetics, Cell Transformation, Neoplastic metabolism, Integrin alpha6 metabolism, Integrin alpha6 genetics, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism, BRCA1 Protein genetics, BRCA1 Protein metabolism, Breast Neoplasms pathology, Breast Neoplasms genetics, Breast Neoplasms metabolism
- Abstract
Background: The aberrant amplification of mammary luminal progenitors is at the origin of basal-like breast cancers associated with BRCA1 mutations. Integrins mediate cell-matrix adhesion and transmit mechanical and chemical signals that drive epithelial stem cell functions and regulate tumor progression, metastatic reactivation, and resistance to targeted therapies. Consistently, we have recently shown that laminin-binding integrins are essential for the expansion and differentiation of mammary luminal progenitors in physiological conditions. As over-expression of the laminin-binding α6 integrin (Itgα6) is associated with poor prognosis and reduced survival in breast cancer, we here investigate the role of Itgα6 in mammary tumorigenesis., Methods: We used Blg-Cre; Brca1
F/F ; Trp53F/F mice, a model that phenocopies human basal-like breast cancer with BRCA1 mutations. We generated mutant mice proficient or deficient in Itgα6 expression and followed tumor formation. Mammary tumors and pretumoral tissues were characterized by immunohistochemistry, flow cytometry, RT-qPCR, Western blotting and organoid cultures. Clonogenicity of luminal progenitors from preneoplastic glands was studied in 3D Matrigel cultures., Results: We show that Itga6 deletion favors activation of p16 cell cycle inhibitor in the preneoplastic tissue. Subsequently, the amplification of luminal progenitors, the cell of origin of Brca1-deficient tumors, is restrained in Itgα6-deficient gland. In addition, the partial EMT program operating in Brca1/p53-deficient epithelium is attenuated in the absence of Itgα6. As a consequence of these events, mammary tumor formation is delayed in Itgα6-deficient mice. After tumor formation, the lack of Itgα6 does not affect tumor growth but rather alters their differentiation, resulting in reduced expression of basal cell markers., Conclusions: Our data indicate that Itgα6 has a pro-tumorigenic role in Blg-Cre; Brca1F/F ; Trp53F/F mice developing basal-like mammary tumors. In particular, we reveal that Itgα6 is required for the luminal progenitor expansion and the aberrant partial EMT program that precedes the formation of BRCA1 deficient tumors., (© 2024. The Author(s).)- Published
- 2024
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11. Spatially distinct epithelial and mesenchymal cell subsets along progressive lineage restriction in the branching embryonic mammary gland.
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Carabaña C, Sun W, Veludo Ramos C, Huyghe M, Perkins M, Maillot A, Journot R, Hartani F, Faraldo MM, Lloyd-Lewis B, and Fre S
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- Animals, Mice, Female, Cell Differentiation, Multipotent Stem Cells cytology, Multipotent Stem Cells metabolism, Fibroblast Growth Factor 10 metabolism, Fibroblast Growth Factor 10 genetics, Morphogenesis, Single-Cell Analysis, Mesoderm cytology, Mesoderm metabolism, Mesoderm embryology, Mammary Glands, Animal cytology, Mammary Glands, Animal embryology, Mammary Glands, Animal metabolism, Cell Lineage, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells metabolism, Epithelial Cells cytology, Epithelial Cells metabolism
- Abstract
How cells coordinate morphogenetic cues and fate specification during development remains a fundamental question in organogenesis. The mammary gland arises from multipotent stem cells (MaSCs), which are progressively replaced by unipotent progenitors by birth. However, the lack of specific markers for early fate specification has prevented the delineation of the features and spatial localization of MaSC-derived lineage-committed progenitors. Here, using single-cell RNA sequencing from E13.5 to birth, we produced an atlas of matched mouse mammary epithelium and mesenchyme and reconstructed the differentiation trajectories of MaSCs toward basal and luminal fate. We show that murine MaSCs exhibit lineage commitment just prior to the first sprouting events of mammary branching morphogenesis at E15.5. We identify early molecular markers for committed and multipotent MaSCs and define their spatial distribution within the developing tissue. Furthermore, we show that the mammary embryonic mesenchyme is composed of two spatially restricted cell populations, and that dermal mesenchyme-produced FGF10 is essential for embryonic mammary branching morphogenesis. Altogether, our data elucidate the spatiotemporal signals underlying lineage specification of multipotent MaSCs, and uncover the signals from mesenchymal cells that guide mammary branching morphogenesis., (© 2024. The Author(s).)
- Published
- 2024
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12. Extracting, filtering and simulating cellular barcodes using CellBarcode tools.
- Author
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Sun W, Perkins M, Huyghe M, Faraldo MM, Fre S, Perié L, and Lyne AM
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- Reproducibility of Results, Sequence Analysis, DNA methods, Polymerase Chain Reaction, DNA Barcoding, Taxonomic methods, DNA genetics
- Abstract
Identifying true DNA cellular barcodes among polymerase chain reaction and sequencing errors is challenging. Current tools are restricted in the diversity of barcode types supported or the analysis strategies implemented. As such, there is a need for more versatile and efficient tools for barcode extraction, as well as for tools to investigate which factors impact barcode detection and which filtering strategies to best apply. Here we introduce the package CellBarcode and its barcode simulation kit, CellBarcodeSim, that allows efficient and versatile barcode extraction and filtering for a range of barcode types from bulk or single-cell sequencing data using a variety of filtering strategies. Using the barcode simulation kit and biological data, we explore the technical and biological factors influencing barcode identification and provide a decision tree on how to optimize barcode identification for different barcode settings. We believe that CellBarcode and CellBarcodeSim have the capability to enhance the reproducibility and interpretation of barcode results across studies., (© 2024. The Author(s).)
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- 2024
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13. Fibroblast-induced mammary epithelial branching depends on fibroblast contractility.
