Basic Research Team 6 « Genetics and physiopathology of eptihelial cancers » Dr Thierry MAGNALDO, DR2 CNRS / Guerrino MENEGUZZI, DR1 INSERM, Team Leaders Among the epithelial cancers that arise with ageing, squamous cell carcinomas are by far the most common (figure 1) and are generally lethal. Invasiveness of epithelial cancers is known to depend on the interactions between the tumor cells and their microenvironment.. However, the exact nature of such interactions deserves further investigations. Pathogenesis of human skin carcinomas is subject to genetic, environmental, and age-related parameters. Our experimental settings aim at documenting molecular events associated with epithelial carcinogenesis and thus contribute to innovative therapeutic approaches. We set up experimental models of susceptibility to skin cancer based on rare genetic conditions such as the Gorlin’ or the Xerodema pigmentosum (XP) syndromes that are due to distinct molecular alterations (Nucleotide Excision Repair and Sonic Hedgehog signaling pathway, respectively). Most notably, Gorlin’syndrome is associated with non metastatic skin carcinoma while rates of metastatic cancers are very elevated in XP patients. Development of pathological organotypic skin cultures in vitro indicate thatstroma-epithelium interactions are affected in a disease specific manner and suggests that patients’ fibroblasts may play a key role in early tumor development and determine aggressiveness (figure 2). Our models will contribute to deciphering molecular clues underlying stroma-cancer evolution and improve targetting of antitumoral treatments.
Figure 1 adapted from de DePihno, R., Nature, 408, (2000)The age of Cancer.
Pathologic organotypic skin cultures (OSC), developed from cells isolated from patients suffering either from the cancer-susceptibility genetic diseases xeroderma pigmentousm, a, or Gorlin’ syndrome, b. Epithelial cancers - Epithelial cancers and their invasive capacity are known to depend on interaction between the tumor cell and it microenvironment but the exact nature of these interactions deserves further investigations. - In the human, skin is the organ most often victim of carcinoma. As for all carcinoma, age is a major risk factor. Skin thus constitutes a prototype model for studying epithelio-mesenchymal interactions upon aging and carcinogenesis. - Study of cells isolated from patients with the rare cancer prone genetic diseases, Xerodema pigmentosum (defect of nucleotide excision repair) or Gorlin’syndrome (defect of Hedgehog signaling due to PATCHED mutation) indicated that both epithelial and mesenchymal cells were affected in absence of genotoxic stress or external stimuli. - Notably, in organotypic skin cultures, primary patients keratinocytes exhibited tumor-like features of differenciation and invasiveness that were further enhanced in the presence of autologous fibroblasts. Patient fibroblasts overexpressed factors associated to extracellular matrix modifications (MMP1, MMP3), cell proliferation (KGF/FGF7), or inflammation (SDF1a/CXCL12). - Patients fibroblasts thus spontaneously present characteristics of stromal cells called carcinoma associated fibroblasts (CAF). Patients fibroblasts could thus play a key role in early tumor development and determine agressiveness. Deciphering molecular pathways involved in epithelial cancer development within its stromal context will contribute to better targetting antitumoral treatments. Research Team Clinical Researchers Patents Publications Warrick, E., Garcia, M., Chagnoleau, C., Chevallier, O., Bergoglio, V., Sartori, D., Mavilio, F., Angulo, J.F., Avril, M.F., Sarasin, A.., Larcher, F., Del Rio, M., Bernerd, F., Magnaldo, T. Preclinical corrective gene transfer in xeroderma pigmentosum human skin stem cells. Mol Ther. 2011 Nov 8. doi: 10.1038/mt.2011.233. [Epub ahead of print] Villeneuve, C., Baricault, L., Canelle, L., Barboule, N., Racca, C., Monsarrat, B., Magnaldo, T., Larminat, F. (2010) Mitochondrial proteomic approach reveals GAL7 as a novel BCL2 binding protein in human cells. Mol Biol Cell. 2011 Apr;22(7):999-1013. Valin, A., Avril, MF., Magnaldo, T. Dermal fibroblasts exert a key influence in the development of basal-cell skin cancers: the model of Gorlin syndrome. (2010) Med Sci 26:22-25. Valin, A., Barnay-Verdier, S., Robert, T., Ripoche, H., Brellier, F., Chevallier-Lagente, O., Avril, M. F. and Magnaldo, T. (2009). PTCH +/- dermal fibroblasts isolated from healthy skin