Institut de Génétique et de Biologie Moléculaire et Cellulaire

Daniel Metzger

Description

Niveau: Supérieur, Doctorat, Bac+8
Institut de Génétique et de Biologie Moléculaire et Cellulaire UMR CNRS 7104/INSERM U596/ULP/Collège de France Thèse présentée à l'URF des Sciences de la Vie et de la Santé de l'Université Louis PASTEUR - Strasbourg I. Discipline Biologie Moléculaire pour obtenir le titre de Docteur de l'Université Louis PASTEUR de Strasbourg par Eduardo Castañeda Saucedo Role of the Retinoid X Receptor ? (RXR?) and its Heterodimeric Partners during Skin Carcinogenesis Directeur: Daniel Metzger Co-directeur: Pierre Chambon Date de soutenance 1 Juillet 2004 Jury: Prof. Pierre Chambon Dr. Daniel Metzger Prof. Johan Auwerx Prof. Peter Angel Dr. Lluis Fajas

• factor histone


• spindle cell


• ppar? during


• vitamin d3 vitamin


• carcinoma basement


• nuclear receptors


• receptor focal


• skin carcinogenesis


• igbmc

Sujets

• Metzger
• Louis Pasteur University
• Lourdes
• Chambon
• Institut national de la santé et de la recherche médicale
• La Ligue nationale contre le cancer

