Zusammenfassung:
Das duktale Adenokarzinom des Pankreas (PDAC) zeichnet sich durch eine hohe Mortalität aus: die Fünf-Jahres-Überlebensrate liegt bei nur 8%. Ursächlich hierfür ist zum einen die späte Diagnose aufgrund fehlender Frühsymptome, sodass der Tumor bei Erstdiagnose oftmals schon metastasiert und eine operative Entfernung nicht möglich ist. Zum anderen zeigen sich Pankreaskarzinome äußerst resistent gegen eine chemotherapeutische Behandlung.
Nach dem derzeitigen Stand der Forschung entsteht das PDAC aus Vorläuferläsionen durch eine Akkumulation charakteristischer Genmutationen. Die genauen molekularpathologischen Mechanismen sind jedoch noch nicht verstanden. Dementsprechend wurde in den letzten Jahren intensiv nach Proteinen gesucht, die eine wichtige Rolle in der Pathogenese des Pankreaskarzinoms spielen und somit Angriffspunkte für neue Therapien darstellen. Eines dieser Proteine ist die Branched-Chain α-Ketoacid Dehydrogenase-Kinase (BCKDK). In einem Kinom-Screening zeigte sich nach Repression von BCKDK in Pankreaskarzinomzellen eine erhöhte Apoptoserate. Zudem konnte eine vermehrte Expression des BCKDK-Gens in primären humanen Pankreaskarzinomgeweben nachgewiesen werden. Die BCKDK reguliert die Branched-Chain α-Ketoacid Dehydrogenase und ist somit ein Schlüsselenzym im Abbau verzweigtkettiger Aminosäuren. Eine weitere Funktion der Kinase ist bisher nicht beschrieben.
Im Rahmen dieser Arbeit wurde die möglicherweise pro-onkogene Wirkung der BCKDK im PDAC näher untersucht. Hierzu wurde die Expression der Kinase mithilfe dreier siRNAs (siRNA 1, 3 und 5) in zwei Pankreaskarzinomzelllinien supprimiert und jeweils die Auswirkungen auf Apoptose, Proliferation und Expression von Zielproteinen untersucht.
Die Versuchsreihen lieferten unterschiedliche Ergebnisse. Die Verwendung der siRNA 1 führte in beiden Zelllinien zu einer Abnahme der Zellviabilität. Als Ursache hierfür konnte eine Apoptoseinduktion nachgewiesen werden. Zusätzlich sank die Proliferationsrate und es kam zu einem Zellzyklusarrest der Zellen in der G2-Phase. Die Repression des Zielgens mit der siRNA 3 führte ebenfalls zu einer Abnahme der Zellviabilität durch Apoptoseinduktion, es kam jedoch nur in einer der verwendeten Zelllinien zu einer signifikanten Abnahme der Proliferationsrate. Die Zellen, die mit der siRNA 5 transfiziert worden waren, blieben hingegen sowohl in der Zellviabilität als auch in der Proliferation im Vergleich zur Kontrolle unverändert. Diese sehr unterschiedlichen Versuchsergebnisse sind möglicherweise durch Off-Target-Effekte der siRNAs zu erklären. Die zugrundeliegenden molekularpathologischen Mechanismen konnten in der vorliegenden Arbeit jedoch nicht vollständig aufgedeckt werden und bedürfen weiterer Analysen.
Bibliographie / References
- Raab RM, Stephanopoulos G (2004): Dynamics of gene silencing by RNA in- terference. In: Biotechnology and Bioengineering 88, S. 121-132.
- Real FX (2003): A "catastrophic hypothesis" for pancreas cancer progression. In: Gastroenterology 124, S. 1958-1964.
- Qiu S, Adema CM, Lane T (2005): A computational study of off-target ef- fects of RNA interference. In: Nucleic Acids Research 33, S. 1834-1847.
- Hutvágner G, Zamore PD (2002): A microRNA in a multiple-turnover RNAi enzyme complex. In: Science 297, S. 2056-2060.
- Suryawan A, Hawes JW, Harris RA et al. (1998): A molecular model of hu- man branched-chain amino acid metabolism. In: American Journal of Clinical Nutrition 68, S. 72-81.
