Développement Normal et Pathologique des Mélanocytes

Publications de l’équipe

Année de publication : 2005

Patrick Pla, Christophe Alberti, Olga Solov'eva, Manijeh Pasdar, Takahiro Kunisada, Lionel Larue (2005 May 17)

Ednrb2 orients cell migration towards the dorsolateral neural crest pathway and promotes melanocyte differentiation.

Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society : 181-7 En savoir plus
Résumé

Endothelin receptors B (Ednrb) are involved in the development of the enteric and melanocytic lineages, which originate from neural crest cells (NCCs). In mice, trunk NCCs and their derivatives express only one Ednrb. In quail, trunk NCCs express two Ednrb: Ednrb and Ednrb2. Quail Ednrb is expressed in NCCs migrating along the ventral pathway, which gives rise to the peripheral nervous system, including enteric ganglia. Ednrb2 is upregulated in NCCs before these cells enter the dorsolateral pathway. The NCCs migrating along the dorsolateral pathway are melanocyte precursors. We analyzed the in vitro differentiation and in ovo migration of mouse embryonic stem (ES) cells expressing and not expressing Ednrb2. We generated a series of transfected ES cell lines expressing Ednrb2. This receptor, like Ednrb, oriented genuine ES cells towards melanocyte lineage differentiation in vitro. The in ovo migration of Ednrb2-expressing ES cells was massively oriented towards the dorsolateral pathway, unlike that of WT or Ednrb-expressing ES cells. Thus, Ednrb2 is involved in melanoblast differentiation and migration.

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Suzanne Carreira, Jane Goodall, Isil Aksan, S Anna La Rocca, Marie-Dominique Galibert, Laurence Denat, Lionel Larue, Colin R Goding (2005 Feb 18)

Mitf cooperates with Rb1 and activates p21Cip1 expression to regulate cell cycle progression.

Nature : 764-9 En savoir plus
Résumé

The controls that enable melanoblasts and melanoma cells to proliferate are likely to be related, but so far no key regulator of cell cycle progression specific to the melanocyte lineage has been identified. The microphthalmia-associated transcription factor Mitf has a crucial but poorly defined role in melanoblast and melanocyte survival and in differentiation. Here we show that Mitf can act as a novel anti-proliferative transcription factor able to induce a G1 cell-cycle arrest that is dependent on Mitf-mediated activation of the p21(Cip1) (CDKN1A) cyclin-dependent kinase inhibitor gene. Moreover, cooperation between Mitf and the retinoblastoma protein Rb1 potentiates the ability of Mitf to activate transcription. The results indicate that Mitf-mediated activation of p21Cip1 expression and consequent hypophosphorylation of Rb1 will contribute to cell cycle exit and activation of the differentiation programme. The mutation of genes associated with melanoma, such as INK4a or BRAF that would affect either Mitf cooperation with Rb1 or Mitf stability respectively, would impair Mitf-mediated cell cycle control.

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Année de publication : 2004

Robert Moore, Delphine Champeval, Laurence Denat, Seong-Seng Tan, Florence Faure, Sylvia Julien-Grille, Lionel Larue (2004 Jul 27)

Involvement of cadherins 7 and 20 in mouse embryogenesis and melanocyte transformation.

Oncogene : 6726-35 En savoir plus
Résumé

We have determined the expression profiles of cdh7, and the related cdh20 during development. Both transcripts are found in the adult brain, but only cadherin-20 mRNA was detected during embryogenesis. In mouse embryos, cadherin-20 is synthesized by the forebrain, anterior neural ridge, developing visual system, primitive external granular layer of the cerebellum and a subset of neural crest cells likely to develop into melanoblasts. We found that the other embryonic tissues in which cadherin-20 was synthesized depended on genetic background. Melanoma cell lines contained transcripts for cadherin-7 but not for cadherin-20. The majority of the malignant melanoma cell lines produced N-cadherin (N-Cad) and/or cadherin-7 whereas melanocyte cell lines did not. The converse was observed for E-cadherin (E-Cad). Our data suggest that during development cadherin-20 is a key player in compartmentalization of the neural tube and establishment of neural circuitry. Finally, during oncogenesis, cadherin-7, N-cad and E-cad could be used as an efficient marker set for melanoma.

