2017-03-16 Medical sciences Medicine Medizin Intermediärstoffwechsel opus:7223 OCR German https://doi.org/10.17192/z2017.0200 Publikationsserver der Universitätsbibliothek Marburg Universitätsbibliothek Marburg Philipps-Universität Marburg 194 application/pdf ths Prof Dr. Müller Rolf Müller, Rolf (Prof Dr.) 2017 Identification of metabolic pathways in human ovarian cancer cells with potential therapeutic relevance A characteristic feature of tumor cells is their altered intermediary metabolism. Tumor cells frequently exhibit increased glucose uptake and lactate production even in the presence of oxygen (aerobic glycolysis, "Warburg effect"). Other metabolic pathways activated in tumor cells, including the degradation of glutamin as an anaplerotic reaction of the citric acid cycle ("glumaminolysis"), de novo fatty acid synthesis and fatty acid degradation via β-oxidation also have attracted particular attention. These altered metabolic pathways represent promising potential pharmacologic targets for cancer therapy . Ovarian cancer is the most lethal of all gynaecologic malignancies with the development of ascites in the peritoneal cavity as a characteristic feature. The isolation of primary cancer cells from ascites provides a unique opportunity to investigate metabolic aberrations in this tumor entity. The aim of this study was the identification of tumor-specific metabolic pathways as potential targets for new therapeutic approaches. For this purpose, patient-derived ovarian carcinoma cells were analyzed in comparison to a number of established tumor cell lines and the non-transformed breast epithelial cell line MCF-10A. A prerequisite for this study was the characterization of the metabolic phenotypes by expression analysis and "metabolic profiling" using the Seahorse XFe analyzer. Towards this goal, the metabolic parameters OCR (oxygen consumption rate) and ECAR (extracellular acidification rate) as well as the corresponding metabolic capacities were measured under normal conditions and metabolic pressure, mainly induced by metabolic inhibitors. This work revealed a specific metabolic feature of the ovarian carcinoma cell line SKOV-3. ATP production from glucose by oxidative phosphorylation (OXPHOS) is blocked in these cells. Initially, this was hypothesized to be caused by a defect in TCA-cycle. However, this hypothesis is not compatible with the finding that SKOV-3 cells are able to sustain ATP recovery via OXPHOS over long periods of time. Therefore, a "fatty acid cycle" (FAC) was postulated as an alternative metabolic pathway. This pathway consists of the cyclic synthesis and degradation of fatty acids. The advantages of the FAC could be independence of external supply of substrates and increased anabolic capacity due to a shift of glucose metabolism towards pentose phosphate pathway. Due to the advantages of the FAC for proliferating cells, it is likely that this cycle is also operative in a variety of other cells. The reason for the abnormal SKOV-3 metabolism was in the end localised as a defect in pyruvate transport into the mitochondrial matrix, which is due to a reduced expression of Mitochondrial Pyruvate Carrier 1 gene (MPC1). Another important result was the finding of increased aerobic glycolysis and β-oxidation in all patient-derived ovarian carcinoma cells. By using inhibitors of tumor relevant metabolic pathways, the potential to interfere with the proliferation or survival of these cells was examined using the xCELLigence RTCA. It was shown that single inhibitors were able to reduce proliferation (probably unspecifically) only at excessive concentrations, but that the combination of different metabolic modulators at concentrations ineffective on their own resulted in significant synergistic effects. Especially the combination of Oxamate (lactate dehydrogenase inhibitor) with the inhibitors DCA (inhibitor of pyruvate dehydrogenase kinases; PDKs), AOA (inhibitor of aspartate transaminase and thus of the malate aspartate shuttle) and SB204990 (inhibitor of ATP citrate lyase) caused a distinct proliferation inhibition, including cell death. Nevertheless ovarian carcinoma cells from different patients differ in part substantially with respect to their metabolism and the anti-proliferative effects of inhibitors. Examples are differences in (i) the utilization of glutamine and pyruvate as the sole substrate, (ii) the dependence of PDKs, lactate dehydrogenase and aspartate transaminase, and (iii) the correlation of metabolic responses to inhibitors with anti-proliferative effects. This demonstrates that the potential application of metabolic inhibitors necessitates personalized tumor diagnostics to identify individually optimal combinations of drugs. Furthermore, the investigation of these inhibitors in combination with chemotherapeutic agents or modulators of signaling pathways could possibly lead to optimized therapeutic concepts. Molekularbiologie und Tumorforschung Neumann HP, Pawlu C, Peczkowska M, Bausch B, McWhinney SR, Muresan M, Buchta M, Franke G, Klisch J, Bley TA et al. 2004 Distinct clinical features of paraganglioma syndromes associated with SDHB and SDHD gene mutations. Jama 292 943-951. (doi:10.1001/jama.292.8.943) Mi RK, Min SK, Ji EO, Yoo RK, Sang YS, Seong IS, Ji YL, Nam JY + Sug HL 2009 Mutational analysis of IDH1 codon 132 in glioblastomas and other common cancers. International Journal of Cancer 125 353-355. (doi:10.1002/ijc.24379) Cao W, Yacoub S, Shiverick KT, Namiki K, Sakai Y, Porvasnik S, Urbanek C + Rosser CJ 2008 Dichloroacetate (DCA) sensitizes both wild-type and over expressing bcl-2 prostate cancer cells in vitro to radiation. Prostate 