Informations

• Publié par: profil-zyak-2012
• Publié le: 01 juillet 2004
• Nombre de lectures: 112
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Institut de Génétique et de Biologie Moléculaire et Cellulaire
UMR CNRS 7104/INSERM U596/ULP/Collège de France
Thèse
présentée à
l'URF des Sciences de la Vie et de la Santé
de l'Université Louis PASTEUR - Strasbourg I.
Discipline Biologie Moléculaire
pour obtenir le titre de
Docteur de l'Université Louis PASTEUR de Strasbourg
par
Eduardo Castañeda Saucedo
Role of the Retinoid X Receptor α (RXRα) and its Heterodimeric
Partners during Skin Carcinogenesis
Directeur: Daniel Metzger
Co-directeur: Pierre Chambon
Date de soutenance 1 Juillet 2004
Jury: Prof. Pierre Chambon
Dr. Daniel Metzger
Prof. Johan Auwerx
Prof. Peter Angel
Dr. Lluis FajasAcknowledgments
To Prof. Pierre Chambon and Dr. Daniel Metzger for giving me the
opportunity to prepare my PhD. thesis in their laboratory, for their
scientific and personal support, for their critical comments and for all
what I have learned from them.
I would like to thank Prof. Johan Auwerx, Prof. Peter Angel and Dr.
Lluis Fajas for accepting to be part of my Ph.D. jury.
To Dr. Patricio Gariglio, for all his support before and during my Ph.D.,
for his critical comments, and most important for his friendship.
To Arup Indra, my colleague, collaborator and friend, with whom I have
worked during my Ph.D., for his advice, critical discussions and for all
the good time we spent together.
To Mei Li, for her scientific support, suggestions and critical comments,
and in particular for her nice friendship.
Special thanks to Monique Duval, for all her inconditional support with
animal work.
To all the current and former members of the laboratory, Jean-Marc,
Takeshi, Meng, Ming, Philipp, Reiko, Yasunari, Nathalie, Celine,
Isabelle, Romain, Pierre, Michael, Michal, Ali, Bernadette, Christelle,
Jean-François, Monica, Martha, Wojtek, Emilie, Valèrie, (and all those
whom I may forget) for their critical comments, their support, and their
friendship.
To all people from the IGBMC common facilities (histology service,
oligonucleotide synthesis service, animal facility, photographic service,
informatics service). Special thanks to Kristina, Nadia and Bernard, for
their technical support. To all other people who make our work possible
at the IGBMC.
Special thanks to Collete Kutschis, for all her support and friendship.
To all my friends and colleagues, current and former members of the
IGBMC, thanks to whom my time at the IGBMC has been so pleasent.
Special thanks to the "Ligue Contre le Cancer, comité du Bas-Rhin"
for the financial support during my thesis.Finally, thanks to Lourdes, my wife and my friend, for being
there all the time, in the god and the not so good times, for her
motivation, her support and her love. Thank you.Abreviations
AF-1 activation function 1
AF-2 activation function 2
AK actinic keratosis
AP-1 activator protein 1
BCC basal cell carcinoma
BM basement membrane
CE cornified envelope
DBD DNA binding domain
DMBA 7-12-dimethylben(a)anthracene
ECM extra cellular matrix
ER estrogen receptor
FC focal carcinoma
GM-CSF granulocyte macrophage colony stimulator factor
HAT histone acetyl transferase
HDAC histone deacetylase.
HMT histone methyl transferase
IHC immunohistochemistry
IL-1 interleukin 1
IRS inner root sheath
ISC in situ carcinoma
ISH in situ hybridisation
KGF keratinocyte growth factor
K13 keratin 13
K14 keratin 14
LBD ligand binding domain
LXR liver X receptor
MG's melanocytic growths
NSAID non steroidal anti-inflammatory drugs
NR nuclear receptor
ORS outer root sheath
PMD post mitotic differentiating
PPAR peroxisome proliferator activated receptor
RA retinoic acid
RAR retinoic acid receptor
RT-PCR reverse transription coupled polymerase chain reaction
rtTA reverse tetracycline-controlled transactivator
RXR retinoic X receptor
SCC squamous cell carcinoma
SCF stem cell factor
Sp-1 specificity protein-1
SpCC spindle cell carcinoma
TA transit amplifying
Tam tamoxifen
Tet tetracyclin
TPA 12-O-tetradecanoyl-phorbol-13-acetate
UV ultraviolet
VD vitamin D33
VDR vitamin D receptorTable list.
Table 1. Common Neoplasm of the Skin .................................................................................. 11
Table 2. Mouse models of skin cancer ..................................................................................... 14
Table 3. Synthesis of the classical lipophilic hormones........................................................... 23
Table 4. Human and Mouse nuclear receptors........................................................................ 26
Table 5. Expression of NRs in the skin..................................................................................... 32
ep/-Table 6. Summary of skin carcinogenesis in Tg.AC/RXRα mice......................................... 61
Table 7. Summary of RXRα and its heterodimeric partners functions during skin carcinogenesis..... 70
Figure list.
Figure 1. Structure of human skin.............................................................................................. 2
Figure 2. Keratinocyte proliferation unit................................................................................... 7
Figure 3. The chemical skin carcinogenesis model.. ............................................................... 15
Figure 4. The Tg.AC model...................................................................................................... 17
Figure 5. The Cre-loxP system of homologous recombination................................................ 19
Figure 6. Strategy to generate a tamoxifen-induceble Cre recombinase. ............................... 20
T2Figure 7. Model of the mechanism of Cre-ER -mediated recombination in keratinocytes.... 21
Figure 8. Structure of the major lipophylic hormones...................................................................... 24
Figure 9. Modular structure of the NRs................................................................................... 25
Figure 10. Nuclear receptors response elements..................................................................... 27
Figure 11. Mechanism of NR transcription regulation............................................................ 29
ep-/-Figure 12. Phenotype of RXRα mice................................................................................... 35
Figure 13. Model for integration of nuclear receptors action in the skin. ............................. 39
Figure 14. Role of PPARα during skin carcinogenesis 46
Figure 15. Selective ablation of RXRα in epidermal keratinocytes of adult Tg.AC ................ 53
Figure 16. Selective ablation of RXRαtinocytes of Tg.AC mice during skin
morphogenesis.......................................................................................................................... 56
ep-/-(c)Figure 17. Formation of skin papillomas in RXRα mice in response to a single DMBA
application................................................................................................................................ 59
ep-/-(c)Figure 18. Formation of melanocytic growths (MGs) in RXRα mice in response to a
single DMBA application......................................................................................................... 60
ep-/-(c)Figure 19. Phenotypic changes in the skin of Tg.AC/RXRα mice............................................. 63
ep-/-Figure 20. Skin carcinogenesis in PPARγ mice......................................................................... 74Résumé
La peau des Mammifères comporte, sur sa face externe, l'épiderme, tissu épithélial
pavimenteux (squameux) stratifié et kératinisé (corné), composé de kératinocytes (espèce
majoritaire), de cellules pigmentaires (mélanocytes), de cellules immunitaires (cellules de
Langerhans) et de mécanorécepteurs (cellules de Merkel) (Kanitakis, 2002). L'épiderme est
ancré, via une lame basale, à du tissu conjonctif dense fibroélastique, le derme (Watt, 2002).
Ce tissu largement vascularisé, composé de fibroblastes et de cellules immunitaires
(mastocytes et histiocytes), repose sur l'hypoderme, association de tissu conjonctif lâche et de
tissu adipeux (Kanitakis, 2002). Des invaginations épidermiques dans les tissus sous-jacents
constituent des crêtes épidermiques ou forment les annexes épidermiques telles que les poils,
les glandes sébacées (holocrines), ou les glandes sudoriférantes (apocrines) (Fuchs, 1998;
Larsen, 1993; Mackenzie, 1975). La peau joue des fonctions très diverses, elle a des fonctions
métaboliques, sensorielles, thermorégulatrices, immunitaires, et elle représente une barrière
contre la déshydratation et les agressions externes (radiations UV, friction, parasites . . . )
Les rétinoïdes, métabolites actifs de la vitamine A, ont des effets pléïotropes sur l'homéostasie
et le développement des Vertébrés. La peau est un des tissus cibles des retinoïdes. En effet,
une carence ou un excès systémique en vitamine A