Role and prognostic importance of regulatory T cells in lung cancer patients, according to the ...

276 Pages·2015·6.31 MB·French

by  Higgs, Brandon

Université Pierre et Marie Curie
Ecole doctorale ED394 : Physiologie et Physiopathologie et
thérapeutique
UMRS1138
Thèse de doctorat d’immunologie

Présentée par :
Priyanka DEVI
Pour obtenir le titre de Docteur de l’université Pierre et Marie Curie
Role and prognostic importance of regulatory T cells in lung cancer
patients according to the presence of tertiary lymphoid structures
Présentée et soutenue publiquement le 5 Octobre 2015

Devant un jury composé de :
Prof. François Lemoine : Président du Jury
Prof. Eric Tartour : Rapporteur
Dr. Bruno Lucas : Rapporteur
Dr. Christine Caux : Examinatrice
Prof. François Ghiringhelli : Examinateur
Prof. Catherine Sautés-Fridman : Invitée
Dr. Marie-Caroline Dieu-Nosjean : Directrice de thèse
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Acknowledgement

First, I would like to thank Prof. Herve Fridman and Current director Mr.
Pascal Ferre, for accepting me to the Centre de recherche de Cordeliers.
I would like to thank you, Prof. Herve Fridman, for his scientific advices and
valuable discussions during the lab-meetings which helped me to get insights of tumor
immunology and helped this work to improve qualitatively.
I thank Prof. Catherine Sautes-Fridman for giving me an opportunity to join
this laboratory as PhD student through the Erasmus Mundus scholarship program. I
thank her for the kindness and scientific discussions during the lab meetings. I thank her
for giving me an opportunity to talk in the departmental day. Since, my first day in the
lab, I am so pleased with your and Herve’s love about Indian food and culture. I thank
their affection about India and I hope they will continue loving India.
I thank Prof. François Lemoine for accepting to be president of the jury. I
thank Prof. Eric Tartour and Dr. Bruno Lucas for reviewing the thesis manuscript. I thank
Dr. Christine Caux and Prof. François Ghiringhelli for accepting to be examiners of the
thesis.
I would like to sincerely acknowledge Dr. Marie-Caroline Dieu-Nosjean, who
accompanied me throughout this journey. I thank her, for her patience and the
confidence she showed in me. Her scientific parenthood helped me to understand the
tumor immunology, which was quite a foreign field for me on my arrival in lab. I thank
her for giving me an opportunity to learn so many new techniques and helping me to be
an independent in performing the experiments and understanding the results. I cannot
forget the long meetings with the designing experiments or discussing the results
(especially writings on the white board!). This thesis could not have been better without
her motivation and immense knowledge which widen my research from various
aspects.
My sincere thanks also go to Dr. Jean-Luc Teillaud and Prof. Isabelle Cremer,
who integrated me in this laboratory and allowed me to access the laboratory and
research facilities. I also thank them for the kind follow up my work during these four
years and their scientific contributions which helped this work to improve. I especially
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thank Jean-Luc for sharing not only scientific interests, but also the political, geographic,
historic and artistic views with me. I would like to thank Dr. Sophie Siberil for her
contributions in the lab meeting and her kindness.
I thank Dr. Audrey Lupo, Dr. Diane Damotte, Dr. Marco Alifano and the team
of clinicians and pathologists at Hotel Dieu and Cochin Hospital; also to the Dr. Pierre
Validire and the team at the Institute Mutualiste Montsouris hospital for their valuable
help for the surgical specimen and the clinic-pathological datas of the cancer patients.
Without their help this project could not have been moved ahead.
I would like to thank CICC team for their grand technical support. My thanks
goes to Hélène and Estelle for their great help for the flow cytometry and I thank
Christophe for his help in the imaging. I wish Estelle a very good luck for end of her
thesis.
I thank my dear friends (“100% women’s team Marie Caroline”) i.e. Claire,
Samantha, Helene, Claudia and Myriam for their love and support throughout this
thesis. I would like to thank a lot of my dear post-doc Claire, for her incredible help
during the experiments and her kindness. Thanks for sharing the Calopix pain with me
in all these years! Your hard work and perfectionism have always been inspiring for me!
I would like thank Sam for her kindness and motherhood during the molecular biology
experiments. Thank you for motivating me with your kind words “ca vas aller” during
stressful times! I would like to thank my dear friend Helene for her happy and