68 1223-1231. (doi:10.1002/pros.20788) Paumen MB, Ishida Y, Han H, Muramatsu M, Eguchi Y, Tsujimoto Y + Honjo T 1997 Direct interaction of the mitochondrial membrane protein carnitine palmitoyltransferase I with Bcl-2. Biochemical and Biophysical Research Communications 231 523-525. (doi:10.1006/bbrc.1997.6089) Martinez-Zaguilan R, Seftor EA, Seftor RE, Chu YW, Gillies RJ + Hendrix MJ 1996 Acidic pH enhances the invasive behavior of human melanoma cells. Clin Exp Metastasis 14 176- 186. (doi:10.1007/BF00121214) Warburg O, Posener K + Negelein E 1924 Über den Stoffwechsel der Carcinomzelle. Biochemische Zeitschrift 152 309-344. (doi:10.1007/BF01058673) Roche TE + Hiromasa Y 2007 Pyruvate dehydrogenase kinase regulatory mechanisms and inhibition in treating diabetes, heart ischemia, and cancer. Cellular and Molecular Life Sciences 64 830-849. (doi:10.1007/s00018-007-6380-z) Brahimi-Horn MC, Chiche J + Pouysségur J 2007 Hypoxia and cancer. Journal of Molecular Medicine 85 1301-1307. (doi:10.1007/s00109-007-0281-3) Pinheiro C, Longatto-Filho A, Scapulatempo C, Ferreira L, Martins S, Pellerin L, Rodrigues M, Alves VAF, Schmitt F + Baltazar F 2008a Increased expression of monocarboxylate transporters 1, 2, and 4 in colorectal carcinomas. Virchows Archiv 452 139-146. (doi:10.1007/s00428-007-0558-5) Pawlu C, Bausch B + Neumann HPH 2005 Mutations of the SDHB and SDHD genes. Familial Cancer 4 49-54. (doi:10.1007/s10689-004-4227-4) Cohen AL, Holmen SL + Colman H 2013 IDH1 and IDH2 mutations in gliomas. Current Neurology and Neuroscience Reports 13 345. (doi:10.1007/s11910-013-0345-4) Lopaschuk GD, Belke DD, Gamble J, Itoi T + Schönekess BO 1994 Regulation of fatty acid oxidation in the mammalian heart in health and disease. Biochimica et Biophysica Acta 1213 263-276. (doi:10.1016/0005-2760(94)00082-4) Kruszynska YT + Sherratt HS 1987 Glucose kinetics during acute and chronic treatment of rats with 2[6(4-chloro-phenoxy)hexyl]oxirane-2-carboxylate, etomoxir. Biochemical Pharmacology 36 3917-3921. (doi:10.1016/0006-2952(87)90458-8) Rempel A, Mathupala SP + Pedersen PL 1996 Glucose catabolism in cancer cells: Regulation of the type II hexokinase promoter by glucose and cyclic AMP. FEBS Letters 385 233-237. (doi:10.1016/0014-5793(96)00399-7) Mosmann T 1983 Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65 55-63. (doi:10.1016/0022-1759(83)90303-4) Eboli ML 1985 Pyruvate dehydrogenase levels in Morris hepatomas with different growth rate. Cancer Letters 26 185-190. (doi:10.1016/0304-3835(85)90025-4) Marín-Hernández A, Gallardo-Pérez JC, Rodríguez-Enríquez S, Encalada R, Moreno-Sánchez R + Saavedra E 2011 Modeling cancer glycolysis. Biochimica et Biophysica Acta - Bioenergetics 1807 755-767. (doi:10.1016/j.bbabio.2010.11.006) Divakaruni AS, Rogers GW, Andreyev AY + Murphy AN 2016 The CPT inhibitor etomoxir has an off-target effect on the adenine nucleotide translocase and respiratory complex I. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1857 e118. (doi:10.1016/j.bbabio.2016.04.250) Kankotia S + Stacpoole PW 2014 Dichloroacetate and cancer: New home for an orphan drug? Biochimica et Biophysica Acta 1846 617-629. (doi:10.1016/j.bbcan.2014.08.005) Hatzivassiliou G, Zhao F, Bauer DE, Andreadis C, Shaw AN, Dhanak D, Hingorani SR, Tuveson DA + Thompson CB 2005 ATP citrate lyase inhibition can suppress tumor cell growth. Cancer Cell 8 311-321. (doi:10.1016/j.ccr.2005.09.008) Fantin VR, St-Pierre J + Leder P 2006 Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell 9 425-434. (doi:10.1016/j.ccr.2006.04.023) Bui T + Thompson CB 2006 Cancer's sweet tooth. Cancer Cell 9 419-420. (doi:10.1016/j.ccr.2006.05.012) Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T, Clark L, Bayani N, Coppe JP, Tong F et al. 2006 A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell 10 515-527. (doi:10.1016/j.ccr.2006.10.008) Guertin DA + Sabatini DM 2007 Defining the Role of mTOR in Cancer. Cancer Cell 12 9-22. (doi:10.1016/j.ccr.2007.05.008) Caro P, Kishan AU, Norberg E, Stanley IA, Chapuy B, Ficarro SB, Polak K, Tondera D, Gounarides J, Yin H et al. 2012 Metabolic Signatures Uncover Distinct Targets in Molecular Subsets of Diffuse Large B Cell Lymphoma. Cancer Cell 22 547-560. (doi:10.1016/j.ccr.2012.08.014) Bensaad K, Tsuruta A, Selak MA, Vidal MNC, Nakano K, Bartrons R, Gottlieb E + Vousden KH 2006 TIGAR, a p53-Inducible Regulator of Glycolysis and Apoptosis. Cell 126 107-120. (doi:10.1016/j.cell.2006.05.036) Hanahan D + Weinberg RA 2011 Hallmarks of cancer: The next generation. Cell 144 646-674. (doi:10.1016/j.cell.2011.02.013) Mizushima N + Komatsu M 2011 Autophagy: Renovation of cells and tissues. Cell 147 728-741. (doi:10.1016/j.cell.2011.10.026) Guzy RD, Hoyos B, Robin E, Chen H, Liu L, Mansfield KD, Simon MC, Hammerling U + Schumacker PT 2005 Mitochondrial complex III is required for hypoxia-induced ROS production and cellular oxygen sensing. Cell Metabolism 1 401-408. (doi:10.1016/j.cmet.2005.05.001) Papandreou I, Cairns RA, Fontana L, Lim AL + Denko NC 2006 HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption. Cell Metabolism 3 187-197. (doi:10.1016/j.cmet.2006.01.012) Kim JW, Tchernyshyov I, Semenza GL + Dang C V. 2006 HIF-1-mediated expression of pyruvate dehydrogenase kinase: A metabolic switch required for cellular adaptation to hypoxia. Cell Metabolism 3 177-185. (doi:10.1016/j.cmet.2006.02.002) Currie E, Schulze A, Zechner R, Walther TC + Farese R V. 2013 Cellular fatty acid metabolism and cancer. Cell Metabolism 18 153-161. (doi:10.1016/j.cmet.2013.05.017) 163 Mazan-Mamczarz K, Hagner P, Dai B, Corl S, Liu Z + Gartenhaus RB 2009 Targeted suppression of MCT-1 attenuates the