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Jane Goodall, Silvia Martinozzi, Timothy J Dexter, Delphine Champeval, Suzanne Carreira, Lionel Larue, Colin R Goding (2004 Mar 17)

Brn-2 expression controls melanoma proliferation and is directly regulated by beta-catenin.

Molecular and cellular biology : 2915-22 En savoir plus
Résumé

Constitutive activation of the Wnt/beta-catenin signaling pathway is a notable feature of a large minority of cases of malignant melanoma, an aggressive and increasingly common cancer. The identification of target genes downstream from this pathway is therefore crucial to our understanding of the disease. The POU domain transcription factor Brn-2 has been implicated in control of proliferation and melanoma survival, and its expression is strongly upregulated in melanoma. We show here that in vivo Brn-2 is expressed in melanocytes but not in embryonic day 11.5 melanoblasts and that its expression is directly controlled by the Wnt/beta-catenin signaling pathway in melanoma cell lines and in transgenic mice. Moreover, silent interfering RNA-mediated inhibition of Brn-2 expression in melanoma cells overexpressing beta-catenin results in significantly decreased proliferation. These results, together with the observation that BRAF signaling also induces Brn-2 expression, reveal that Brn-2 is a focus for the convergence of two key melanoma-associated signaling pathways that are linked to cell proliferation.

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Patrick Pla, Olga Solov'eva, Robert Moore, Christophe Alberti, Takahiro Kunisada, Lionel Larue (2004 Mar 16)

Dct::lacZ ES cells: a novel cellular model to study melanocyte determination and differentiation.

Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society : 142-9 En savoir plus
Résumé

Embryonic stem (ES) cells differentiate into various cell lineages in vitro. A procedure was previously designed to promote the differentiation of ES cells towards the melanocyte lineage and to obtain large and reproducible amounts of melanocytes. To elucidate the main events that lead to the development of melanocytes in vitro, we used transgenic Dct::lacZ mouse blastocysts to establish ES cell lines expressing the lacZ reporter gene under the control of the Dct promoter. Dct, a melanoblast marker, is expressed just after melanoblast determination in vivo. We evaluated the importance of recruitment, proliferation and differentiation during melanocyte ontogeny after the in vitro differentiation of Dct::lacZ ES cells into melanocytes. We showed that bFGF and cholera toxin induce precocious melanoblast determination, associated with early melanocyte differentiation. Edn3 induced melanoblast proliferation and long-term melanoblast recruitment, but not precocious determination. The lack of basic Fibroblast Growth Factor (bFGF) and cholera toxin can be partially compensated by Edn3. Thus, Dct::lacZ ES cells can be used as a model to study determination, proliferation and differentiation in the melanocyte lineage in vitro.

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Année de publication : 2003

Patrick Pla, Lionel Larue (2003 Aug 5)

Involvement of endothelin receptors in normal and pathological development of neural crest cells.

The International journal of developmental biology : 315-25 En savoir plus
Résumé

Endothelin receptors (Ednr) are G-protein-coupled receptors with seven membrane-spanning domains and are involved in various physiological processes in adults. We review here the function of these receptors during the development and transformation of the neural crest cell-specific lineage. Neural crest cells (NCC) may be classified according to their location in the body. In particular, there are clear differences between the neural crest cells arising from the cephalic part of the embryo and those arising from the vagal and truncal part. The development of cranial and cardiac NCC requires the endothelin-1/Ednra system to be fully functional whereas the development of more posterior NCC requires full functionality of the endothelin-3/Ednrb system. Mutations have been found in the genes corresponding to these systems in mammals. These mutations principally impair pigmentation and enteric ganglia development. The precise patterns of expression of these receptors and their ligands have been determined in avian and mammalian models. Data obtained in vitro and in vivo have provided insight into the roles of these proteins in cell proliferation, migration, differentiation and transformation.