biopsy of Gorlin syndrome patients exhibit features of carcinoma associated fibroblasts. Plos One. 4(3):e4818. Gendronneau, G., Sukhvinder S. Sidhu, S., Delacour, D., Dang, T., Calonne C., Houzelstein, D., Magnaldo, T., and Poirier, F. Galectin-7 in the control of epidermal homeostasis after injury. Mol. Biol. Cell, 2008 Oct 1. Brellier, F., Bergoglio, V., Valin, A., Barnay, S., Chevallier-Lagente, O., Viel, P., Spatz, A., Gorry, P., Avril, M. and Magnaldo, T. (2008). Heterozygous mutations in the tumor suppressor gene PATCHED provoke basal cell carcinoma-like features in human organotypic skin cultures Oncogene, (04 Aug 2008), doi: 10.1038/onc.2008.260. Magnaldo, T. (2004). Xeroderma pigmentosum: from genetics to hopes and realities of cutaneous gene therapy. Expert Opin Biol Ther 4, 169-179. Gache Y, Baldeschi C, Del Rio M, Gagnoux-Palacios L, Larcher F, Lacour JP, Meneguzzi G. (2004) "Construction of skin equivalents for gene therapy of recessive dystrophic epidermolysis bullosa". Hum Gen Ther. Oct;15(10):921-33. Gache Y., Charlesworth A., Philipe N., Vabres P., Meneguzzi G. (2004) "Identification de formes transitoires d’épidermolyses bulleuses avec atrésie pylorique". J . Invest. Dermatol. 123 :P25 Capt A., Spirito F., Guaguere E., Spadafora A., Ortonne J-P. and Meneguzzi G. (2005) "Inherited junctional epidermolysis bullosa in the German pointer: establishment of a large animal model for the condition". J Invest Dermatol 124:530-5. Gagnoux-Palacios L., Hervouet C., Spirito F., Roques S., Mezzina M., Danos O., Meneguzzi G. (2005) "Assessment of optimal transduction of primary human skin keratinocytes by viral vectors". J Gene Med. J Gene Med. 7: 1178-1186. Baudoin C., Fantin L. and Meneguzzi G. (2005) "Proteolytic processing of the laminin 3 G domain mediates assembly of hemidesmosomes but has no role on keratinocyte migration. J Invest Dermatol. 125(5):883-8. Fitsialos, AA. Chassot, L. Turchi, M.A. Dayem, K. LeBrigand, C. Moreilhon, G. Meneguzzi , R. Busca, B. Mari, P. Barbry and G. Ponzio. (2007) Transcriptional signature of epidermal keratinocytes subjected to in vitro SCRATCH wounding, reveals selective roles for ERK1/2, p38 and PI3K signalling pathways J. Biol. Chem 282, p15090-102. A.A. Chassot, L. Turchi, T. Virolle, G. Fitsialos, M. Batoz, M. Deckert, V. Dulic, G. Meneguzzi, R. Busca and G. Ponzio. (2007) Id3 is a novel regulator of p27kip1 mRNA in early G1 phase and is required for cell cycle progression. Oncogene 26, p5772-83. M D Alessio, G Zambruno, A Charlesworth, J-P Lacour, G. Meneguzzi (2008) "Immunofluorescence analysis of villous trophoblasts: a new tool for early prenatal diagnosis of inherited epidermolysis bullosa with pyloric atresia". J Invest Dermatol.128(12):2815-9. G Fitsialos, I Bourget, S Augier, A Ginouves, R Rezzonico, T Odorisio, F Cianfarani, T Virolle, J Pouyssegur, G. Meneguzzi, E Berra, G Ponzio, R Busca. (2008) HIF1 transcription factor regulates laminin-332 expression and keratinocytes migration . J Cell Sci. 15;121 (Pt 18):2992-3001. Baby Team Cédric Gaggioli " Tumor-Stroma Crosstalk in Invasive Cancer " Uncovering the signaling molecules secreted by pro-invasive “activated” fibroblasts that leads to SCC cells invasion We aim at identifying matrix proteins differentially assemble within the matrix by stimulated HDF cells compare to their non-stimulated counterpart. A comparative proteomic approach will be used to qualitatively and quantitatively identify matrix proteins assembled within the matrix of stimulated HDF cells compared to the non-stimulated counterpart. Matrix molecules layered down by activated HDF cells to culture Petri dishes will be analyzed and identified using a mass spectrometry approach. The possible role that matrix components, selected on the basis of data published in the literature, may have on SCC cell behaviour will then be assessed by conventional cell biology procedures, which include cell adhesion, migration and invasion in three dimensional organotypic assays. This approach is expected to lead to the identification of secreted molecules involved in SCC cell invasion induced by CAF. The proposed research projects aim at address fundamental aspects on invasive carcinoma development with a particular attention on the stroma as therapeutic target. Besides this contribution this project aims at identifying potential clinical target in a long-term goal for developing new strategies to counteract cancer cell invasion and metastasis. Research Team Publications Sanz-Moreno V†, Gaggioli C†, Yeo M, Albrengues J, Wallberg F, Viros A, Hooper S, Mitter R, Féral CC, Cook M, Larkin J, Marais R, Meneguzzi G, Sahai E, Marshall CJ. ROCK and JAK1 Signaling Cooperate to Control Actomyosin Contractility in Tumor Cells and Stroma. Cancer Cell. 2011 Aug 16;20(2):229-45. †Contributed equally to this work. Hooper S, Gaggioli C, Sahai E. A chemical biology screen reveals a role for Rab21-mediated control of actomyosin contractility in fibroblast-driven cancer invasion. Br J Cancer. 