malignant phenotype through a translational mechanism. Leukemia Research 33 474-482. (doi:10.1016/j.leukres.2008.08.012) Nagelkerke A, Sweep FCGJ, Geurts-Moespot A, Bussink J + Span PN 2015 Therapeutic targeting of autophagy in cancer. Part I: Molecular pathways controlling autophagy. Seminars in Cancer Biology 31 89-98. (doi:10.1016/j.semcancer.2014.05.004) Hill RP, Marie-Egyptienne DT + Hedley DW 2009 Cancer Stem Cells, Hypoxia and Metastasis. Seminars in Radiation Oncology 19 106-111. (doi:10.1016/j.semradonc.2008.12.002) Amelio I, Cutruzzolá F, Antonov A, Agostini M + Melino G 2014 Serine and glycine metabolism in cancer. Trends in Biochemical Sciences 39 191-198. (doi:10.1016/j.tibs.2014.02.004) Máximo V, Soares P, Lima J, Cameselle-Teijeiro J + Sobrinho-Simões M 2002 Mitochondrial DNA somatic mutations (point mutations and large deletions) and mitochondrial DNA variants in human thyroid pathology: a study with emphasis on Hürthle cell tumors. The American Journal of Pathology 160 1857-1865. (doi:10.1016/S0002-9440(10)61132-7) Gansler TS, Hardman W, Hunt DA, Schaffel S + Hennigar RA 1997 Increased expression of fatty acid synthase (OA-519) in ovarian neoplasms predicts shorter survival. Human Pathology 28 686-692. (doi:10.1016/S0046-8177(97)90177-5) Harris RA, Bowker-Kinley MM, Huang B + Wu P 2002 Regulation of the activity of the pyruvate dehydrogenase complex. In Advances in Enzyme Regulation, pp 249-259. (doi:10.1016/S0065-2571(01)00061-9) Rottem S + Naot Y 1998 Subversion and exploitation of host cells by mycoplasmas. Trends in Microbiology 6 436-440. (doi:10.1016/S0966-842X(98)01358-4) Pinheiro C, Sousa B, Albergaria A, Paredes J, Dufloth R, Vieira D, Schmitt F + Baltazar F 2011 GLUT1 and CAIX expression profiles in breast cancer correlate with adverse prognostic factors and MCT1 overexpression. Histology and Histopathology 26 1279-1286. (doi:10.1016/S1359-6349(10)71197-6) Pennacchietti S, Michieli P, Galluzzo M, Mazzone M, Giordano S + Comoglio PM 2003 Hypoxia promotes invasive growth by transcriptional activation of the met protooncogene. Cancer Cell 3 347-361. (doi:10.1016/S1535-6108(03)00085-0) Niemann S + Müller U 2000 Mutations in SDHC cause autosomal dominant paraganglioma, type 3. Nature Genetics 26 268-270. (doi:10.1038/81551) Lin G, Hill DK, Andrejeva G, Boult JKR, Troy H, Fong a-CLFWT, Orton MR, Panek R, Parkes HG, Jafar M et al. 2014 Dichloroacetate induces autophagy in colorectal cancer cells and tumours. British Journal of Cancer 111 375-385. (doi:10.1038/bjc.2014.281) Jang M, Kim SS + Lee J 2013 Cancer cell metabolism: implications for therapeutic targets. Experimental + Molecular Medicine 45 e45. (doi:10.1038/emm.2013.85) Christofk HR, Vander Heiden MG, Harris MH, Ramanathan A, Gerszten RE, Wei R, Fleming MD, Schreiber SL + Cantley LC 2008 The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature 452 230-233. (doi:10.1038/nature06734) Knobloch M, Braun SMG, Zurkirchen L, von Schoultz C, Zamboni N, Araúzo-Bravo MJ, Kovacs WJ, Karalay O, Suter U, Machado RAC et al. 2013 Metabolic control of adult neural stem cell activity by Fasn-dependent lipogenesis. Nature 493 226-230. (doi:10.1038/nature11689) Jiang P, Du W, Mancuso A, Wellen KE + Yang X 2013 Reciprocal regulation of p53 and malic enzymes modulates metabolism and senescence. Nature 493 689-693. (doi:10.1038/nature11776) Domcke S, Sinha R, Levine D a, Sander C + Schultz N 2013 Evaluating cell lines as tumour models by comparison of genomic profiles. Nature Communications 4 2126. (doi:10.1038/ncomms3126) Locasale JW, Grassian AR, Melman T, Lyssiotis CA, Mattaini KR, Bass AJ, Heffron G, Metallo CM, Muranen T, Sharfi H et al. 2011 Phosphoglycerate dehydrogenase diverts glycolytic flux and contributes to oncogenesis. Nature Genetics 43 869-874. (doi:10.1038/ng.890) Altman BJ, Stine ZE + Dang C V 2016 From Krebs to clinic: glutamine metabolism to cancer therapy. Nature Reviews Cancer 16 619-634. (doi:10.1038/nrc.2016.71) Gatenby RA + Gillies RJ 2004 Why do cancers have high aerobic glycolysis? Nature Reviews Cancer 4 891-899. (doi:10.1038/nrc1478) Menendez JA + Lupu R 2007 Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis. Nature Reviews. Cancer 7 763-777. (doi:10.1038/nrc2222) Dang C V, Kim J, Gao P + Yustein J 2008 The interplay between MYC and HIF in cancer. Nature Reviews Cancer 8 51-56. (doi:10.1038/nrc2274) Bristow RG + Hill RP 2008 Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability. Nature Reviews. Cancer 8 180-192. (doi:10.1038/nrc2344) Denko NC 2008 Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nature Reviews Cancer 8 705-713. (doi:10.1038/nrc2468) Cairns RA, Harris IS + Mak TW 2011 Regulation of cancer cell metabolism. Nature Reviews Cancer 11 85-95. (doi:10.1038/nrc2981) Carracedo A, Cantley LC + Pandolfi PP 2013 Cancer metabolism: fatty acid oxidation in the limelight. Nat.Rev.Cancer 13 227-232. (doi:10.1038/nrc3483.Cancer) Michelakis ED, Webster L + Mackey JR 2008 Dichloroacetate (DCA) as a potential metabolictargeting therapy for cancer. British Journal of Cancer 99 989-994. (doi:10.1038/sj.bjc.6604554) Giordano A, Calvani M, Petillo O, Grippo P, Tuccillo F, Melone M, Bonelli P, Calarco A + Peluso G 2005 tBid induces alterations of mitochondrial fatty acid oxidation flux by malonyl-CoAindependent inhibition of carnitine palmitoyltransferase-1. Cell Death and Differentiation 12 603-613. (doi:10.1038/sj.cdd.4401636) Mazurek S, Zwerschke W, Jansen-Dürr P + Eigenbrodt E 2001 Metabolic cooperation between different oncogenes during cell transformation: interaction between activated ras and HPV-16 E7. Oncogene 20 6891-6898. (doi:10.1038/sj.onc.1204792) Modena P, Testi