- Tada M, Ohashi M, Shiratori Y et al. (1996): Analysis of K-ras gene mutation in hyperplastic duct cells of the pancreas without pancreatic disease. In: Gas- troenterology 110, S. 227-231.
- Harris RA, Popov KM, Zhao Y et al. (1995): A new family of protein kinases -the mitochondrial protein kinases. In: Advances in Enzyme Regulation 35, S. 147-162.
- Hruban RH, Takaori K, Klimstra DS et al. (2004): An illustrated consensus on the classification of pancreatic intraepithelial neoplasia and intraductal pa- pillary mucinous neoplasms. In: American Journal of Surgical Pathology 28, S. 977-987.
- Hu YX, Watanabe H, Li P et al. (2000): An immunohistochemical analysis of p27 expression in human pancreatic carcinomas. In: Pancreas 21, S. 226-230.
- Hutvágner G, Simard MJ (2008): Argonaute proteins: key players in RNA si- lencing. In: Nature 9, S. 22-32.
- Nykänen A, Haley B, Zamore P (2001): ATP requirements and small interfer- ing RNA structure in the RNA Interference pathway. In: Cell 107, S. 309- 321.
- Leitlinienprogramm Onkologie (Deutsche Krebsgesellschaft, Deutsche Krebshilfe, AWMF): S3-Leitlinie Exokrines Pankreaskarzinom, Kurzversion 1.0, 2013 AWMF Registernummer:032-010OL, http://leitlinienprogramm- onkologie.de/Leitlinien.7.0.html
- Beger HG, Rau B, Gansauge F et al. (2008): Bauchspeicheldrüsenkrebs - Heilungschancen minimal. In: Deutsches Ärzteblatt 105, S. 255-262.
- McGinnis S, Madden TL (2004): BLAST: at the core of a powerful and di- verse set of sequence analysis tools. In: Nucleic Acids Research 32, S. W20- W25.
- Bardeesy N, Aguirre AJ, Chu GC et al. (2006): Both p16 Ink4a and the p19 Arf - p53 pathway constrain progression of pancreatic adenocarcinoma in the mouse. In: Proceedings of the National Academy of Sciences 103, S. 5947- 5952.
- Popov KM, Zhao Y, Shimomura Y et al. (1992): Branched-chain a-Ketoacid Dehydrogenase Kinase: Molecular cloning, expression, and sequence similar- ity with histidine protein kinases. In: Journal of Biological Chemistry 267, S. 13127-13130.
- Shimomura Y, Honda T, Shiraki M et al. (2006): Branched-chain amino acid catabolism in exercise and liver disease. In: Journal of Nutrition 136, S. 250S-253S.
- Sweatt AJ, Wood M, Suryawan A et al. (2004): Branched-chain amino acid catabolism: unique segregation of pathway enzymes in organ systems and pe- ripheral nerves. In: American Journal of Physiology: Endocrinology and Me- tabolism 286, S. E64-E76.
- Goggins M, Hruban RH, Kern SE (2000): BRCA2 is inactivated late in the development of pancreatic intraepithelial neoplasia. In: American Journal of Pathology 156, S. 1767-1771.
- Hwang RF, Moore T, Arumugam T et al. (2008): Cancer-associated stromal fibroblasts promote pancreatic tumor progression. In: Cancer Research 68, S. 918-926.
- Easton D, Thompson D, McGuffog L et al. (1999): Cancer risks in BRCA2 mutation carriers. In: Journal of the National Cancer Institute 91, S. 1310- 1316.
- Cohen GM (1997): Caspases: the executioners of apoptosis. In: Biochemical Journal 326, S. 1-16.
- Schriever SC, Deutsch MJ, Adamski J et al. (2013): Cellular signaling of amino acids towards mTOR1 activation in impaired human leucine catabo- lism. In: Journal of Nutritional Biochemistry 24, S. 824-831.
- Ota T, Suzuki Y, Nishikawa T et al. (2004): Complete sequencing and char- acterization of 21,243 full-length human cDNAs. In: Nature Genetics 36, S. 40-45.
- Lewis BP, Burge CB, Bartel DP (2005): Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are mi- croRNA targets. In: Cell 120, S. 15-20.
- Lukas J, Bartkova J, Bartek J (1996): Convergence of mitogenic signalling cascades from diverse classes of receptors at the Cyclin D-Cyclin-dependent kinase-pRb-controlled G1 checkpoint. In: Molecular and Cellular Biology 16, S. 6917-6925.
