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Prostate Cancer The supramolecular organization of cancer metabolism: From macromolecular crowding to metabolic reprogramming underlying cancer metastasis and drug resistance Cristina Balcells Nadal ADVERTIMENT. La consulta d’aquesta tesi queda condicionada a l’acceptació de les següents condicions d'ús: La difusió d’aquesta tesi per mitjà del servei TDX (www.tdx.cat) i a través del Dipòsit Digital de la UB (diposit.ub.edu) ha estat autoritzada pels titulars dels drets de propietat intelꞏlectual únicament per a usos privats emmarcats en activitats d’investigació i docència. No s’autoritza la seva reproducció amb finalitats de lucre ni la seva difusió i posada a disposició des d’un lloc aliè al servei TDX ni al Dipòsit Digital de la UB. No s’autoritza la presentació del seu contingut en una finestra o marc aliè a TDX o al Dipòsit Digital de la UB (framing). Aquesta reserva de drets afecta tant al resum de presentació de la tesi com als seus continguts. En la utilització o cita de parts de la tesi és obligat indicar el nom de la persona autora. ADVERTENCIA. La consulta de esta tesis queda condicionada a la aceptación de las siguientes condiciones de uso: La difusión de esta tesis por medio del servicio TDR (www.tdx.cat) y a través del Repositorio Digital de la UB (diposit.ub.edu) ha sido autorizada por los titulares de los derechos de propiedad intelectual únicamente para usos privados enmarcados en actividades de investigación y docencia. No se autoriza su reproducción con finalidades de lucro ni su difusión y puesta a disposición desde un sitio ajeno al servicio TDR o al Repositorio Digital de la UB. No se autoriza la presentación de su contenido en una ventana o marco ajeno a TDR o al Repositorio Digital de la UB (framing). Esta reserva de derechos afecta tanto al resumen de presentación de la tesis como a sus contenidos. En la utilización o cita de partes de la tesis es obligado indicar el nombre de la persona autora. WARNING. On having consulted this thesis you’re accepting the following use conditions: Spreading this thesis by the TDX (www.tdx.cat) service and by the UB Digital Repository (diposit.ub.edu) has been authorized by the titular of the intellectual property rights only for private uses placed in investigation and teaching activities. Reproduction with lucrative aims is not authorized nor its spreading and availability from a site foreign to the TDX service or to the UB Digital Repository. Introducing its content in a window or frame foreign to the TDX service or to the UB Digital Repository is not authorized (framing). Those rights affect to the presentation summary of the thesis as well as to its contents. In the using or citation of parts of the thesis it’s obliged to indicate the name of the author. UNIVERSITAT DE BARCELONA FACULTAT DE FARMÀCIA I CIÈNCIES DE L’ALIMENTACIÓ PROGRAMA DE DOCTORAT EN BIOTECNOLOGIA The supramolecular organization of cancer metabolism: From macromolecular crowding to metabolic reprogramming underlying cancer metastasis and drug resistance Cristina Balcells Nadal 2019 Facultat de Farmàcia i Ciències de l’Alimentació PROGRAMA DE DOCTORAT EN BIOTECNOLOGIA Departament de Ciència de Materials i Química Física Departament de Bioquímica i Biomedicina Molecular The supramolecular organization of cancer metabolism: From macromolecular crowding to metabolic reprogramming underlying cancer metastasis and drug resistance Memòria presentada per Cristina Balcells Nadal per optar al títol de Doctora per la Universitat de Barcelona Marta Cascante Serratosa Francesc Mas i Pujadas Codirectora Codirector Josefa Badia Palacín Cristina Balcells Nadal Tutora Doctoranda Index Index 1. Introduction ...................................................................................................... 3 1.2.1. Cancer and cell cycle .................................................................................. 5 1.2.2. Cancer and apoptosis ................................................................................. 7 1.3.1. Overview of cancer metabolism ............................................................... 10 1.3.2. Regulation of cancer metabolism ............................................................. 15 1.3.2.1. Pro-anabolic signaling networks ....................................................... 15 1.3.2.2. Pro-catabolic signaling networks ...................................................... 16 1.3.2.3. Regulation of redox balance ............................................................. 17 1.3.2.4. Fine-tuning other aspects of cell metabolism ................................... 18 1.3.3. Glycolysis and the Warburg effect ........................................................... 19 1.3.4. Tricarboxylic acid cycle and oxidative phosphorylation ........................... 21 1.3.5. Amino acid metabolism ............................................................................ 23 1.3.6. Glutamine metabolism ............................................................................. 24 1.3.7. Redox balance and ROS metabolism ........................................................ 26 1.3.8. Pentose phosphate pathway .................................................................... 27 1.3.9. One-carbon metabolism ........................................................................... 28 1.3.10. Urea cycle .............................................................................................. 29 1.3.11. Polyamine metabolism ......................................................................... 30 1.3.12. Lipid metabolism ................................................................................... 33 1.4.1. Metastasis and epithelial to mesenchymal transitions ............................ 35 1.4.2. Tumor initiation, stemness, and plasticity ............................................... 38 1.4.3. Interplay between the EMT and CSC programs ....................................... 39 1.4.4. Metabolic reprogramming associated to metastasis and invasion ......... 40 1.4.5. Metabolic reprogramming associated with stemness ............................. 45 1.5.1. Mechanisms of drug resistance ................................................................ 46 1.5.2. Metabolic reprogramming associated with resistance ............................ 51 1.5.3. Platinum-based chemotherapy ................................................................ 52 1.5.4. Platinum (IV) compounds ......................................................................... 53 1.5.5. Mechanisms of platinum resistance ......................................................... 54 1.6.1. Excluded volume effects ........................................................................... 55 1.6.2. Macromolecular crowding, metabolism, and cancer ............................... 57 1.7.1. Histological classification and subsets of prostate cancer ....................... 59 1.7.2. Oncogenic signaling and treatment of prostate cancer ........................... 61 1.7.3. Prostate cancer metabolism ..................................................................... 62 1.7.4. Prostate cancer cell models...................................................................... 64 1.8.1. Oncogenic signaling and treatment of colorectal cancer ........................ 66 1.9.1. Metabolomics ........................................................................................... 67 1.9.2. Metabolic target predictions .................................................................... 68 2. Objectives ....................................................................................................... 73 3. Materials and methods .................................................................................... 77 3.1.1. Prostate cancer cell lines .......................................................................... 77 3.1.2. Colorectal cancer cell lines ....................................................................... 78 3.1.3. Other cell lines .......................................................................................... 78 3.2.1. Direct cell counting ................................................................................... 79 3.2.2. MTT and Hoechst assays .......................................................................... 79 3.7.1. Determination of metabolites in cell pellets ............................................ 84 3.7.2. Determination of metabolites in culture media ...................................... 85 3.8.1. Lactate dehydrogenase ............................................................................ 86 3.8.2. Glutamate dehydrogenase ....................................................................... 86 4. Results and discussion ..................................................................................... 95 4.1.1. Introduction .............................................................................................. 95 4.1.2. Results and discussion .............................................................................. 97 4.1.2.1. Proliferation, EMT, and stem cell traits are uncoupled in CRPC ....... 97 4.1.2.2. Metastatic prostate CSCs display enhanced glutamine avidity and their proliferation and tumorigenic capacity can be suppressed through glutamine deprivation ....................................................................................... 102 4.1.2.3. CRPC e-CSC phenotypes present enhanced methionine, lysine and total amino acid consumption rates.................................................................
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