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Lionel Larue, Mayuko Kumasaka, Colin R Goding (2003 May 20)

Beta-catenin in the melanocyte lineage.

Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society : 312-7 En savoir plus
Résumé

beta-Catenin is a multifunctional protein involved in cell-cell adhesion, intracellular signalling and gene transcription. It has been implicated in the development of various lineages, including neural crest derivatives. Melanocytes are derived from neural crest cells and beta-catenin is expressed throughout the development of this cell lineage. The multifunctional activity of beta-catenin is directly associated with its participation in multi protein-protein interactions. The cell-cell adhesion function of beta-catenin is mediated by the large cadherin cell adhesion molecule family, the intracellular signalling function by its interaction with GSK3beta, and the gene transcription activity by the four known LEF/TCF DNA binding-proteins. Here, we review the known beta-catenin interacting factors and targets involved in the development and transformation of melanocytes and in particular its role in the expression of the crucial gene of melanocyte development, Mitf.

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Sylvia Julien Grille, Alfonso Bellacosa, John Upson, Andres J Klein-Szanto, Frans van Roy, Whaseon Lee-Kwon, Mark Donowitz, Philip N Tsichlis, Lionel Larue (2003 May 3)

The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines.

Cancer research : 2172-8 En savoir plus
Résumé

Epithelial-mesenchymal transition (EMT) is an important process during development and oncogenesis by which epithelial cells acquire fibroblast-like properties and show reduced intercellular adhesion and increased motility. Squamous cell carcinoma lines engineered to express constitutively active Akt underwent EMT, characterized by down-regulation of the epithelial markers desmoplakin, E-cadherin, and beta-catenin and up-regulation of the mesenchymal marker vimentin. The cells lost epithelial cell morphology and acquired fibroblast-like properties. Additionally, E-cadherin was down-regulated transcriptionally. The cells expressing constitutively active Akt exhibited reduced cell-cell adhesion, increased motility on fibronectin-coated surfaces, and increased invasiveness in animals. AKT is activated in many human carcinomas, and the AKT-driven EMT may confer the motility required for tissue invasion and metastasis. These findings suggest that future therapies based on AKT inhibition may complement conventional treatments by controlling tumor cell invasion and metastasis.

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Année de publication : 2001

P Pla, R Moore, O G Morali, S Grille, S Martinozzi, V Delmas, L Larue (2001 Oct 13)

Cadherins in neural crest cell development and transformation.

Journal of cellular physiology : 121-32 En savoir plus
Résumé

Cadherins constitute a superfamily of cell adhesion molecules involved in cell-cell interaction, histogenesis and cellular transformation. They have been implicated in the development of various lineages, including derivatives of the neural crest. Neural crest cells (NCC) emerge from the dorsal part of the neural tube after an epithelio-mesenchymal transition (EMT) and migrate through the embryo. After homing and differentiation, NCC give rise to many cell types, such as neurons, Schwann cells and melanocytes. During these steps, the pattern of expression of the various cadherins studied is very dynamic. Cadherins also display plasticity of expression during the transformation of neural crest cell derivatives. Here, we review the pattern of expression and the role of the main cadherins involved in the development and transformation of neural crest cell derivatives.

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O G Morali, V Delmas, R Moore, C Jeanney, J P Thiery, L Larue (2001 Aug 30)

IGF-II induces rapid beta-catenin relocation to the nucleus during epithelium to mesenchyme transition.