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Sumbal J, Fre S, and Sumbalova Koledova Z
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- Mice, Animals, Morphogenesis physiology, Coculture Techniques, Fibroblasts metabolism, Mammary Glands, Animal metabolism, Epithelial Cells metabolism
- Abstract
Epithelial branching morphogenesis is an essential process in living organisms, through which organ-specific epithelial shapes are created. Interactions between epithelial cells and their stromal microenvironment instruct branching morphogenesis but remain incompletely understood. Here, we employed fibroblast-organoid or fibroblast-spheroid co-culture systems and time-lapse imaging to reveal that physical contact between fibroblasts and epithelial cells and fibroblast contractility are required to induce mammary epithelial branching. Pharmacological inhibition of ROCK or non-muscle myosin II, or fibroblast-specific knock-out of Myh9 abrogate fibroblast-induced epithelial branching. The process of fibroblast-induced branching requires epithelial proliferation and is associated with distinctive epithelial patterning of yes associated protein (YAP) activity along organoid branches, which is dependent on fibroblast contractility. Moreover, we provide evidence for the in vivo existence of contractile fibroblasts specifically surrounding terminal end buds (TEBs) of pubertal murine mammary glands, advocating for an important role of fibroblast contractility in branching in vivo. Together, we identify fibroblast contractility as a novel stromal factor driving mammary epithelial morphogenesis. Our study contributes to comprehensive understanding of overlapping but divergent employment of mechanically active fibroblasts in developmental versus tumorigenic programs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Sumbal et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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14. Fourteenth Annual ENBDC Workshop: Methods in Mammary Gland Biology and Breast Cancer.
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Chalmers SB, van der Wal T, Fre S, and Jonkers J
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- Humans, Animals, Female, Breast pathology, Tumor Microenvironment, Biology, Mammary Glands, Animal pathology, Breast Neoplasms pathology, Mammary Glands, Human pathology
- Abstract
The fourteenth annual workshop of the European Network for Breast Development and Cancer (ENBDC) on Methods in Mammary Gland Biology and Breast Cancer was held on April 26th - 29th in Weggis, Switzerland. For the first time, early career researchers organised and took part in an additional ECR workshop on the 26th of April, which was received with great enthusiasm. The topics of the main workshop included mammary branching and morphogenesis, novel experimental systems (model organisms), systemic influences on tumour progression and the tumour microenvironment. Novel and recent findings were shared across excellent oral and poster presentations., (© 2023. Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2023
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15. Imagine beyond: recent breakthroughs and next challenges in mammary gland biology and breast cancer research.
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van Amerongen R, Bentires-Alj M, van Boxtel AL, Clarke RB, Fre S, Suarez EG, Iggo R, Jechlinger M, Jonkers J, Mikkola ML, Koledova ZS, Sørlie T, and Vivanco MD
- Subjects
- Humans, Female, Breast, Biology, Mammary Glands, Human, Breast Neoplasms
- Abstract
On 8 December 2022 the organizing committee of the European Network for Breast Development and Cancer labs (ENBDC) held its fifth annual Think Tank meeting in Amsterdam, the Netherlands. Here, we embraced the opportunity to look back to identify the most prominent breakthroughs of the past ten years and to reflect on the main challenges that lie ahead for our field in the years to come. The outcomes of these discussions are presented in this position paper, in the hope that it will serve as a summary of the current state of affairs in mammary gland biology and breast cancer research for early career researchers and other newcomers in the field, and as inspiration for scientists and clinicians to move the field forward., (© 2023. The Author(s).)
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- 2023
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16. DRAG in situ barcoding reveals an increased number of HSPCs contributing to myelopoiesis with age.
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Urbanus J, Cosgrove J, Beltman JB, Elhanati Y, Moral RA, Conrad C, van Heijst JW, Tubeuf E, Velds A, Kok L, Merle C, Magnusson JP, Guyonnet L, Frisén J, Fre S, Walczak AM, Mora T, Jacobs H, Schumacher TN, and Perié L
- Subjects
- Adult, Humans, Aged, Bone Marrow, Bone Marrow Cells, Myeloid Cells, Myelopoiesis genetics, Hematopoietic Stem Cells
- Abstract
Ageing is associated with changes in the cellular composition of the immune system. During ageing, hematopoietic stem and progenitor cells (HSPCs) that produce immune cells are thought to decline in their regenerative capacity. However, HSPC function has been mostly assessed using transplantation assays, and it remains unclear how HSPCs age in the native bone marrow niche. To address this issue, we present an in situ single cell lineage tracing technology to quantify the clonal composition and cell production of single cells in their native niche. Our results demonstrate that a pool of HSPCs with unequal output maintains myelopoiesis through overlapping waves of cell production throughout adult life. During ageing, the increased frequency of myeloid cells is explained by greater numbers of HSPCs contributing to myelopoiesis rather than the increased myeloid output of individual HSPCs. Strikingly, the myeloid output of HSPCs remains constant over time despite accumulating significant transcriptomic changes throughout adulthood. Together, these results show that, unlike emergency myelopoiesis post-transplantation, aged HSPCs in their native microenvironment do not functionally decline in their regenerative capacity., (© 2023. The Author(s).)
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- 2023
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17. Paracrine signalling between intestinal epithelial and tumour cells induces a regenerative programme.
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Jacquemin G, Wurmser A, Huyghe M, Sun W, Homayed Z, Merle C, Perkins M, Qasrawi F, Richon S, Dingli F, Arras G, Loew D, Vignjevic D, Pannequin J, and Fre S
- Subjects
- Animals, Ecosystem, Epithelial Cells metabolism, Mice, Signal Transduction, Transcription Factors metabolism, Adaptor Proteins, Signal Transducing metabolism, Neoplasms
- Abstract
Tumours are complex ecosystems composed of different types of cells that communicate and influence each other. While the critical role of stromal cells in affecting tumour growth is well established, the impact of mutant cancer cells on healthy surrounding tissues remains poorly defined. Here, using mouse intestinal organoids, we uncover a paracrine mechanism by which intestinal cancer cells reactivate foetal and regenerative YAP-associated transcriptional programmes in neighbouring wildtype epithelial cells, rendering them adapted to thrive in the tumour context. We identify the glycoprotein thrombospondin-1 (THBS1) as the essential factor that mediates non-cell-autonomous morphological and transcriptional responses. Importantly, Thbs1 is associated with bad prognosis in several human cancers. This study reveals the THBS1-YAP axis as the mechanistic link mediating paracrine interactions between epithelial cells in intestinal tumours., Competing Interests: GJ, AW, MH, WS, ZH, CM, MP, FQ, SR, FD, GA, DL, DV, JP, SF No competing interests declared, (© 2022, Jacquemin et al.)