MA, Facchinetti F, Mezzanzanica D, Radice MT, Pilotti S + Sozzi G 2003 UQCRH gene encoding mitochondrial Hinge protein is interrupted by a translocation in a softtissue sarcoma and epigenetically inactivated in some cancer cell lines. Oncogene 22 4586-4593. (doi:10.1038/sj.onc.1206472) Bandyopadhyay S, Pai SK, Watabe M, Gross SC, Hirota S, Hosobe S, Tsukada T, Miura K, Saito K, Markwell SJ et al. 2005 FAS expression inversely correlates with PTEN level in prostate cancer and a PI 3-kinase inhibitor synergizes with FAS siRNA to induce apoptosis. Oncogene 24 5389-5395. (doi:10.1038/sj.onc.1208555) Charafe-Jauffret E, Ginestier C, Monville F, Finetti P, Adélaïde J, Cervera N, Fekairi S, Xerri L, Jacquemier J, Birnbaum D et al. 2005 Gene expression profiling of breast cell lines identifies potential new basal markers. Oncogene 25 2273-2284. (doi:10.1038/sj.onc.1209254) Lin C-C, Cheng T-L, Tsai W-H, Tsai H-J, Hu K-H, Chang H-C, Yeh C-W, Chen Y-C, Liao C-C + Chang W-T 2012 Loss of the respiratory enzyme citrate synthase directly links the Warburg effect to tumor malignancy. Scientific Reports 2 785. (doi:10.1038/srep00785) Abramczyk H, Surmacki J, Kopeć M, Olejnik AK, Lubecka-Pietruszewska K + FabianowskaMajewska K 2015 The role of lipid droplets and adipocytes in cancer. Raman imaging of cell cultures: MCF10A, MCF7, and MDA-MB-231 compared to adipocytes in cancerous human breast tissue. The Analyst 140 2224-2235. (doi:10.1039/C4AN01875C) Halestrap AP + Price NT 1999 The proton-linked monocarboxylate transporter (MCT) family: structure, function and regulation. The Biochemical Journal 343 Pt 2 281-299. (doi:10.1042/0264-6021:3430281) Crabtree HG 1928 The carbohydrate metabolism of certain pathological overgrowths. Biochemical Journal 22 1289-1298. (doi:10.1042/bj0221289) Petros JA, Baumann AK, Ruiz-Pesini E, Amin MB, Sun CQ, Hall J, Lim S, Issa MM, Flanders WD, Hosseini SH et al. 2005 mtDNA mutations increase tumorigenicity in prostate cancer. Proceedings of the National Academy of Sciences of the United States of America 102 719-724. (doi:10.1073/pnas.0408894102) Launonen V, Vierimaa O, Kiuru M, Isola J, Roth S, Pukkala E, Sistonen P, Herva R + Aaltonen LA 2001 Inherited susceptibility to uterine leiomyomas and renal cell cancer. Proceedings of the National Academy of Sciences of the United States of America 98 3387-3392. (doi:10.1073/pnas.051633798) Cairns RA, Papandreou I, Sutphin PD + Denko NC 2007 Metabolic targeting of hypoxia and HIF1 in solid tumors can enhance cytotoxic chemotherapy. Proceedings of the National Academy of Sciences 104 9445-9450. (doi:10.1073/pnas.0611662104) DeBerardinis RJ, Mancuso A, Daikhin E, Nissim I, Yudkoff M, Wehrli S + Thompson CB 2007 Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis. Proceedings of the National Academy of Sciences of the United States of America 104 19345-19350. (doi:10.1073/pnas.0709747104) Khasawneh J, Schulz MD, Walch A, Rozman J, Hrabe de Angelis M, Klingenspor M, Buck A, Schwaiger M, Saur D, Schmid RM et al. 2009 Inflammation and mitochondrial fatty acid beta-oxidation link obesity to early tumor promotion. Proceedings of the National Academy of Sciences of the United States of America 106 3354-3359. (doi:10.1073/pnas.0802864106) Guillaumond F, Bidaut G, Ouaissi M, Servais S, Gouirand V, Olivares O, Lac S, Borge L, Roques J, Gayet O et al. 2015 Cholesterol uptake disruption, in association with chemotherapy, is a promising combined metabolic therapy for pancreatic adenocarcinoma. Proceedings of the National Academy of Sciences of the United States of America 112 2473-2478. (doi:10.1073/pnas.1421601112) Duranteau J, Chandel NS, Kulisz A, Shao Z + Schumacker PT 1998 Intracellular signaling by reactive oxygen species during hypoxia in cardiomyocytes. Journal of Biological Chemistry 273 11619-11624. (doi:10.1074/jbc.273.19.11619) Mathupala SP, Rempel A + Pedersen PL 2001 Glucose catabolism in cancer cells: Identification and characterization of a marked activation response of the type II hexokinase gene to hypoxic conditions. Journal of Biological Chemistry 276 43407-43412. (doi:10.1074/jbc.M108181200) McFate T, Mohyeldin A, Lu H, Thakar J, Henriques J, Halim ND, Wu H, Schell MJ, Tsang TM, Teahan O et al. 2008 Pyruvate dehydrogenase complex activity controls metabolic and malignant phenotype in cancer cells. The Journal of Biological Chemistry 283 22700- 22708. (doi:10.1074/jbc.M801765200) Astuti D, Latif F, Dallol a, Dahia PL, Douglas F, George E, Sköldberg F, Husebye ES, Eng C + Maher ER 2001 Gene mutations in the succinate dehydrogenase subunit SDHB cause susceptibility to familial pheochromocytoma and to familial paraganglioma. American Journal of Human Genetics 69 49-54. (doi:10.1086/321282) Colombo N, Peiretti M, Parma G, Lapresa M, Mancari R, Carinelli S, Sessa C + Castiglione M 2010 Newly diagnosed and relapsed epithelial ovarian carcinoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Annals of Oncology 21 23-30. (doi:10.1093/annonc/mdq244) Ledermann JA, Raja FA, Fotopoulou C, Gonzalez-Martin A, Colombo N + Sessa C 2013 Newly diagnosed and relapsed epithelial ovarian carcinoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Annals of Oncology 24. (doi:10.1093/annonc/mdt333) Loda M, Migita T, Ruiz S, Fornari A, Fiorentino M, Priolo C, Zadra G, Inazuka F, Grisanzio C, Palescandolo E et al. 2009 Fatty acid synthase: A metabolic enzyme and candidate oncogene in prostate cancer. Journal of the National Cancer Institute 101 519-532. (doi:10.1093/jnci/djp030) Kurman RJ + Shih I-M 2010 The origin and pathogenesis of epithelial ovarian cancer: a proposed unifying theory. The American Journal of Surgical Pathology 34 