humorous nature. Thank you for making me laughs all the time! I wish you a very good
luck for your thesis. I would like to thank you Claudia, my English speaking companion
in the team for your help and support. Thank you for teaching me the Ingenuity
software and it was nice to discuss with you a lot about gene expression data. Gracias
mi amiga peruana! I would like to thank Myriam for her help in the last few months of
my thesis and I hope you will continue liking the Indian food. I would like to thank my
former DC-Lamp companion, “Mr. DOC GOC” for his help and support during the initial
phase of my thesis. I hope he will be successful in his adventures in New-York.
I would also like to thank other post-docs Sarah, Jerome, Pauline in the lab.
Your presence in the lab was helpful for me! I thank you Sarah for introducing the
nanostring technology and most importantly for giving pleasure to hear some American
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English in the lab! I thank you Jerome for always helping me with antibodies and also
thank you for your curiosity with the Tregs project. I thank you Pauline for your
kindness and help. I wish a good luck to new postdocs Mihaela and Angella for their
projects in the lab.
I would like to thank the IHC “super girls” Ben Ben, Tick Tick, Laetitia, Hanane,
Estephania and Marion, for their “super energy” to organize parties and fun times. I
thank them for keeping the IHC lab always in a full energized mode. I thank you a tennis
girl Tessa for your kindness and your help in the molecular biology experiments. Thanks
for sharing and teaching me how to break correctly the “Noix”! I hope you will visit the
Himalayas (Mt. Everst) and of course me:P one day! I also thank the Lucie, Nathalie
Josseaume, Tania, Kris and Melanie for their help. I would like to thank Gabriela and
Bernhard for their advice during the bioinformatics data analyses.
I would like to thank my dear Indian friend Saradiya for accompanying me in
the initial years of the thesis. I was always lucky, to have you before and later Shambhu,
to share Indian food and to speak in Hindi while learning French! I wish you very good
luck Shambhu, to end of your thesis! Also, I would like to thank other PhD students in
the lab who were sailing in the same boat with me! I would like to thank Nicolas,
Etienne, Nicolas Merle, Claire Deligne, Mikael, and Benoit for their friendship
throughout my thesis.
I would like to thank Dr.Lubka Roumenina, Dr. Véronique Fremeaux-Bacchi
and Marie-Agnes Dragon-Durey for their scientific discussions in lab meetings.
I would like to thank the past and present M2 students in lab for keeping the
lab young and fresh! Thanks to the past members of the lab Jeremy Cosette, Romain
Remark, Caroline, Claire Galand and Duy for their help.
Je vous remercie de Nathalie et Eliane pour être mère à nous tous! La
manière vous soignez pour moi a toujours été incroyable! Je remercie Johanna et Lamia
pour toute l'aide administrative que vous avez fait pendant toutes ces années! Je
remercie Jasmina pour amener l'humour pendant le déjeuner et en général aussi! Sans
vous tout le laboratoire ne peut pas fonctionner sans problèmes !
I would like to thank our collaborators in this project Sadrine Katsahian and
Sylvain Leveugle from team 22 CRC for their help in the statistical analyses. I thank
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David Gentien and Benoit Albaud for their hospitality at the Curie institute. I also thank
Emmanuel Donadieu and Houcine Bougherara for their help and hospitality in Cochin
institute.
I would like to thank my professor Dr. Kanchanganga Gandhe for her
enormous support and love during all this phase of my work. Her kind words have
always motivated me all these years. I thank my dearest friend Rhucha for her
incredible love and support. I would like to thank my friends in India Tejashree,
Revatee, Sharvari, Kalyani, Mihir for their priceless love, friendship and their continuous
support, although we were far from each other in all these thesis years.
I would like to thank my parents, parent’s in-laws and my younger brother for
their precious love, their kindness and their continuous support throughout this phase
of my life. Words feel shorter to thank them!
Last but not least, I thank deep from my heart to love of my life, my husband,
Swanand who made this journey pleasant for me. I was lucky to have you with me in
this beautiful and romantic city, Paris. There comes a time in PhD when you have to
constantly remind yourself that not everyone is running the same race, and that you're
only competing with who you were yesterday. In all those times, I thank you for being
with me, for showering your relentless love and tremendous support.