- Published
- 2022
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18. In vivo imaging of mammary epithelial cell dynamics in response to lineage-biased Wnt/β-catenin activation.
- Author
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Lloyd-Lewis B, Gobbo F, Perkins M, Jacquemin G, Huyghe M, Faraldo MM, and Fre S
- Subjects
- Animals, Epithelial Cells metabolism, Epithelium metabolism, Hyperplasia pathology, Wnt Signaling Pathway, Mammary Glands, Animal metabolism, beta Catenin metabolism
- Abstract
Real-time in vivo imaging provides an essential window into the spatiotemporal cellular events contributing to tissue development and pathology. By coupling longitudinal intravital imaging with genetic lineage tracing, here we capture the earliest cellular events arising in response to active Wnt/β-catenin signaling and the ensuing impact on the organization and differentiation of the mammary epithelium. This enables us to interrogate how Wnt/β-catenin regulates the dynamics of distinct subpopulations of mammary epithelial cells in vivo and in real time. We show that β-catenin stabilization, when targeted to either the mammary luminal or basal epithelial lineage, leads to cellular rearrangements that precipitate the formation of hyperplastic lesions that undergo squamous transdifferentiation. These results enhance our understanding of the earliest stages of hyperplastic lesion formation in vivo and reveal that, in mammary neoplastic development, β-catenin activation dictates a hair follicle/epidermal differentiation program independently of the targeted cell of origin., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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19. Lineage Tracing Methods to Study Mammary Epithelial Hierarchies In Vivo.
- Author
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Rodilla V and Fre S
- Subjects
- Animals, Breast cytology, Cell Lineage
- Abstract
Lineage tracing is now considered the gold standard approach to study cellular hierarchies and cell fate in vivo (McKenna and Gagnon, Development 146:dev169730, 2019; Kretzschmar and Watt, Cell 148:33-45, 2012). This type of clonal analysis consists of genetically labeling defined cells and following their destiny and progeny in vivo and in situ.Here we will describe different existing in vivo systems to clonally trace targeted cells and will discuss their respective advantages and inconveniences; we will then provide stepwise instructions for setting up and evaluate lineage tracing experiments, listing the most common downstream analyses and read-out assays., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2022
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20. Longitudinal high-resolution imaging through a flexible intravital imaging window.
- Author
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Jacquemin G, Benavente-Diaz M, Djaber S, Bore A, Dangles-Marie V, Surdez D, Tajbakhsh S, Fre S, and Lloyd-Lewis B
- Abstract
Intravital microscopy (IVM) is a powerful technique that enables imaging of internal tissues at (sub)cellular resolutions in living animals. Here, we present a silicone-based imaging window consisting of a fully flexible, sutureless design that is ideally suited for long-term, longitudinal IVM of growing tissues and tumors. Crucially, we show that this window, without any customization, is suitable for numerous anatomical locations in mice using a rapid and standardized implantation procedure. This low-cost device represents a substantial technological and performance advance that facilitates intravital imaging in diverse contexts in higher organisms, opening previously unattainable avenues for in vivo imaging of soft and fragile tissues., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).)
- Published
- 2021
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21. Notch signalling: sensor and instructor of the microenvironment to coordinate cell fate and organ morphogenesis.
- Author
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Lloyd-Lewis B, Mourikis P, and Fre S
- Subjects
- Animals, Cell Differentiation genetics, Organogenesis, Stem Cell Niche, Cell Lineage, Cellular Microenvironment, Receptors, Notch metabolism, Signal Transduction
- Abstract
During development, stem cells give rise to specialised cell types in a tightly regulated, spatiotemporal manner to drive the formation of complex three-dimensional tissues. While mechanistic insights into the gene regulatory pathways that guide cell fate choices are emerging, how morphogenetic changes are coordinated with cell fate specification remains a fundamental question in organogenesis and adult tissue homeostasis. The requirement of cell contacts for Notch signalling makes it a central pathway capable of linking dynamic cellular rearrangements during tissue morphogenesis with stem cell function. Here, we highlight recent studies that support a critical role for the Notch pathway in translating microenvironmental cues into cell fate decisions, guiding the development of diverse organ systems., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Published
- 2019
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22. Lineage tracing of Notch1-expressing cells in intestinal tumours reveals a distinct population of cancer stem cells.
- Author
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Mourao L, Jacquemin G, Huyghe M, Nawrocki WJ, Menssouri N, Servant N, and Fre S
- Subjects
- Animals, Biomarkers metabolism, Cell Differentiation physiology, Cell Proliferation physiology, Cell Self Renewal physiology, Cell Transformation, Neoplastic metabolism, Colonic Neoplasms pathology, Gene Expression Profiling methods, Humans, Intestinal Mucosa metabolism, Intestinal Neoplasms pathology, Intestines pathology, Intestines physiology, Mice, Mice, Transgenic, Receptor, Notch1 metabolism, Receptors, G-Protein-Coupled genetics, Receptors, G-Protein-Coupled metabolism, Signal Transduction, Cell Lineage physiology, Neoplastic Stem Cells metabolism, Receptor, Notch1 genetics
- Abstract
Colon tumours are hierarchically organized and contain multipotent self-renewing cells, called Cancer Stem Cells (CSCs). We have previously shown that the Notch1 receptor is expressed in Intestinal Stem Cells (ISCs); given the critical role played by Notch signalling in promoting intestinal tumourigenesis, we explored Notch1 expression in tumours. Combining lineage tracing in two tumour models with transcriptomic analyses, we found that Notch1+ tumour cells are undifferentiated, proliferative and capable of indefinite self-renewal and of generating a heterogeneous clonal progeny. Molecularly, the transcriptional signature of Notch1+ tumour cells highly correlates with ISCs, suggestive of their origin from normal crypt cells. Surprisingly, Notch1+ expression labels a subset of CSCs that shows reduced levels of Lgr5, a reported CSCs marker. The existence of distinct stem cell populations within intestinal tumours highlights the necessity of better understanding their hierarchy and behaviour, to identify the correct cellular targets for therapy.