2010 Jan 19;102(2):392-402. Medjkane, S., Perez-Sanchez, C., Gaggioli, C, Sahai, E., and Treisman, R. Myocardin-Related Transcription Factors and SRF are required for cytoskeletal dynamics, invasion and experimental metastasis. Nature Cell Biology. 2009 Mar;11(3):257-68. Gaggioli C. Collective invasion of carcinoma cells: when the fibroblasts take the lead. Cell Adh Migr. 2008 Jan-Mar;2(1) :45-7. Gaggioli C., Hooper H., Hidalgo-Carcedo C., Grosse R., Marshall JF., Harrington K., and Sahai E. Fibroblast led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells. Nature Cell Biology, 2007 Dec;9(12):1392-400. Gaggioli C. and Sahai E. Melanoma Invasion: Current knowledge and future directions. Pigment Cell Res. 2007 Jun;20 (3):161-72. Review. Gaggioli C., Robert G., Bertolotto C., Bailet O., Abbe P., Spadafora A, Bahadoran P, Ortonne J-P, Baron V, Ballotti R. & Tartare-Deckert, S. Tumor-Derived Fibronectin Is Involved in Melanoma Cell Invasion and Regulated by V600E B-Raf Signaling Pathway. J Invest Dermatol. 2007 Feb;127(2):400-10. Guillaume Robert, Cédric Gaggioli, Carine Chavey, Olivier Bailet, Patricia Abbe, Edith Aberdam, Sabatie Emillie, Amparo Cano, Garcia De Herreros, Robert Ballotti and Sophie Tartare-Deckert. SPARC represses E-cadherin expression and induces mesenchymal transition during melanoma development. Cancer Research 2006 Aug 1;66(15):7516-23. Roser Buscà, Edurne Berra, Cédric Gaggioli, Barbara Marchetti, Raphaël Thyss, Giorgos Fitsialos, Mehdi Khaled, Lionel Larribère, Corine Bertolotto, Thierry Virolle, Pascal Barbry, Jacques Pouysségur, Gilles Ponzio and Robert Ballotti. Hypoxia Inducible Factor 1 alpha is a new target of Microphthalmia Associated Transcription Factor (MITF) in melanoma cells. J Cell Biol. 2005 Jul 4;170(1):49-59. Gaggioli C. Deckert M., Robert G., Abbe P., Batoz M., Ehrengruber M.U., Ortonne J-P, Ballotti R. & Tartare-Deckert, S. HGF induces. fibronectin matrix synthesis in melanoma cells through MAP kinase-dependent signaling pathway and induction of Egr-1. Oncogene 2005 Feb 17;24(8):1423-33. Marella M, Gaggioli C., Batoz M., Deckert M., Tartare-Deckert S. & Chabry J. Pathological prion protein exposure switches on neuronal MAP-kinase pathway resulting in microglia recruitment. J Biol Chem. 2005 Jan 14;280(2):1529-34. Cédric Gaggioli, Roser Buscà, Patricia Abbe, Jean-Paul Ortonne, and Robert Ballotti. Expression of Microphthalmia-associated Transcription Factor (MITF) is Required but does not Induce Melanogenesis in mammalian melanocytic cells. Pigment Cell Res. 2003 Aug;16(4):374-82.Presentation
Summary
Please note that about 30 % human cancers are basal and squamous cell carcinomas
Figure 2
In situ labellings are as indicated (Ki67, left; Cyclin D1 right) and show high rates of cell multiplication in epithelial invasions within the stroma.Research Project
Research Project
Research Team
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MENEGUZZI Guerrino, DR1 Inserm, Team Leader - Co-direction
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AL-GARAGHULI Sahar, PhD student, CNOUSTel : + 33 (0)4 93 37 77 47, Mail :
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ESTRACH Soline, CR1 InsermTel : + 33 (0)4 93 37 77 47, Mail :
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GACHE Yannick, CR1 InsermTel : + 33 (0)4 93 37 77 47, Mail :
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ROUANET Sophie, PhD StudentTel : + 33 (0)4 93 37 76 70, Mail :
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LACOUR Jean-Philippe, PU-PH CHU NiceTel : + 33 (0)4 92 03 64 89, Mail :
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PERRIN Chistophe, PH, CHU NiceTel : + 33 (0)4 93 37 XX XX, Mail :
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Patents
2003 : Thierry Magnaldo, A. Sarasin :« Use of the CD24 marker gene for the selection of keratinocyte stem cells transformed by exogenous sequences »International patent nbr WO 03004655, 2003.