Oncogene : 4942-50 En savoir plus
Résumé

The epithelium to mesenchyme transition is thought to play a fundamental role during embryonic development and tumor progression. Loss of cell-cell adhesion and modification of both cell morphology and gene expression are the main events associated with this transition. There is a large amount of evidence suggesting that growth factors can initiate these events. Yet, the connection from growth factor induction to changes in cell adhesion and morphology is largely unknown. To elucidate this connection, we have investigated the action of IGF-II on E-cadherin/beta-catenin complex-mediated cell-cell adhesion and on beta-catenin/TCF-3 mediated gene expression. We can show that (1) IGF-II induces a rapid epithelium to mesenchymal transition; (2) IGF1R, the receptor for IGF-II, belongs to the same membrane complex as E-cadherin and beta-catenin; (3) IGF-II induces a redistribution of beta-catenin from the plasma membrane to the nucleus and an intracellular sequestration and degradation of E-cadherin; (4) IGF-II induces the transcription of beta-catenin/TCF-3 target genes. Based on the given case of IGF-II and E-cadherin/beta-catenin complex, this study reveals the backbone of a cascade connecting growth factor signaling with cell-cell adhesion during EMT.

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Année de publication : 2000

O G Morali, A Jouneau, K J McLaughlin, J P Thiery, L Larue (2000 Nov 15)

IGF-II promotes mesoderm formation.

Developmental biology : 133-45 En savoir plus
Résumé

IGF-II is abundant in the nascent mesoderm of the gastrulating mouse embryo. Its function at this developmental stage is unknown. We investigated it by following the in vitro and in vivo differentiation of several androgenetic, biparental, parthenogenetic, and androgenetic Igf2 -/- murine ES cell lines; these cells differed in endogenous IGF-II levels because Igf2 is paternally expressed in the mouse embryo in most tissues. The expression of mesoderm markers and the subsequent formation of muscle structures were correlated with endogenous IGF-II level during teratoma formation and during in vitro differentiation. In addition, the absence of Igf2 in androgenetic Igf2 -/- ES cells led to a severe impairment of mesoderm development, demonstrating the dependence of the preferential mesoderm development of androgenetic ES cells upon Igf2 activity, among the numerous known imprinted genes. The addition of exogenous IGF-II to in vitro differentiation culture medium led to a specific increase in the expression of mesoderm markers. Thus, we propose a novel model in which the binding of IGF-II to its principal signaling receptor, IGF1R, at the surface of mesoderm precursor cells increases the formation of mesoderm cells.

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A Jouneau, Y Q Yu, M Pasdar, L Larue (2000 Aug 22)

Plasticity of cadherin-catenin expression in the melanocyte lineage.

Pigment cell research / sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society : 260-72 En savoir plus
Résumé

Cadherins are calcium-dependent cell adhesion receptors with strong morphoregulatory functions. To mediate functional adhesion, cadherins must interact with actin cytoskeleton. Catenins are cytoplasmic proteins that mediate the interactions between cadherins and the cytoskeleton. In addition to their role in cell-cell adhesion, catenins also participate in signaling pathways that regulate cell growth and differentiation. Cadherins and catenins appear to be involved in melanocyte development and transformation. Here, we investigated the function of cadherin-catenin complexes in the normal development and transformation of melanocytes by studying the patterns of expression of the cell-cell adhesion molecules, E-, N- and P-cadherin, and the expression of their cytoplasmic partners, alpha-, beta- and gamma-catenin during murine development. Similar analyses were performed in vitro using murine melanoblast, melanocyte, and melanoma cell lines in the presence and absence of keratinocytes, the cells with which melanocytes interact in vivo. Overall, the results suggest that the expression of cadherins and catenins is very plastic and depends on their environment as well as the transformation status of the cells. This plasticity is important in fundamental cellular mechanisms associated with normal and pathological ontogenesis, as well as with tumorigenesis.

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V Delmas, P Pla, H Feracci, J P Thiery, R Kemler, L Larue (2000 Jan 22)

Expression of the cytoplasmic domain of E-cadherin induces precocious mammary epithelial alveolar formation and affects cell polarity and cell-matrix integrity.