- Published
- 2019
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23. Cellular Plasticity of Mammary Epithelial Cells Underlies Heterogeneity of Breast Cancer.
- Author
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Rodilla V and Fre S
- Abstract
The hierarchical relationships between stem cells, lineage-committed progenitors, and differentiated cells remain unclear in several tissues, due to a high degree of cell plasticity, allowing cells to switch between different cell states. The mouse mammary gland, similarly to other tissues such as the prostate, the sweat gland, and the respiratory tract airways, consists of an epithelium exclusively maintained by unipotent progenitors throughout adulthood. Such unipotent progenitors, however, retain a remarkable cellular plasticity, as they can revert to multipotency during epithelial regeneration as well as upon oncogene activation. Here, we revise the current knowledge on mammary cell hierarchies in light of the most recent lineage tracing studies performed in the mammary gland and highlight how stem cell differentiation or reversion to multipotency are at the base of tumor development and progression. In addition, we will discuss the current knowledge about the interplay between tumor cells of origin and defined genetic mutations, leading to different tumor types, and its implications in choosing specific therapeutic protocols for breast cancer patients.
- Published
- 2018
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24. Clonal analysis of Notch1-expressing cells reveals the existence of unipotent stem cells that retain long-term plasticity in the embryonic mammary gland.
- Author
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Lilja AM, Rodilla V, Huyghe M, Hannezo E, Landragin C, Renaud O, Leroy O, Rulands S, Simons BD, and Fre S
- Subjects
- Adult Stem Cells metabolism, Adult Stem Cells pathology, Animals, Cell Transformation, Neoplastic genetics, Cell Transformation, Neoplastic metabolism, Cell Transformation, Neoplastic pathology, Female, Fluorescent Antibody Technique, Gene Expression Regulation, Developmental, Gestational Age, Mammary Glands, Animal embryology, Mice, Mice, Transgenic, Models, Genetic, Morphogenesis, Neoplastic Stem Cells metabolism, Neoplastic Stem Cells pathology, Phenotype, Receptor, Notch1 genetics, Signal Transduction, Single-Cell Analysis, Time Factors, Cell Differentiation, Cell Lineage, Cell Plasticity, Epithelial Cells metabolism, Mammary Glands, Animal metabolism, Mouse Embryonic Stem Cells metabolism, Receptor, Notch1 metabolism
- Abstract
Recent lineage tracing studies have revealed that mammary gland homeostasis relies on unipotent stem cells. However, whether and when lineage restriction occurs during embryonic mammary development, and which signals orchestrate cell fate specification, remain unknown. Using a combination of in vivo clonal analysis with whole mount immunofluorescence and mathematical modelling of clonal dynamics, we found that embryonic multipotent mammary cells become lineage-restricted surprisingly early in development, with evidence for unipotency as early as E12.5 and no statistically discernable bipotency after E15.5. To gain insights into the mechanisms governing the switch from multipotency to unipotency, we used gain-of-function Notch1 mice and demonstrated that Notch activation cell autonomously dictates luminal cell fate specification to both embryonic and basally committed mammary cells. These functional studies have important implications for understanding the signals underlying cell plasticity and serve to clarify how reactivation of embryonic programs in adult cells can lead to cancer.
- Published
- 2018
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25. Impaired PRC2 activity promotes transcriptional instability and favors breast tumorigenesis.
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Wassef M, Rodilla V, Teissandier A, Zeitouni B, Gruel N, Sadacca B, Irondelle M, Charruel M, Ducos B, Michaud A, Caron M, Marangoni E, Chavrier P, Le Tourneau C, Kamal M, Pasmant E, Vidaud M, Servant N, Reyal F, Meseure D, Vincent-Salomon A, Fre S, and Margueron R
- Subjects
- Animals, Animals, Genetically Modified, Breast Neoplasms diagnosis, Breast Neoplasms genetics, Cell Line, Tumor, Disease Models, Animal, Enhancer of Zeste Homolog 2 Protein, Female, Histones metabolism, Homeostasis genetics, Humans, Male, Polycomb Repressive Complex 2 genetics, Prognosis, Prostatic Neoplasms diagnosis, Prostatic Neoplasms enzymology, Prostatic Neoplasms genetics, Breast Neoplasms enzymology, Carcinogenesis genetics, Gene Expression Regulation, Neoplastic genetics, Polycomb Repressive Complex 2 metabolism
- Abstract
Alterations of chromatin modifiers are frequent in cancer, but their functional consequences often remain unclear. Focusing on the Polycomb protein EZH2 that deposits the H3K27me3 (trimethylation of Lys27 of histone H3) mark, we showed that its high expression in solid tumors is a consequence, not a cause, of tumorigenesis. In mouse and human models, EZH2 is dispensable for prostate cancer development and restrains breast tumorigenesis. High EZH2 expression in tumors results from a tight coupling to proliferation to ensure H3K27me3 homeostasis. However, this process malfunctions in breast cancer. Low EZH2 expression relative to proliferation and mutations in Polycomb genes actually indicate poor prognosis and occur in metastases. We show that while altered EZH2 activity consistently modulates a subset of its target genes, it promotes a wider transcriptional instability. Importantly, transcriptional changes that are consequences of EZH2 loss are predominantly irreversible. Our study provides an unexpected understanding of EZH2's contribution to solid tumors with important therapeutic implications., (© 2015 Wassef et al.; Published by Cold Spring Harbor Laboratory Press.)