- Pagano M, Pepperkok R, Verde F et al. (1992): Cyclin A is required at two points in the human cell cycle. In: The EMBO Journal 11, S. 961-971.
- Komar G, Kauhanen S, Liukko K et al. (2009): Decreased blood flow with increased metabolic activity: a novel sign of pancreatic tumor aggressiveness. In: Clinical Cancer Research 15, S. 5511-5517.
- Popov KM, Zhao Y, Shimomura Y et al. (1995): Dietary control and tissue specific expression of Branched-Chain α-Ketoacid Dehydrogenase Kinase. In: Archives of Biochemistry and Biophysics 316, S. 148-154.
- Huang YS, Chuang DT (1999): Down-regulation of rat mitochondrial Branched-Chain 2-Oxoacid Dehydrogenase Kinase gene expression by glu- cocorticoids. In: Biochemical Journal 339, S. 503-510.
- Parsa I, Longnecker DS, Scarpelli DG et al. (1985): Ductal metaplasia of hu- man exocrine pancreas and its association with carcinoma. In: Cancer Re- search 45, S. 1285-1290.
- Lüttges J, Reinecke-Luthge A, Mollmann B et al. (1999): Duct changes and K-ras mutations in the disease-free pancreas: analysis of type, age relation and spatial distribution. In: Virchows Archiv 435, S. 461-468.
- May ME, Buse MG (1989): Effects of branched-chain amino acids on protein turnover. In: Diabetes/ Metabolism Research and Reviews 5, S. 227-245.
- Hruban RH, Maitra A, Schulick R et al. (2008): Emerging molecular biology of pancreatic cancer. In: Gastrointestinal Cancer Research 2, S. 10-15.
- Blaine SA, Ray KC, Branch KM et al. (2009): Epidermal growth factor re- ceptor regulates pancreatic fibrosis. In: American Journal of Physiology - Gastrointestinal and Liver Physiology 297, S. 434-441.
- Kobayashi R, Shimomura Y, Otsuka M et al. (2000): Experimental hyperthy- roidism causes inactivation of the Branched-Chain alpha-Ketoacid Dehydro- genase Complex in rat liver. In: Archives of Biochemistry and Biophysics 375, S. 55-61.
- Jackson AL, Bartz SR, Schelter J et al. (2003): Expression profiling reveals off-target gene regulation by RNAi. In: Nature Biotechnology 21, S. 635- 638.
- Jaffee EM, Hruban RH, Canto M et al. (2002): Focus on pancreas cancer. In: Cancer Cell 2, S. 25-28.
- Elbashir SM, Martinez J, Patkaniowska A et al. (2001): Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. In: The EMBO Journal 20, S. 6877-6888.
- Yamano M, Fujii H, Takagaki T et al. (2000): Genetic progression and diver- gence in pancreatic carcinoma. In: American Journal of Pathology 156, S. 2123-2133.
- Chi JT, Chang HY, Wang NN et al. (2003): Genomewide view of gene si- lencing by small interfering RNAs. In: Proceedings of the National Academy of Sciences 100, S. 6343-6346.
- Ui-Tei K, Naito Y, Takahashi F et al. (2004): Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interfer- ence. In: Nucleic Acids Research 32, S. 936-948.
- Thayer SP, Di Magliano MP, Heiser PW (2003): Hedgehog is an early and late mediator of pancreatic cancer tumorigenesis. In: Nature 425, S. 851-856.
- Cattaneo M, Canton C, Albertini A et al. (2004): Identification of a region within SEL1L protein required for tumour growth inhibition. In: Gene 326, S. 149-156.
- Li C, Heidt DG, Dalerba P et al. (2007): Identification of pancreatic stem cells. In: Cancer Research 67, S. 1030-1037.
- Joshi MA, Jeoung NH, Obayashi M et al. (2006): Impaired growth and neuro- logical abnormalities in Branched-Chain α-Keto acid Dehydrogenase Kinase- deficient mice. In: Biochemical Journal 400, S. 153-162.
- Oliver FJ, De la Rubia G, Rolli V et al. (1998): Importance of Poly (ADP- ribose) Polymerase and its cleavage in apoptosis. In: The Journal of Biologi- cal Chemistry 273, S. 33533-33539.