Publications
Frechet, M., Warrick, E., Vioux, C., Benhamou, S., Spatz, A., Sarasin, A., Bernerd, F. and Magnaldo, T. (2008) Over expression of matrix metalloproteinase 1 in dermal fibroblasts from DNA repair deficient / cancer prone xeroderma pigmentosum group C patients, Oncogene, 27, 5223 - 5232.
Brellier, F., Valin, A., Chevallier-Lagente, O., Gorry, P., Avril, M-F., and Magnaldo, T. Ultraviolet responses of Gorlin syndrome primary skin cells. British J. Dermatol. 2008, 59, 445-452.
Bergoglio, V., Emilie Warrick, E., Chevallier-Lagente, O., and Magnaldo, T. (2008) Cutaneous gene therapy : the graft takes / Thérapie génique cutanée : la greffe prend / Médecine/Science, 24, 607-14.
Bergoglio, V., Larcher, F., Chevallier-Lagente, O., Bernheim, A., Danos, O., Sarasin, A, Del Rio, M.D., and Magnaldo, T. 2008 Safe Selection of Genetically Manipulated Human Primary Keratinocytes with Very High Growth Potential Using CD24. Mol. Therapy. 12, 2186-2193.
Groisman, R., Kuraoka, I.,Chevallier, O., Gaye, N., Magnaldo,T. , Tanaka, K.,5, Harel-Bellan, A., Kisselev, AF., and Yoshihiro Nakatani (2006) CSA-dependent degradation of CSB by the ubiquitin-proteasome pathway establishes a link between complementation factors of the Cockayne syndrome. Genes and Dev., 20 , 1429-1434.
Brellier, F., Marionnet, C., Chevallier-Lagente, O., Toftgard, R., Mauviel, A., Sarasin, A. and Magnaldo, T. (2004). Ultraviolet radiation represses PATCHED transcription in human epidermal keratinocytes through an AP1-depedent process. Cancer Res 64, 2699-2704. Baby Team Cédric Gaggioli
Presentation
In primary tumors, cancer cells are immersed in a complex microenvironment network comprising numerous cells, including carcinoma associated fibroblasts (CAF). Tumor-stroma crosstalk plays a major role in carcinoma development and invasion. We have previously demonstrated the crucial role of Janus Kinase dependent signaling pathway, in CAF cells, for matrix remodeling and tracks formation for cancer cells to invade. However, how “normal” resident fibroblasts are converted into pro-invasive CAF cells remains an open question. Malignant cells themselves promotes carcinogenesis through cytokines and growth factors secretion that influence stroma cells behaviour, which in return influence carcinoma cells invasion and malignancy. In line with our previous research, we aim at uncovered signaling pathways and molecules that govern stromal fibroblasts and carcinoma cells crosstalk during invasion, which could lead to the identification of therapeutic target for aggressive and metastatic cancer.
Research Project
The project presents two major axes focused on :
Investigation of the signaling pathways that in SCC cells mediates the pro-invasive conversion of resident fibroblasts
We aim at identifying pathway in pro-invasive CAFs conversion from primary human dermal fibroblasts (HPF) isolated from children foreskin. Using in vitro matrix remodeling and three-dimensional organotypic invasion assay.
ALBRENGUES Jean, PhD StudentTel : + 33 (0)4 93 37 77 53, Mail :
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BOURGET Isabelle, IE1 InsermTel : + 33 (0)4 93 37 77 53, Mail :
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Team Thierry MAGNALDO & Guerrino MENEGUZZI & Baby Team Cédric GAGGIOLI