433-443. (doi:10.1097/PAS.0b013e3181cf3d79) Pinheiro C, Longatto-Filho A, Ferreira L, Pereira SMM, Etlinger D, Moreira M a R, Jubé LF, Queiroz GS, Schmitt F + Baltazar F 2008b Increasing expression of monocarboxylate transporters 1 and 4 along progression to invasive cervical carcinoma. International Journal of Gynecological Pathology: Official Journal of the International Society of Gynecological Pathologists 27 568-574. (doi:10.1097/PGP.0b013e31817b5b40) Grassian AR, Metallo CM, Coloff JL, Stephanopoulos G + Brugge JS 2011 Erk regulation of pyruvate dehydrogenase flux through PDK4 modulates cell proliferation. Genes and Development 25 1716-1733. (doi:10.1101/gad.16771811) Baysal BE, Willett-Brozick JE, Lawrence EC, Drovdlic CM, Savul SA, McLeod DR, Yee HA, Brackmann DE, Slattery WH, Myers EN et al. 2002 Prevalence of SDHB, SDHC, and SDHD germline mutations in clinic patients with head and neck paragangliomas. Journal of Medical Genetics 39 178-183. (doi:10.1111/j.1399-0004.2004.00328.x) Chiche J, Brahimi-Horn MC + Pouysségur J 2010 Tumour hypoxia induces a metabolic shift causing acidosis: A common feature in cancer. Journal of Cellular and Molecular Medicine 14 771-794. (doi:10.1111/j.1582-4934.2009.00994.x) Pfeiffer T, Schuster S, Bonhoeffer S, Angulo-Brown F, Santillan M, Calleja-Quevedo E, Waddell TG, Repovic P, Melendez-Hevia E, Heinrich R et al. 2001 Cooperation and competition in the evolution of ATP-producing pathways. Science (New York, N.Y.) 292 504-507. (doi:10.1126/science.1058079) Matoba S, Kang J-G, Patino WD, Wragg A, Boehm M, Gavrilova O, Hurley PJ, Bunz F + Hwang PM 2006 P53 Regulates Mitochondrial Respiration. Science 312 1650-1653. (doi:10.1126/science.1126863) Vander Heiden MG, Cantley LC + Thompson CB 2009 Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science (New York, N.Y.) 324 1029-1033. (doi:10.1126/science.1160809) Parsons DW, Jones S, Zhang X, Lin JC-H, Leary RJ, Angenendt P, Mankoo P, Carter H, Siu I-M, Gallia GL et al. 2008 An integrated genomic analysis of human glioblastoma multiforme. Science 321 1807-1812. (doi:10.1126/science.1164382) Bricker DK, Taylor EB, Schell JC, Orsak T, Boutron A, Chen Y-C, Cox JE, Cardon CM, Van Vranken JG, Dephoure N et al. 2012 A mitochondrial pyruvate carrier required for pyruvate uptake in yeast, Drosophila, and humans. Science 337 96-100. (doi:10.1126/science.1218099) Herzig S, Raemy E, Montessuit S, Veuthey J-L, Zamboni N, Westermann B, Kunji ERS, Martinou J-C, Hiltunen JK, Chen Z et al. 2012 Identification and functional expression of the mitochondrial pyruvate carrier. Science (New York, N.Y.) 337 93-96. (doi:10.1126/science.1218530) Baysal BE, Ferrell RE, Willett-Brozick JE, Lawrence EC, Myssiorek D, Bosch a, van der Mey a, Taschner PE, Rubinstein WS, Myers EN et al. 2000 Mutations in SDHD, a mitochondrial complex II gene, in hereditary paraganglioma. Science (New York, NY) 287 848-851. (doi:10.1126/science.287.5454.848) Loftus TM, Jaworsky DE, Frehywot GL, Townsend C a, Ronnett G V, Lane MD + Kuhajda FP 2000 Reduced food intake and body weight in mice treated with fatty acid synthase inhibitors. Science (New York, N.Y.) 288 2379-2381. (doi:10.1126/science.288.5475.2379) Kim J, Gao P, Liu Y-C, Semenza GL + Dang C V 2007 Hypoxia-inducible factor 1 and dysregulated c-Myc cooperatively induce vascular endothelial growth factor and metabolic switches hexokinase 2 and pyruvate dehydrogenase kinase 1. Molecular and Cellular Biology 27 7381-7393. (doi:10.1128/MCB.00440-07) Kamarajugadda S, Stemboroski L, Cai Q, Simpson NE, Nayak S, Tan M + Lu J 2012 Glucose Oxidation Modulates Anoikis and Tumor Metastasis. Molecular and Cellular Biology 32 1893-1907. (doi:10.1128/MCB.06248-11) Erler JT, Cawthorne CJ, Williams KJ, Koritzinsky M, Wouters BG, Wilson C, Miller C, Demonacos C, Stratford IJ + Dive C 2004 Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and -independent mechanisms and contributes to drug resistance. Molecular and Cellular Biology 24 2875-2889. (doi:10.1128/MCB.24.7.2875-2889.2004) Warburg O 1956 Origin of cancer cells. Oncologia 9 75-83. (doi:10.1136/bmj.1.4082.694-a) Lehtonen HJ, Kiuru M, Ylisaukko-Oja SK, Salovaara R, Herva R, Koivisto PA, Vierimaa O, Aittomäki K, Pukkala E, Launonen V et al. 2006 Increased risk of cancer in patients with fumarate hydratase germline mutation. Journal of Medical Genetics 43 523-526. (doi:10.1136/jmg.2005.036400) Czernin J + Phelps ME 2002 Positron emission tomography scanning: current and future applications. Annual Review of Medicine 53 89-112. (doi:10.1146/annurev.med.53.082901.104028) Cardaci S + Ciriolo MR 2012 TCA cycle defects and cancer: When metabolism tunes redox state. International Journal of Cell Biology. (doi:10.1155/2012/161837) Chajès V, Cambot M, Moreau K, Lenoir GM + Joulin V 2006 Acetyl-CoA carboxylase alpha is essential to breast cancer cell survival. Cancer Research 66 5287-5294. (doi:10.1158/0008-5472.CAN-05-1489) Rofstad EK, Mathiesen B, Kindem K + Galappathi K 2006 Acidic extracellular pH promotes experimental metastasis of human melanoma cells in athymic nude mice. Cancer Research 66 6699-6707. (doi:10.1158/0008-5472.CAN-06-0983) Accioly MT, Pacheco P, Maya-Monteiro CM, Carrossini N, Robbs BK, Oliveira SS, Kaufmann C, Morgado-Diaz JA, Bozza PT + Viola JPB 2008 Lipid bodies are reservoirs of cyclooxygenase-2 and sites of prostaglandin-E2 synthesis in colon cancer cells. Cancer Research 68 1732-1740. (doi:10.1158/0008-5472.CAN-07-1999) Hirschhaeuser F, Sattler UGA + Mueller-Klieser W 2011 Lactate: A metabolic key player in cancer. Cancer Research 71 6921-6925. (doi:10.1158/0008-5472.CAN-11-1457) Robey IF, Baggett BK, Kirkpatrick ND, Roe