- Burris HA 3rd, Moore MJ, Andersen J et al. (1997): Improvements in surviv- al and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: a randomized trial. In: Journal of Clinical Oncolo- gy 15, S. 2403-2413.
- Wilentz RE, Geradts J, Maynard R et al. (1998): Inactivation of the p16 (INK4A) tumor-suppressor gene in pancreatic duct lesions: loss of intranu- clear expression. In: Cancer Research 58, S. 4740-4744.
- Von Hoff DD, Ervin T, Arena FP et al. (2013): Increased survival in pancre- atic cancer with nab-Paclitaxel plus Gemcitabine. In: New England Journal of Medicine 369, S. 1691-1703.
- Hu YX, Watanabe H, Ohtsubo K et al. (1998): Infrequent expression of p21 is related to altered p53 protein in pancreatic carcinoma. In: Clinical Cancer Research 4, S. 1147-1152.
- Olive KP, Jacobetz MA, Davidson CJ et al. (2009): Inhibition of Hedgehog signaling enhances delivery of chemotherapy in a mouse model of pancreatic cancer. In: Science 324, S. 1457-1461.
- Bartlett DW, Davis MW (2006): Insights into the kinetics of siRNA-mediated gene silencing from live-cell and live-animal bioluminescent imaging. In: Nucleic Acids Research 34, S. 322-333.
- Ott C, Heinmöller E, Gaumann A et al. (2007): Intraepitheliale Neoplasien (PanIN) und intraduktale papillär-muzinöse Neoplasien (IPMN) des Pankreas als Vorläufer des Pankreaskarzinoms. In: Medizinische Klinik -Intensivme- dizin und Notfallmedizin 102, S. 127-135.
- Mattick JS (1994): Introns: evolution and function. In: Current Opinion in Genetics and Development 4, S. 823-831.
- Chen J, Zhang W (2012): Kinetic analysis of the effects of target structure on siRNA efficiency. In: The Journal of Chemical Physics 137:225102.
- Terhune PG, Phifer DM, Tosteson TD et al. (1998): K-ras mutations in focal proliferative lesions of human pancreas. In: Cancer Epidemiology Bi- omarkers and Prevention 7, S. 515-521.
- Wilentz RE, Iacobuzio-Donahue CA, Argani P et al. (2000): Loss of expres- sion of Dpc4 in pancreatic intraepithelial neoplasia: evidence that DPC4 inac- tivation occurs late in neoplastic progression. In: Cancer Research 60, S. 2002-2006.
- Peinemann F, Danner DJ (1994): Maple syrup urine disease 1954 to 1993. In: Journal of Inherited Metabolic Disease 17, S. 3-15.
- Harris RA, Joshi M, Jeoung NH (2004): Mechanisms responsible for regula- tion of branched-chain amino acid catabolism. In: Biochemical and Biophysi- cal Research Communications 313, S. 391-396.
- Popov KM, Hawes JW, Harris RA (1997): Mitochondrial alpha-ketoacid de- hydrogenase kinases: a new family of protein kinases. In: Advances in Sec- ond Messenger Phosphoprotein Research 31, S. 105-111.
- Oyarzabal A, Bravo-Alonso I, Sánchez-Aragó M et al. (2016): Mitochondrial response to the BCKDK-deficiency: Some clues to understand the positive dietary response in this form of autism. In: Biochimica et Biophysica Acta 1862, S. 592-600.
- Buchholz M, Gress TM (2009): Molecular changes in pancreatic cancer. In: Expert Review of Anticancer Therapy 9, S. 1487-1497.
- Harris RA, Popov KM, Kedishvili NY et al. (1993): Molecular cloning of the Branched-Chain alpha-Keto acid Dehydrogenase Kinase and the CoA- dependent Methylmalonate Semialdehyde Dehydrogenase. In: Advances in Enzyme Regulation 33, S. 255-265.
- Wynn RM, Kato M, Machius M et al. (2004): Molecular mechanism for regu- lation of the human mitochondrial Branched-Chain α-Ketoacid Dehydrogen- ase Complex by phosphorylation. In: Structure 12, S. 2185-2196.
- Cubilla AL, Fitzgerald PJ (1976): Morphological lesions associated with hu- man primary invasive nonendocrine pancreatic cancer. In: Cancer Research 36, S. 2690-2698.