- Published
- 2015
- Full Text
- View/download PDF
26. Mechanical induction of the tumorigenic β-catenin pathway by tumour growth pressure.
- Author
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Fernández-Sánchez ME, Barbier S, Whitehead J, Béalle G, Michel A, Latorre-Ossa H, Rey C, Fouassier L, Claperon A, Brullé L, Girard E, Servant N, Rio-Frio T, Marie H, Lesieur S, Housset C, Gennisson JL, Tanter M, Ménager C, Fre S, Robine S, and Farge E
- Subjects
- Active Transport, Cell Nucleus, Animals, Epithelial Cells cytology, Epithelial Cells pathology, Female, Gene Expression Regulation, Neoplastic, Magnets, Male, Metal Nanoparticles, Mice, Mice, Inbred C57BL, Phosphorylation, Proto-Oncogene Proteins c-ret metabolism, Receptors, Notch genetics, Receptors, Notch metabolism, Signal Transduction, beta Catenin metabolism, Carcinogenesis pathology, Colonic Neoplasms physiopathology, Pressure, Tumor Microenvironment, beta Catenin genetics
- Abstract
The tumour microenvironment may contribute to tumorigenesis owing to mechanical forces such as fibrotic stiffness or mechanical pressure caused by the expansion of hyper-proliferative cells. Here we explore the contribution of the mechanical pressure exerted by tumour growth onto non-tumorous adjacent epithelium. In the early stage of mouse colon tumour development in the Notch(+)Apc(+/1638N) mouse model, we observed mechanistic pressure stress in the non-tumorous epithelial cells caused by hyper-proliferative adjacent crypts overexpressing active Notch, which is associated with increased Ret and β-catenin signalling. We thus developed a method that allows the delivery of a defined mechanical pressure in vivo, by subcutaneously inserting a magnet close to the mouse colon. The implanted magnet generated a magnetic force on ultra-magnetic liposomes, stabilized in the mesenchymal cells of the connective tissue surrounding colonic crypts after intravenous injection. The magnetically induced pressure quantitatively mimicked the endogenous early tumour growth stress in the order of 1,200 Pa, without affecting tissue stiffness, as monitored by ultrasound strain imaging and shear wave elastography. The exertion of pressure mimicking that of tumour growth led to rapid Ret activation and downstream phosphorylation of β-catenin on Tyr654, imparing its interaction with the E-cadherin in adherens junctions, and which was followed by β-catenin nuclear translocation after 15 days. As a consequence, increased expression of β-catenin-target genes was observed at 1 month, together with crypt enlargement accompanying the formation of early tumorous aberrant crypt foci. Mechanical activation of the tumorigenic β-catenin pathway suggests unexplored modes of tumour propagation based on mechanical signalling pathways in healthy epithelial cells surrounding the tumour, which may contribute to tumour heterogeneity.
- Published
- 2015
- Full Text
- View/download PDF
27. Luminal progenitors restrict their lineage potential during mammary gland development.
- Author
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Rodilla V, Dasti A, Huyghe M, Lafkas D, Laurent C, Reyal F, and Fre S
- Subjects
- Animals, Cell Differentiation, Cell Tracking, Epithelial Cells metabolism, Estrogen Receptor alpha genetics, Estrogen Receptor alpha metabolism, Female, Gene Expression Regulation, Developmental, Gene Knock-In Techniques, Integrases genetics, Integrases metabolism, Mammary Glands, Animal growth & development, Mammary Glands, Animal metabolism, Mice, Mice, Transgenic, Multipotent Stem Cells metabolism, Neoplastic Stem Cells metabolism, Neoplastic Stem Cells pathology, Phenotype, Pregnancy, Receptor, Notch1 genetics, Receptor, Notch1 metabolism, Cell Lineage genetics, Epithelial Cells cytology, Mammary Glands, Animal cytology, Multipotent Stem Cells cytology, Organogenesis genetics
- Abstract
The hierarchical relationships between stem cells and progenitors that guide mammary gland morphogenesis are still poorly defined. While multipotent basal stem cells have been found within the myoepithelial compartment, the in vivo lineage potential of luminal progenitors is unclear. Here we used the expression of the Notch1 receptor, previously implicated in mammary gland development and tumorigenesis, to elucidate the hierarchical organization of mammary stem/progenitor cells by lineage tracing. We found that Notch1 expression identifies multipotent stem cells in the embryonic mammary bud, which progressively restrict their lineage potential during mammary ductal morphogenesis to exclusively generate an ERαneg luminal lineage postnatally. Importantly, our results show that Notch1-labelled cells represent the alveolar progenitors that expand during pregnancy and survive multiple successive involutions. This study reveals that postnatal luminal epithelial cells derive from distinct self-sustained lineages that may represent the cells of origin of different breast cancer subtypes.
- Published
- 2015
- Full Text
- View/download PDF
28. Tripartite interactions between Wnt signaling, Notch and Myb for stem/progenitor cell functions during intestinal tumorigenesis.
- Author
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Germann M, Xu H, Malaterre J, Sampurno S, Huyghe M, Cheasley D, Fre S, and Ramsay RG
- Subjects
- Adenoma metabolism, Adenoma mortality, Adenoma pathology, Animals, Carcinogenesis, Cell Differentiation, Cell Proliferation, Cells, Cultured, Intestinal Mucosa metabolism, Intestines cytology, Kaplan-Meier Estimate, Mice, Mice, Inbred C57BL, Mice, Knockout, Organoids cytology, Organoids metabolism, Receptors, G-Protein-Coupled genetics, Receptors, G-Protein-Coupled metabolism, Receptors, Notch genetics, Signal Transduction, Stem Cells cytology, Nerve Tissue Proteins metabolism, Proto-Oncogene Proteins c-myb metabolism, Receptors, Notch metabolism, Stem Cells metabolism, Wnt Proteins metabolism
- Abstract
Deletion studies confirm Wnt, Notch and Myb transcriptional pathway engagement in intestinal tumorigenesis. Nevertheless, their contrasting and combined roles when activated have not been elucidated. This is important as these pathways are not ablated but rather are aberrantly activated during carcinogenesis. Using ApcMin/+ mice as a source of organoids we documented their transition, on a clone-by-clone basis, to cyst-like spheres with constitutively activated Wnt pathway, increased self-renewal and growth and reduced differentiation. We then looked at this transition when Myb and/or Notch1 are activated. Activated Notch promoted cyst-like organoids. Conversely growth and propagation of cyst-like, but not normal organoids were Notch-independent. Activated Myb promoted normal, but not cyst-like organoids. Interestingly the Wnt, Notch and Myb pathways were all involved in regulating the expression of the intestinal stem cell (ISC) gene Lgr5 in organoids, while ISC gene and Notch target Olfm4 was dominantly repressed by Wnt. These findings parallel mouse intestinal adenoma formation where Notch promoted the initiation, but not growth, of Wnt-driven Olfm4-repressed colon tumors. Also Myb was essential for colon tumor initiation and collateral mouse pathologies. These data reveal the complex interplay and hierarchy of transcriptional networks that operate in ISCs and uncover a shift in pathway-dependencies during tumor initiation., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2014
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- View/download PDF
29. Concomitant Notch activation and p53 deletion trigger epithelial-to-mesenchymal transition and metastasis in mouse gut.