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“Life is not easy for any of us.
But what of that? We must have
perseverance and above all
confidence in ourselves. We
must believe that we are gifted

for something and that this

thing, at whatever cost, must be

attained”

-Marie Curie

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To my parents,
For their relentless love for me…

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Table of contents
Acknowledgement ................................................................................................... 2
Index of figures ...................................................................................................... 11
Index of tables ........................................................................................................ 12
Abstract 13
Abbreviations ......................................................................................................... 14
A. Introduction .................................................................................................... 17
1. Immune system and tumors: a complex discourse ....................................... 17
1.1. Origin of the concept of tumor microenvironment....................................... 17
1.2. Origin and concept of immune surveillance ................................................. 19
1.2.1. Immunoediting: 3 `E’ concept ...................................................................... 20
1.3. Tumor microenvironment: a complex interactome ...................................... 22
1.3.1. Characteristics of contexture ........................................................................ 24
1.3.2. Cancer associated TLS ................................................................................. 25
1.3.2.1. General characteristics of TLS ........................................................ 25
1.3.2.2. Formation of TLS ............................................................................ 26
1.3.3. TLS in anti-tumor immune response ............................................................ 28
1.4. Infiltration of immune cells in solid tumor: a strong prognostic marker ..... 31
1.4.1. Prognostic importance of the TLS in cancer ...................................... 32
2. Tregs: Key Regulators of anti-tumor immune response............................... 35
2.1. Discovery and features of regulatory T cells ................................................ 35
2.2. Regulatory T cell subsets.............................................................................. 38
2.3. Regulatory mechanisms exerted by Tregs .................................................... 41
2.3.1. Inhibitory cytokines ...................................................................................... 42
2.3.2. Suppression by cytolysis .............................................................................. 43
2.3.3. Suppression by metabolic disruption............................................................ 44
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2.3.4. Cell to cell contact dependent suppression: Involvement of co-stimulatory and
co-inhibitory signals............................................................................................... 45
2.4. How many mechanisms do Tregs need? Treg plasticity .............................. 51
2.5. Infiltration, differentiation and activation of Tregs in tumor microenvironment
53
2.5.1. Infiltration of Tregs in tumor microenvironment ......................................... 53
2.5.2. Expansion and activation of Tregs ............................................................... 55
2.5.3. Antigen specificity of Tregs in cancer .......................................................... 55
2.6. Tregs in cancer: ambiguity in prognostic importance .................................. 56
3. Tregs and immunotherapy: a blessing in disguise? ...................................... 60
4. Lung cancer: a study model .......................................................................... 66
4.1. Etiology and histology of the lung cancer .................................................... 67
4.2. TNM classification and survival of patients ................................................. 68
4.3. Treatment of lung cancer patients ................................................................ 71
4.4. Era of combined therapies: promising for NSCLC ...................................... 72
B. Hypothesis and objectives .............................................................................. 75
C. Results 80
References 106
Tregs in advanced stage chemotherapy treated lung cancer patients .................. 141
D. Discussion ..................................................................................................... 146
TLS in lung tumors: centers of the protective immune responses ....................... 146
Infiltration, activation of Tregs in cancer microenvironment .............................. 147
Anti-inflammatory role of Tregs in lung cancer .................................................. 150
Tregs in TLS: Proponents or opponents?............................................................. 152
Expansion of the specific subsets of Tregs in TLS .............................................. 156
Modulation of the Treg phenotype in the neoadjuvant chemotherapy treated lung
cancer ................................................................................................................... 157
Prognostic outcome of Tregs in lung cancer patients .......................................... 158
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E. Limitations of this study ............................................................................... 161
F. Conclusion and perspectives ........................................................................ 162
G. References .................................................................................................... 165
Publication bibliography ...................................................................................... 165
H. Annex 199

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