DJ, Dosescu J, Sloane BF, Hashini AI, Morse DL, Raghunand N, Gatenby RA et al. 2009 Bicarbonate increases tumor pH and inhibits spontaneous metastases. Cancer Research 69 2260-2268. (doi:10.1158/0008-5472.CAN07-5575) Migita T, Narita T, Nomura K, Miyagi E, Inazuka F, Matsuura M, Ushijima M, Mashima T, Seimiya H, Satoh Y et al. 2008 ATP citrate lyase: Activation and therapeutic implications in non-small cell lung cancer. Cancer Research 68 8547-8554. (doi:10.1158/0008-5472.CAN08-1235) Chang Q, Jurisica I, Do T + Hedley DW 2011 Hypoxia predicts aggressive growth and spontaneous metastasis formation from orthotopically grown primary xenografts of human pancreatic cancer. Cancer Research 71 3110-3120. (doi:10.1158/0008-5472.CAN10-4049) Palmieri D, Fitzgerald D, Shreeve SM, Hua E, Bronder JL, Weil RJ, Davis S, Stark AM, Merino MJ, Kurek R et al. 2009 Analyses of resected human brain metastases of breast cancer reveal the association between up-regulation of hexokinase 2 and poor prognosis. Molecular Cancer Research 7 1438-1445. (doi:10.1158/1541-7786.mcr-09-0234) Qiu B, Ackerman D, Sanchez DJ, Li B, Ochocki JD, Grazioli A, Bobrovnikova-Marjon E, Alan Diehl J, Keith B + Celeste Simon M 2016 HIF2alpha-dependent lipid storage promotes endoplasmic reticulum homeostasis in clear-cell renal cell carcinoma. Cancer Discovery 5 653-667. (doi:10.1158/2159-8290.CD-14-1507) Fiume L, Manerba M, Vettraino M + Di Stefano G 2010 Impairment of aerobic glycolysis by inhibitors of lactic dehydrogenase hinders the growth of human hepatocellular carcinoma cell lines. Pharmacology 86 157-162. (doi:10.1159/000317519) Rouschop KM, van den Beucken T, Dubois L, Niessen H, Bussink J, Savelkouls K, Keulers T, Mujcic H, Landuyt W, Voncken JW et al. 2010 The unfolded protein response protects human tumor cells during hypoxia through regulation of the autophagy genes MAP1LC3B and ATG5. J Clin Invest 120 127-141. (doi:10.1172/jci40027) Kioi M, Vogel H, Schultz G, Hoffman RM, Harsh GR + Brown JM 2010 Inhibition of vasculogenesis, but not angiogenesis, prevents the recurrence of glioblastoma after irradiation in mice. Journal of Clinical Investigation 120 694-705. (doi:10.1172/JCI40283) Doherty JR + Cleveland JL 2013 Targeting lactate metabolism for cancer therapeutics. The Journal of Clinical Investigation 123 3685-3692. (doi:10.1172/JCI69741) Cairns RA, Iqbal J, Lemonnier F, Kucuk C, De Leval L, Jais JP, Parrens M, Martin A, Xerri L, Brousset P et al. 2012 IDH2 mutations are frequent in angioimmunoblastic T-cell lymphoma. Blood 119 1901-1903. (doi:10.1182/blood-2011-11-391748) 162 Pértega-Gomes N, Vizcaíno JR, Miranda-Gonçalves V, Pinheiro C, Silva J, Pereira H, Monteiro P, Henrique RM, Reis RM, Lopes C et al. 2011 Monocarboxylate transporter 4 (MCT4) and CD147 overexpression is associated with poor prognosis in prostate cancer. BMC Cancer 11 312. (doi:10.1186/1471-2407-11-312) Carew JS + Huang P 2002 Mitochondrial defects in cancer. Molecular Cancer 1 9. (doi:10.1186/1476-4598-1-9) Anso E, Mullen AR, Felsher DW, Matés JM, Deberardinis RJ + Chandel NS 2013 Metabolic changes in cancer cells upon suppression of MYC. Cancer + Metabolism 1 7. (doi:10.1186/2049-3002-1-7) de Gonzalo-Calvo D, López-Vilaró L, Nasarre L, Perez-Olabarria M, Vázquez T, Escuin D, Badimon L, Barnadas A, Lerma E + Llorente-Cortés V 2015 Intratumor cholesteryl ester accumulation is associated with human breast cancer proliferation and aggressive potential: a molecular and clinicopathological study. BMC Cancer 15 460. (doi:10.1186/s12885-015-1469-5) Reinartz S, Finkernagel F, Adhikary T, Rohnalter V, Schumann T, Schober Y, Nockher WA, Nist A, Stiewe T, Jansen JM et al. 2016 A transcriptome-based global map of signaling pathways in the ovarian cancer microenvironment associated with clinical outcome. Genome Biology 17 108. (doi:10.1186/s13059-016-0956-6) Carvajal-Carmona LG, Alam NA, Pollard PJ, Jones AM, Barclay E, Wortham N, Pignatelli M, Freeman A, Pomplun S, Ellis I et al. 2006 Adult leydig cell tumors of the testis caused by germline fumarate hydratase mutations. Journal of Clinical Endocrinology and Metabolism 91 3071-3075. (doi:10.1210/jc.2006-0183) Owens KM, Kulawiec M, Desouki MM, Vanniarajan A + Singh KK 2011 Impaired OXPHOS complex III in breast cancer. PLoS ONE 6. (doi:10.1371/journal.pone.0023846) Miskimins WK, Ahn HJ, Kim JY, Ryu S, Jung YS + Choi JY 2014 Synergistic anti-cancer effect of phenformin and oxamate. PLoS ONE 9. (doi:10.1371/journal.pone.0085576) Qu Y, Han B, Yu Y, Yao W, Bose S, Karlan BY, Giuliano AE + Cui X 2015 Evaluation of MCF10A as a reliable model for normal human mammary epithelial cells. PLoS ONE 10. (doi:10.1371/journal.pone.0131285) Rohnalter V, Roth K, Finkernagel F, Adhikary T, Obert J, Dorzweiler K, Bensberg M, MullerBrusselbach S + Muller R 2015 A multi-stage process including transient polyploidization and EMT precedes the emergence of chemoresistent ovarian carcinoma cells with a dedifferentiated and pro-inflammatory secretory phenotype. Oncotarget 6 40005-40025. (doi:10.18632/oncotarget.5552) Lengyel E 2010 Ovarian cancer development and metastasis. The American Journal of Pathology 177 1053-1064. (doi:10.2353/ajpath.2010.100105) Hamilton E, Fennell M + Stafford DM 1995 Modification of tumour glucose metabolism for therapeutic benefit. Acta Oncol 34 429-433. (doi:10.3109/02841869509094003) Jeoung NH + Harris RA 2010 Role of pyruvate dehydrogenase kinase 4 in regulation of blood glucose levels. Korean Diabetes Journal 34 274-283. (doi:10.4093/kdj.2010.34.5.274) Van Kuppeveld FJM, Van der Logt JTM, Angulo AF, Van Zoest MJ, Quint WG V, Niesters HGM, Galama JMD + Melchers WJG 1992 Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification. Applied and Environmental Microbiology 58 2606-2615. (doi:1381174) Saddik M, Gamble J, Witters LA + Lopaschuk GD 1993 Acetyl-CoA carboxylase regulation of fatty acid oxidation in the heart. The Journal of Biological Chemistry 268 25836-25845. Drapier J-C + Hibbs JB 1996 [3] Aconitases: A class of metalloproteins highly sensitive to nitric oxide synthesis. Methods in Enzymology 269 26-36. (doi:10.1016/S0076-6879(96)69006- 5) Kauppinen RA, Sihra TS + Nicholls DG 1987 Aminooxyacetic acid inhibits the malate-aspartate shuttle in isolated nerve terminals and prevents the mitochondria from utilizing glycolytic substrates. BBA - Molecular Cell Research 930 173-178. (doi:10.1016/0167- 4889(87)90029-2) Locasale JW, Grassian A, Beroukhim R, Meyerson M, Wagner G, Asara JM, Brugge JS, Vander Heiden MG + Cantley LC 2012 Amplification of phosphoglycerate dehydrogenase diverts glycolytic flux and contributes to oncogenesis. BMC Proceedings 6. Bénard J, Da Silva J, De Blois MC, Boyer P, Riou G, Duvillard P + Chiric E 1985 Characterization of a Human Ovarian Adenocarcinoma Line, IGROV1, in Tissue Culture and in Nude Mice. Cancer Research 45 4970-4979. Hamilton TC, Young RC, McKoy WM, Grotzinger KR, Green JA, Chu EW, Whang-Peng J, Rogan AM, Green WR + Ozols RF 1983 Characterization of a human ovarian carcinoma cell line (NIH:OVCAR-3) with androgen and estrogen receptors. Cancer Research 43 5379-5389. Kornacker MS + Ball EC 1965 Citrate Cleavage in Adipose Tissue. Proceedings of the National Academy of Sciences 54 899-904. Buick RN, Pullano R + Trent JM 1985 Comparative Properties of Five Human Ovarian Adenocarcinoma Cell Lines. Cancer Research 45 3668-3676. Harris R, Huang B + Wu P 2001 Control of pyruvate dehydrogenase kinase gene expression. Advances in Enzyme Regulation 41 269-288. Jones JB, Song JJ, Hempen PM, Parmigiani G, Hruban RH + Kern SE 2001 Detection of mitochondrial DNA mutations in pancreatic cancer offers a 'mass'-ive advantage over detection of nuclear DNA mutations. Cancer Research 61 1299-1304. Eigenbrodt E, Reinacher M, Scheefers-Borchel U, Scheefers H + Friis R 1992 Double role for pyruvate kinase type M2 in the expansion of phosphometabolite pools found in tumor cells. Critical Reviews in Oncogenesis 3 91-115. Elwood JC 1968 Effect of Oxamate on Glycolysis and Respiration in Sarcoma 37 Ascites Cells. Cancer Research 28 2056-2060. Parolin MLML, Spriet LLLL, Hultman EE, Matsos MPMP, Hollidge-Horvat MGMG, Jones NLNL + Heigenhauser GJGJ 2000 Effects of PDH activation by dichloroacetate in human skeletal muscle during exercise in hypoxia. American Journal of Physiology-Endocrinology and Metabolism 279 E752-E761. Moreno-Sánchez R, Rodríguez-Enríquez S, Marín-Hernández A + Saavedra E 2007 Energy metabolism in tumor cells. The FEBS Journal 274 1393-1418. (doi:10.1111/j.1742- 4658.2007.05686.x) Eboli ML + Galeotti T 1981 Evidence for the occurrence of the malate-citrate shuttle in intact Ehrlich ascites tumor cells. BBA - Bioenergetics 638 75-79. (doi:10.1016/0005- 2728(81)90187-0) Barron C, Tsiani E + Tsakiridis T 2012 Expression of the glucose transporters GLUT1 , GLUT3 , GLUT4 and GLUT12 in human cancer cells. BMC Proceedings 6 P4. (doi:10.1186/1753- 6561-6-S3-P4) Kuhajda FP 2000 Fatty-acid synthase and human cancer: New perspectives on its role in tumor biology. Nutrition 16 202-208. Liu VW, Shi HH, Cheung a N, Chiu PM, Leung TW, Nagley P, Wong LC + Ngan HY 2001 High incidence of somatic mitochondrial DNA mutations in human ovarian carcinomas. Cancer Research 61 5998-6001. Fukumura D, Xu L, Chen Y, Gohongi T, Seed B + Jain RK 2001 Hypoxia and acidosis independently up-regulate vascular endothelial growth factor transcription in brain tumors in vivo. Cancer Research 61 6020-6024. Greenhouse WV V + Lehninger AL 1976 Occurrence of the Malate-Aspartate Shuttle in Various Tumor Types. Cancer Research 36 1392-1396. Awan MM + Saggerson ED 1993 Malonyl-CoA metabolism in cardiac myocytes and its relevance to the control of fatty acid oxidation. The Biochemical Journal 295 (Pt 1) 61-66. Hausladen A + Fridovich I 1996 Measuring nitric oxide and superoxide: rate constants for aconitase reactivity. Methods in Enzymology 269 37-41. Rose IA + O'Connell EL 1967 Mechanism of aconitase action. I. The hydrogen transfer reaction. Journal of Biological Chemistry 242 1870-1879. Coy JF, Dressler D, Wilde J + Schubert P 2005 Mutations in the transketolase-like gene TKTL1: Clinical implications for neurodegenerative diseases, diabetes and cancer. Clinical Laboratory 51 257-273. Halestrap A + P 1975 The mitochondrial pyruvate carrier. Kinetics and specificity for substrates and inhibitors. The Biochemical Journal 148 85-96. Hatefi Y + Stiggall DL 1978 Preparation and properties of succinate: Ubiquinone oxidoreductase (complex II). In Biomembranes - Part D: Biological Oxidations, pp 21-27. Ed SF and LPBT-M in Enzymology. Academic Press. (doi:http://dx.doi.org/10.1016/S0076- 6879(78)53008-5) Hansford RG + Cohen L 1978 Relative importance of pyruvate dehydrogenase interconversion and feed-back inhibition in the effect of fatty acids on pyruvate oxidation by rat heart mitochondria. Archives of Biochemistry and Biophysics 191 65-81. (doi:10.1016/0003- 9861(78)90068-1) Hakala MT, Glaid AJ + Schwert GW 1955 Lactic Dehydrogenase - Variation of kinetic and equilibrium constants with temperature. Journal of Biological Chemistry 221 191-209. Papaconstantinou J + Colowick SP 1961 The role of glycolysis in the growth of tumor cells. II. The effect of oxamic acid on the growth of HeLa cells in tissue culture. Journal of Biological Chemistry 236 285-288. Halestrap a P + Denton RM 