- Author
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Chanrion M, Kuperstein I, Barrière C, El Marjou F, Cohen D, Vignjevic D, Stimmer L, Paul-Gilloteaux P, Bièche I, Tavares Sdos R, Boccia GF, Cacheux W, Meseure D, Fre S, Martignetti L, Legoix-Né P, Girard E, Fetler L, Barillot E, Louvard D, Zinovyev A, and Robine S
- Subjects
- Animals, Base Sequence, Cell Lineage, DNA Primers genetics, Exome genetics, Gastrointestinal Tract metabolism, Genotype, Green Fluorescent Proteins metabolism, Humans, Image Processing, Computer-Assisted, Immunohistochemistry, Mice, Mice, Transgenic, Molecular Sequence Data, Reverse Transcriptase Polymerase Chain Reaction, Sequence Analysis, DNA, Signal Transduction genetics, Signal Transduction physiology, Disease Models, Animal, Epithelial-Mesenchymal Transition physiology, Gastrointestinal Tract physiology, Neoplasm Metastasis physiopathology, Receptor, Notch1 metabolism, Tumor Suppressor Protein p53 metabolism
- Abstract
Epithelial-to-mesenchymal transition-like (EMT-like) is a critical process allowing initiation of metastases during tumour progression. Here, to investigate its role in intestinal cancer, we combine computational network-based and experimental approaches to create a mouse model with high metastatic potential. Construction and analysis of this network map depicting molecular mechanisms of EMT regulation based on the literature suggests that Notch activation and p53 deletion have a synergistic effect in activating EMT-like processes. To confirm this prediction, we generate transgenic mice by conditionally activating the Notch1 receptor and deleting p53 in the digestive epithelium (NICD/p53(-/-)). These mice develop metastatic tumours with high penetrance. Using GFP lineage tracing, we identify single malignant cells with mesenchymal features in primary and metastatic tumours in vivo. The development of such a model that recapitulates the cellular features observed in invasive human colorectal tumours is appealing for innovative drug discovery.
- Published
- 2014
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- View/download PDF
30. Notch3 marks clonogenic mammary luminal progenitor cells in vivo.
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Lafkas D, Rodilla V, Huyghe M, Mourao L, Kiaris H, and Fre S
- Subjects
- Animals, Biomarkers metabolism, Cell Survival, Coculture Techniques, Feeder Cells, Female, Mammary Glands, Animal cytology, Mice, Mice, Inbred C57BL, Mice, Transgenic, NIH 3T3 Cells, Pregnancy, Receptor, Notch3, Receptors, Notch genetics, Signal Transduction, Time Factors, Cell Lineage, Cell Proliferation, Mammary Glands, Animal metabolism, Receptors, Notch metabolism, Stem Cells metabolism
- Abstract
The identity of mammary stem and progenitor cells remains poorly understood, mainly as a result of the lack of robust markers. The Notch signaling pathway has been implicated in mammary gland development as well as in tumorigenesis in this tissue. Elevated expression of the Notch3 receptor has been correlated to the highly aggressive "triple negative" human breast cancer. However, the specific cells expressing this Notch paralogue in the mammary gland remain unknown. Using a conditionally inducible Notch3-CreERT2(SAT) transgenic mouse, we genetically marked Notch3-expressing cells throughout mammary gland development and followed their lineage in vivo. We demonstrate that Notch3 is expressed in a highly clonogenic and transiently quiescent luminal progenitor population that gives rise to a ductal lineage. These cells are capable of surviving multiple successive pregnancies, suggesting a capacity to self-renew. Our results also uncover a role for the Notch3 receptor in restricting the proliferation and consequent clonal expansion of these cells.
- Published
- 2013
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31. Notch signaling in intestinal homeostasis across species: the cases of Drosophila, Zebrafish and the mouse.
- Author
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Fre S, Bardin A, Robine S, and Louvard D
- Subjects
- Animals, Carcinoma genetics, Carcinoma metabolism, Carcinoma pathology, Homeostasis physiology, Humans, Intestinal Mucosa metabolism, Intestinal Neoplasms genetics, Intestinal Neoplasms metabolism, Intestinal Neoplasms pathology, Receptors, Notch genetics, Receptors, Notch metabolism, Signal Transduction genetics, Signal Transduction physiology, Species Specificity, Drosophila genetics, Drosophila metabolism, Drosophila physiology, Homeostasis genetics, Intestines physiology, Mice genetics, Mice metabolism, Mice physiology, Receptors, Notch physiology, Zebrafish genetics, Zebrafish metabolism, Zebrafish physiology
- Abstract
Notch signaling has been recently shown to have a fundamental role in stem cell maintenance and control of proper homeostasis in the intestine of different species. Here, we briefly review the current literature on Notch signals in the intestine of Drosophila, Zebrafish and the mouse, and try to highlight conserved and divergent Notch functions across species. Notch signals show a remarkably conserved role in skewing cell fate choices in intestinal lineages throughout evolution. Genetic analysis demonstrates that loss of Notch signaling invariably leads to increased numbers of secretory cells and loss of enterocytes, while gain of Notch function will completely block secretory cell differentiation. Finally, we discuss the potential contribution of Notch signaling to the initiation of colorectal cancer by controlling the maintenance of the undifferentiated state of intestinal neoplastic cells and speculate on the therapeutic consequences of affecting cancer stem cells., (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Published