Developmental biology : 491-506 En savoir plus
Résumé

Cadherins are cell adhesion molecules involved in cell-cell adhesion, signalling, and cellular proliferation and differentiation. E-cadherin is required for the formation of epithelium in vivo. We investigated the contribution of the cytoplasmic domain of E-cadherin to adhesion, signalling, and differentiation during murine mammary gland development, by in vivo expression of a gene encoding a truncated form of E-cadherin lacking the extracellular domain. The expression of this gene in mammary epithelial cells during pregnancy induced precocious lobular epithelial morphogenesis associated with morphological differentiation and the early synthesis of various molecules (advanced milk fat globule appearance and milk protein production). After delivery, when a fully differentiated and secretory epithelium is required for lactation, the cytoplasmic domain of E-cadherin had a dominant-negative effect on cell-cell adhesion and affected the structure and function of the epithelium. This also led to the partial loss of epithelial polarisation and changes in the basement membrane, both important in malignancy. Thus, the cytoplasmic domain of E-cadherin induces epithelial morphogenesis, but also alters the cohesiveness of the fully differentiated epithelium.

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Année de publication : 1999

A Beauvais-Jouneau, P Pla, F Bernex, S Dufour, J Salamero, R Fässler, J J Panthier, J P Thiery, L Larue (1999 Nov 24)

A novel model to study the dorsolateral migration of melanoblasts.

Mechanisms of development : 3-14 En savoir plus
Résumé

Melanocytes derived from pluripotent neural crest cells migrate initially in the dorsolateral pathway between the ectoderm and dermomyotome. To understand the role of specific proteins involved in this cell migration, we looked for a cellular model that mimics the in vivo behavior of melanoblasts, and that allows functional studies of their migration. We report here that wild-type embryonic stem (ES) cells are able to follow the ventral and dorsolateral neural crest pathways after being grafted into chicken embryos. By contrast, a mutant ES cell line deficient for beta1 integrin subunits, proteins involved in cell-extracellular interactions, had a severely impaired migratory behavior. Interestingly, ES cells deficient for Kit, the tyrosine kinase receptor for the stem cell factor (SCF), behaved similarly to wild-type ES cells. Thus, grafting mouse ES cells into chicken embryos provides a new cellular system that allows both in vitro and in vivo studies of the molecular mechanisms controlling dorsolateral migration.

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Année de publication : 1998

C Bonneton, L Larue, J P Thiery (1998 Apr 16)

[Current data on the role of APC protein in the origin of colorectal cancer].

Bulletin du cancer : 1053-60 En savoir plus
Résumé

The adenomatous polyposis coli (APC) gene has been found to be mutated during the development of sporadic colorectal cancers as well as in familial adenomatous polyposis (FAP). These conditions result from initially somatic and germ line mutations respectively. In both cases, the expressed protein is truncated at its carboxyterminal region. Investigations into the role of wild-type APC have led to a better understanding of the importance of mutations in the genesis and progression of adenomas. APC was shown to regulate cell growth and cell death, to bind beta-catenin, and to colocalize with microtubules. APC truncation was therefore hypothesized to alter cell multiplication and cells are no longer able to undergo apoptosis. Owing to its beta-catenin binding, APC can modify the pool of beta-catenin which is in part utilized in the assembly of adherens junctions and in nuclear signalling. Truncated APC is unable to regulate this pool thereby altering adhesion and cell signalling. Finally, APC involvement in microtubule-dependent locomotion may explain some changes in cell movement which are observed in adenomas. The establishment of murine mutants and of normal and malignant intestinal cell cultures have allowed to assess biochemical and physiological properties of APC and its putative role in the genesis of colorectal carcinogenesis. Moreover, these experimental models have suggested a variety of possible therapeutic approaches.

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