1975 The specificity and metabolic implications of the inhibition of pyruvate transport in isolated mitochondria and intact tissue preparations by alphaCyano-4-hydroxycinnamate and related compounds. The Biochemical Journal 148 97- 106. Bakker EP + Van Dam K 1974 The movement of monocarboxylic acids across phospholipid membranes: evidence for an exchange diffusion between pyruvate and other monocarboxylate ions. BBA - Biomembranes 339 285-289. (doi:10.1016/0005- 2736(74)90325-3) Ein typisches Merkmal von Tumorzellen ist ihr veränderter Intermediärstoffwechsel. Tumor¬zellen sind häufig gekennzeichnet durch eine gesteigerte Glukoseaufnahme sowie die Bildung von Laktat auch in Anwesenheit von Sauerstoff (aerobe Glykolyse, "Warburg-Effekt"). Als weitere verstärkt ablaufende Stoffwechselwege rücken zunehmend der Abbau von Glutamin als anaplerotische Reaktion des Citratzyklus ("Glutaminolyse"), die de novo Fettsäuresynthese und der Fettsäureabbau über die β-Oxidation in den Mittelpunkt der Forschung. Die Inhibition dieser Stoffwechselwege könnte eine geeignete Angriffsstelle für künftige Krebstherapien sein. Das Ovarialkarzinom ist die gynäkologische Krebserkrankung mit der höchsten Letalität und weist häufig die Bildung von Aszites als besonderes Charakteristikum auf. Die Isolierung primärer Tumorzellen aus Aszites bietet eine einzigartige Möglichkeit zur Erforschung metabolischer Aberrationen in dieser Tumorentität. Ziel dieser Arbeit war die Identifizierung tumorspezifischer Stoffwechselwege als mögliche Angriffspunkte neuer Therapieansätze. Hierzu wurden patientenabgeleitete Ovarialkarzinomzellen im Vergleich zu einer Reihe etablierter Tumorzelllinien und der nicht-transformierten Brustepithelzelllinie MCF 10A eingesetzt. Voraussetzung dafür war die genaue Charakterisierung des metabolischen Phänotyps mit Hilfe von Expressionsanalysen und "metabolic profiling" mittels des Seahorse XFe Analyzers. Dafür wurden die metabolischen Parameter OCR (oxygen consumption rate) und ECAR (extracellular acidification rate) sowie die entsprechenden Kapazitäten unter Normalbedingungen und unter Anwendung von metabolischem Druck, vor allem durch den Einsatz von metabolischen Inhibitoren, gemessen. Im Zuge dieser Arbeit fiel eine Besonderheit im Stoffwechsel der Ovarialkarzinomzelllinie SKOV-3 auf. Die ATP-Produktion aus Glukose mittels oxidativer Phosphorylierung (OXPHOS) ist in diesen Zellen blockiert. Vermutet wurde zunächst ein Defekt im Citratzyklus. Dies ist jedoch nicht kompatibel mit dem Befund, dass SKOV-3 Zellen die ATP-Gewinnung mittels OXPHOS über einen längeren Zeitraum aufrechterhalten können. Deshalb wurde ein "fatty acid cycle" (FAC) als alternativer Stoffwechselweg postuliert, der aus dem zyklischen Auf- und Abbau von Fettsäuren besteht. Die Vorteile des FAC für die Tumorzelle könnten in einer Unabhängigkeit von externer Substratzufuhr bei einer geleichzeitig gesteigerten anabolen Kapazität aufgrund der Verlagerung des Glukosestoffwechsels in Richtung Pentosephosphatweg sein. Aufgrund der potentiellen Vorteile des FAC für proliferierende Zellen ist es wahrscheinlich, dass dieser Zyklus auch in anderen Zellen operativ ist. Die Ursache für die Besonderheiten des SKOV-3 Stoffwechsels ist möglicherweise ein verminderter Transport von Pyruvat in die Mitochondrienmatrix aufgrund einer verringerten Expression des Gens für den Mitochondrial Pyruvate Carrier 1 (MPC1). Ein weiterer wichtiger Befund war die Feststellung einer gesteigerten aeroben Glykolyse und β-Oxidation in allen patientenabgeleiteten Ovarialkarzinomzellen. Durch den Einsatz von Inhibitoren für Tumorzellen relevante Stoffwechselwege wurde die Möglichkeit einer Interferenz mit der Proliferation bzw. dem Überleben dieser Zellen mit Hilfe des xCELLigence RTCA untersucht. Dabei zeigte sich, dass einzelne Inhibitoren nur in exzessiv hohen Konzentrationen die Proliferation (wahrscheinlich unspezifisch) hemmten, dass aber die Kombination verschiedener Stoffwechselmodulatoren in Konzentrationen, in denen sie als einzelner Wirkstoff wirkungslos waren, zu deutlichen synergistischen Effekten führte. Vor allem die Kombination von Oxamat (Laktat-Dehydrogenase-Inhibitor) mit den Inhibitoren DCA (Inhibitor der Pyruvat-Dehydrogenase-Kinasen; PDKs), AOA (Inhibitor der Aspartat-Transaminase und damit des Malat-Aspartat-Shuttles) sowie SB204990 (Inhibitor der ATP-Citrat-Lyase) verursachte eine deutliche Proliferationsinhibition, bis hin zum Absterben der Zellen. Dennoch unterscheiden sich Ovarialkarzinomzellen von verschiedenen Patientinnen zum Teil deutlich hinsichtlich ihres Metabolismus bzw. der Auswirkungen von Inhibitoren auf die Proliferation. Beispiele hierfür sind Unterschiede bei (i) der Nutzung von Glutamin und Pyruvat als alleinigem Substrat, (ii) der Abhängigkeit von PDKs, Laktat-Dehydrogenase und Aspartat-Transaminase (iii) der Korrelation metabolischer Reaktionen auf Inhibitoren mit Effekten auf die Proliferation. Dies zeigt, dass eine potentielle Anwendung metabolischer Inhibitoren eine personalisierte Tumordiagnostik notwendig macht, um individuell optimale Kombinationen von Wirkstoffen zu identifizieren. Zudem könnte die Untersuchung dieser Hemmstoffe in Kombination mit Chemotherapeutika oder Modulatoren von Signalwegen ggf. zu optimierten Therapiekonzepten führen. Medizin doctoralThesis https://archiv.ub.uni-marburg.de/diss/z2017/0200/cover.png Obert, Julia Obert Julia Identifizierung von Stoffwechselwegen in humanen Ovarialkarzinomzellen mit potentiell therapeutischer Relevanz metabolism monograph 2017-03-16 ECAR proliferation Metabolismus 2017-03-03 urn:nbn:de:hebis:04-z2017-02002 Eierstockkrebs Proliferation ovarian cancer