- 2011
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- View/download PDF
32. Notch lineages and activity in intestinal stem cells determined by a new set of knock-in mice.
- Author
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Fre S, Hannezo E, Sale S, Huyghe M, Lafkas D, Kissel H, Louvi A, Greve J, Louvard D, and Artavanis-Tsakonas S
- Subjects
- Animals, Basic Helix-Loop-Helix Transcription Factors genetics, Basic Helix-Loop-Helix Transcription Factors metabolism, Cell Differentiation, Clone Cells, Enterocytes cytology, Enterocytes metabolism, Gene Targeting, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, Integrases metabolism, Kinetics, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microvilli metabolism, Multipotent Stem Cells cytology, Multipotent Stem Cells metabolism, Sequence Homology, Amino Acid, Signal Transduction, Transcription Factor HES-1, Transcription, Genetic, Cell Lineage, Gene Knock-In Techniques, Intestines cytology, Receptors, Notch metabolism, Stem Cells cytology, Stem Cells metabolism
- Abstract
The conserved role of Notch signaling in controlling intestinal cell fate specification and homeostasis has been extensively studied. Nevertheless, the precise identity of the cells in which Notch signaling is active and the role of different Notch receptor paralogues in the intestine remain ambiguous, due to the lack of reliable tools to investigate Notch expression and function in vivo. We generated a new series of transgenic mice that allowed us, by lineage analysis, to formally prove that Notch1 and Notch2 are specifically expressed in crypt stem cells. In addition, a novel Notch reporter mouse, Hes1-EmGFP(SAT), demonstrated exclusive Notch activity in crypt stem cells and absorptive progenitors. This roster of knock-in and reporter mice represents a valuable resource to functionally explore the Notch pathway in vivo in virtually all tissues.
- Published
- 2011
- Full Text
- View/download PDF
33. Notch and Wnt signals cooperatively control cell proliferation and tumorigenesis in the intestine.
- Author
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Fre S, Pallavi SK, Huyghe M, Laé M, Janssen KP, Robine S, Artavanis-Tsakonas S, and Louvard D
- Subjects
- Adenoma genetics, Adenoma metabolism, Adenoma pathology, Animals, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors, Cell Differentiation, Cell Proliferation, Cell Transformation, Neoplastic genetics, Drosophila Proteins genetics, Drosophila Proteins metabolism, Drosophila melanogaster, Humans, Intestinal Neoplasms genetics, Mice, Mice, Inbred C57BL, Mice, Knockout, Nerve Tissue Proteins deficiency, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Receptors, Notch genetics, Survival Rate, TCF Transcription Factors deficiency, TCF Transcription Factors genetics, TCF Transcription Factors metabolism, Transcription Factor 4, Wnt1 Protein genetics, Wnt1 Protein metabolism, Cell Transformation, Neoplastic metabolism, Cell Transformation, Neoplastic pathology, Intestinal Neoplasms metabolism, Intestinal Neoplasms pathology, Receptors, Notch metabolism, Signal Transduction, Wnt Proteins metabolism
- Abstract
Notch and Wnt signals play essential roles in intestinal development and homeostasis, yet how they integrate their action to affect intestinal morphogenesis is not understood. We examined the interplay between these two signaling pathways in vivo, by modulating Notch activity in mice carrying either a loss- or a gain-of-function mutation of Wnt signaling. We find that the dramatic proliferative effect that Notch signals have on early intestinal precursors requires normal Wnt signaling, whereas its influence on intestinal differentiation appears independent of Wnt. Analogous experiments in Drosophila demonstrate that the synergistic effects of Notch and Wnt are valid across species. We also demonstrate a striking synergy between Notch and Wnt signals that results in inducing the formation of intestinal adenomas, particularly in the colon, a region rarely affected in available mouse tumor models, but the primary target organ in human patients. These studies thus reveal a previously unknown oncogenic potential of Notch signaling in colorectal tumorigenesis that, significantly, is supported by the analysis of human tumors. Importantly, our experimental evidence raises the possibility that Notch activation might be an essential initial event triggering colorectal cancer.
- Published
- 2009
- Full Text
- View/download PDF
34. Epithelial morphogenesis and intestinal cancer: new insights in signaling mechanisms.
- Author
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Fre S, Vignjevic D, Schoumacher M, Duffy SL, Janssen KP, Robine S, and Louvard D
- Subjects
- Animals, Bone Morphogenetic Proteins genetics, Bone Morphogenetic Proteins physiology, Gene Expression Regulation, Developmental, Gene Expression Regulation, Neoplastic, Hedgehog Proteins genetics, Hedgehog Proteins physiology, Humans, Intestinal Mucosa physiology, Intestinal Neoplasms pathology, Models, Biological, Morphogenesis physiology, Neoplasm Metastasis, PTEN Phosphohydrolase genetics, PTEN Phosphohydrolase physiology, Receptor Protein-Tyrosine Kinases genetics, Receptor Protein-Tyrosine Kinases physiology, Receptors, Notch genetics, Receptors, Notch physiology, Wnt Proteins genetics, Wnt Proteins physiology, beta Catenin genetics, beta Catenin physiology, Intestinal Mucosa embryology, Intestinal Neoplasms genetics, Morphogenesis genetics, Signal Transduction genetics
- Abstract
In this review, the major signal transduction pathways that have been shown to play an important role in intestinal homeostasis are highlighted. Each of them, the Wnt, Notch, Hedgehog, and Bone Morphogenetic Protein, as well as growth-factor regulated Receptor Tyrosine Kinases are depicted with a special emphasis through their involvement in stem cell maintenance and their role in intestinal tumorigenesis. Finally, we discuss recent data on the final steps of tumor progression, notably the formation of distant metastases. This multistep process is highly complex and still far from being understood while being of major importance for the survival of patients with digestive cancer.
- Published
- 2008
- Full Text
- View/download PDF
35. [Notch signals control the fate of immature progenitor cells in the intestine].
