Chemical tools to investigate the role of sphingolipids in disease

226 Pages·2016·27.65 MB·English

DOCTORAL PROGRAMME IN ORGANIC CHEMISTRY

Chemical tools to investigate the
role of sphingolipids in disease

Yadira Ordóñez Vivanco

Ph.D. Candidate

Gemma Fabriàs Domingo Gemma Triola Guillem

Thesis Supervisor Thesis Supervisor

Department of Biomedicinal Department of Biomedicinal
Chemistry (iQAC-CSIC) Chemistry (iQAC-CSIC)

Anna Grandas Sagarra

Tutor

Department of Inorganic and
Organic Chemistry
University of Barcelona

This work was supported by grants from the Spanish Ministries of Science
and Technology (SAF2011-22444) and Economy and Competitiveness
(CTQ2014-54743-R) and a fellowship from the Secretaría de Educación
Superior, Ciencia, Tecnología e Innovación del Ecuador (SENESCYT).

ACKNOWLEDGMENTS
I would like to express my sincere gratitude and appreciation to my thesis
advisor, Dr. Gemma Fabriàs and Dr. Gemma Triola, for their supervision
and support throughout my research work, in whom from the first day I
found patience, dedication and motivation.

I would also like to thank Dr. Josefina Casas, Dr. Antonio Delgado and Dr.
José Luis Abad for their advice and suggestions during the development of
this thesis.

I am grateful to Dr. Jordi García, Dr. Angel Montaña, Dr. Pilar Marco for
acting as members of my committee and for reviewing this thesis. I am also
grateful to my thesis tutor Dr. Anna Grandas.

I also thank my friends and all the people I had the pleasure to share great
moments during my life as Ph.D student. My lab partners, with whom I
have shared every working day, every joy and disappointment, which are
the common factor of all experimental work.

Finally, I would like to thank and dedicate this work to the people who have
accompanied and supported me unconditionally in all stages of my life; my
parents from which I got example of strength and perseverance; my sister
and my best friend, for always believing in me, I thank her for being with
me those long hours from the distance.

To all, thank you very much!

CONTENT

INTRODUCTION

1. Sphingolipids 3
1.1. Sphingolipid metabolism in mammals 4
1.2. Compartmentalization and transport of Sphingolipids 6
1.3. Sphingolipids in disease 8
1.4. Sphingolipids in autophagy 12
2. Dihydroceramides and dihydroceramide desaturase 16
2.1. Des1 inhibitors 18
2.1.1. γ-tocopherol and γ-tocotrienol 19
2.1.2. XM462 20
3. Ceramidases 21
3.1. Acid ceramidase 22
3.2. Acid ceramidase and human diseases 22
3.2.1. Farber disease (FD) 22
3.2.2. Acid Ceramidase and cancer 23
3.2.3. Acid Ceramidase and other complex disease 25
3.2.4. Acid ceramidase inhibitors 25
3.2.4.1. Ceramide-based inhibitors 25
3.2.4.2. Inhibitors structurally unrelated to ceramide 28
3.2.4.3. Chaperones 29
4. Activity based probes 30
5. Click Chemistry in chemical biology of sphigolipids 32

OBJECTIVES 37

RESULTS

Chapter I
Validation of the role of dihydroceramides as mediators of
41
autophagy in cancer cell models using metabolic precursors
Chapter II
Validation of the role of dihydroceramides as mediators of
59
autophagy in cancer cell models using Des1 inhibitors
CHAPTER III
Development of an activity based probe for acid ceramidase 103
detection in cells

GENERAL DISCUSSION 129

CONCLUSIONS 147

REFERENCES 153

RESUMEN 173

APPENDIX I 199