- Author
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Robine S, Fre S, Huyghe M, Artavanis-Tsakonas S, and Louvard D
- Subjects
- Animals, Intestines cytology, Signal Transduction physiology, Stem Cells cytology, Intestines physiology, Stem Cells physiology
- Published
- 2005
- Full Text
- View/download PDF
36. Notch signals control the fate of immature progenitor cells in the intestine.
- Author
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Fre S, Huyghe M, Mourikis P, Robine S, Louvard D, and Artavanis-Tsakonas S
- Subjects
- Animals, Apoptosis, Basic Helix-Loop-Helix Transcription Factors, Cell Differentiation, Cell Proliferation, DNA-Binding Proteins genetics, Epithelial Cells cytology, Epithelial Cells metabolism, Female, Gene Expression Regulation, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, Integrases genetics, Integrases metabolism, Male, Mice, Mice, Transgenic, Nerve Tissue Proteins genetics, RNA, Messenger genetics, RNA, Messenger metabolism, Receptor, Notch1, Receptors, Cell Surface genetics, Transcription Factor HES-1, Transcription Factors genetics, Viral Proteins genetics, Viral Proteins metabolism, Cell Lineage, Intestinal Mucosa metabolism, Intestines cytology, Receptors, Cell Surface metabolism, Signal Transduction, Stem Cells cytology, Stem Cells metabolism, Transcription Factors metabolism
- Abstract
The Notch signalling pathway plays a crucial role in specifying cellular fates in metazoan development by regulating communication between adjacent cells. Correlative studies suggested an involvement of Notch in intestinal development. Here, by modulating Notch activity in the mouse intestine, we directly implicate Notch signals in intestinal cell lineage specification. We also show that Notch activation is capable of amplifying the intestinal progenitor pool while inhibiting cell differentiation. We conclude that Notch activity is required for the maintenance of proliferating crypt cells in the intestinal epithelium.
- Published
- 2005
- Full Text
- View/download PDF
37. Recognition specificity of individual EH domains of mammals and yeast.
- Author
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Paoluzi S, Castagnoli L, Lauro I, Salcini AE, Coda L, Fre' S, Confalonieri S, Pelicci PG, Di Fiore PP, and Cesareni G
- Subjects
- Adaptor Proteins, Signal Transducing, Amino Acid Sequence, Animals, Base Sequence, Binding Sites, Calcium-Binding Proteins chemistry, DNA Primers, Humans, Intracellular Signaling Peptides and Proteins, Molecular Sequence Data, Peptides metabolism, Phosphoproteins chemistry, Recombinant Proteins chemistry, Recombinant Proteins metabolism, Sequence Homology, Amino Acid, Calcium-Binding Proteins metabolism, Phosphoproteins metabolism, Saccharomyces cerevisiae metabolism
- Abstract
The Eps homology (EH) domain is a recently described protein binding module that is found, in multiple or single copies, in several proteins in species as diverse as human and yeast. In this work, we have investigated the molecular details of recognition specificity mediated by this domain family by characterizing the peptide-binding preference of 11 different EH domains from mammal and yeast proteins. Ten of the eleven EH domains could bind at least some peptides containing an Asn-Pro-Phe (NPF) motif. By contrast, the first EH domain of End3p preferentially binds peptides containing an His-Thr/Ser-Phe (HT/SF) motif. Domains that have a low affinity for the majority of NPF peptides reveal some affinity for a third class of peptides that contains two consecutive amino acids with aromatic side chains (FW or WW). This is the case for the third EH domain of Eps15 and for the two N-terminal domains of YBL47c. The consensus sequences derived from the peptides selected from phage-displayed peptide libraries allows for grouping of EH domains into families that are characterized by different NPF-context preference. Finally, comparison of the primary sequence of EH domains with similar or divergent specificity identifies a residue at position +3 following a conserved tryptophan, whose chemical characteristics modulate binding preference.
- Published
- 1998
- Full Text
- View/download PDF
38. Epsin is an EH-domain-binding protein implicated in clathrin-mediated endocytosis.
- Author
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Chen H, Fre S, Slepnev VI, Capua MR, Takei K, Butler MH, Di Fiore PP, and De Camilli P
- Subjects
- Adaptor Protein Complex alpha Subunits, Adaptor Proteins, Signal Transducing, Adaptor Proteins, Vesicular Transport, Amino Acid Sequence, Animals, Blotting, Northern, Brain metabolism, CHO Cells, Calcium-Binding Proteins metabolism, Carrier Proteins chemistry, Cricetinae, Membrane Proteins metabolism, Molecular Sequence Data, Neuropeptides chemistry, Phosphoproteins metabolism, Protein Binding, Rats, Recombinant Fusion Proteins metabolism, Transfection, Calcium-Binding Proteins physiology, Carrier Proteins physiology, Clathrin physiology, Endocytosis physiology, Neuropeptides physiology, Phosphoproteins physiology, Vesicular Transport Proteins
- Abstract
During endocytosis, clathrin and the clathrin adaptor protein AP-2, assisted by a variety of accessory factors, help to generate an invaginated bud at the cell membrane. One of these factors is Eps15, a clathrin-coat-associated protein that binds the alpha-adaptin subunit of AP-2. Here we investigate the function of Eps15 by characterizing an important binding partner for its region containing EH domains; this protein, epsin, is closely related to the Xenopus mitotic phosphoprotein MP90 and has a ubiquitous tissue distribution. It is concentrated together with Eps15 in presynaptic nerve terminals, which are sites specialized for the clathrin-mediated endocytosis of synaptic vesicles. The central region of epsin binds AP-2 and its carboxy-terminal region binds Eps15. Epsin is associated with clathrin coats in situ, can be co-precipitated with AP-2 and Eps15 from brain extracts, but does not co-purify with clathrin coat components in a clathrin-coated vesicle fraction. When epsin function is disrupted, clathrin-mediated endocytosis is blocked. We propose that epsin may participate, together with Eps15, in the molecular rearrangement of the clathrin coats that are required for coated-pit invagination and vesicle fission.
- Published
- 1998
- Full Text
- View/download PDF
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