- Cubilla AL, Fitzgerald PJ (1975): Morphological patterns of primary nonen- docrine human pancreas carcinoma. In: Cancer Research 35, S. 2234-2248.
- Zhen H, Kitaura Y, Kadota Y et al. (2016): mTORC1 is involved in the regu- lation of branched-chain amino acid catabolism in mouse heart. In: Federation of European Biochemical Societies Open Bio 6, S. 43-49.
- Morton JP, Timpson P, Karim SA et al. (2010): Mutant p53 drives metastasis and overcomes growth arrest/ senescence in pancreatic cancer. In: Proceed- ings of the National Academy of Sciences 107, S. 246-251.
- Novarino G, El-Fishawy P, Kayserili H et al. (2012): Mutations in BCKD- Kinase lead to a potentially treatable form of autism with epilepsy. In: Sci- ence 338, S. 394-397.
- Redston MS, Caldas C, Seymour AB et al. (1994): p53 mutations in pancreat- ic carcinoma and evidence of common involvement of homocopolymer tracts in DNA microdeletions. In: Cancer Research 54, S. 3025-3033.
- Harris RA, Popov KM, Zhao Y (1995): Nutritional regulation of the protein kinases responsible for the phosphorylation of the a-Ketoacid Dehydrogenase Complexes. In: Journal of Nutrition 125, S. 1758S-1761S.
- Oettle H, Heinemann V, Herrmann R et al. (2010): Onkopedia Leitlinie Pan- kreaskarzinom. In: Deutsche Gesellschaft für Hämatologie und Medizinische Onkologie e.V., https://www.onkopedia.com/de/wissensdatenbank/wissens- datenbank/pankreaskarzinom/AWMFS3Leitlinie2013.pdf.
- Biankin AV, Kench JG, Morey AL et al. (2001): Overexpression of p21 WAF1/CIP1 is an early event in the development of pancreatic intraepithelial neoplasia. In: Cancer Research 61, S. 8830-8837.
- Tsutsumi S, Hogan V, Nabi IR et al. (2003): Overexpression of the Autocrine Motility Factor/ Phosphoglucose Isomerase induces transformation and sur- vival of NIH-3T3 fibroblasts. In: Cancer Research 63, S. 242-249.
- Harris RA, Joshi M, Jeoung NH et al. (2005): Overview of the molecular and biochemical basis of branched-chain amino acid catabolism. In: Journal of Nutrition 135, S. 1527S-1530S.
- Xiong Y, Hannon GJ, Zhang H et al. (1993): p21 is a universal inhibitor of cyclin kinases. In: Nature 366, S. 701-704.
- Kuwano K, Kunitake R, Kawasaki M et al. (1996): P21Waf1/Cip1/Sdi1 and p53 expression in association with DNA strand breaks in idiopathic pulmo- nary fibrosis. In: American Journal of Respiratory and Critical Care Medicine 154, S. 477-483.
- Toyoshima H, Hunter T (1994): p27, a novel inhibitor of G1 cyclin-Cdk pro- tein kinase activity, is related to p21. In: Cell 78, S. 67-74.
- Korc M (2007): Pancreatic cancer-associated stroma production. In: Ameri- can Journal of Surgery 194, S. 84-86.
- Real FX, Cibrián-Uhalte E, Martinelli P (2008): Pancreatic cancer develop- ment and progression: remodeling the model. In: Gastroenterology 135, S. 724-728.
- Petersen GM, De Andrade M, Goggins M et al. (2006): Pancreatic cancer ge- netic epidemiology (PACGENE) consortium. In: Cancer Epidemiology Bi- omarkers and Prevention 15, S. 704-710.
- Friess H, Berberat P, Schilling M et al. (1996): Pancreatic cancer: the poten- tial clinical relevance of alterations in growth factors and their receptors. In: Journal of Molecular Medicine 74, S. 35-42.
- Hruban RH, Adsay NV, Albores-Saavedra J et al. (2001): Pancreatic intraepi- thelial neoplasia: a new nomenclature and classification system for pancreatic duct lesions. In: American Journal of Surgical Pathology 25, S. 579-586.
- Zhang L, Sanderson SO, Lloyd RV et al. (2007): Pancreatic intraepithelial neoplasia in heterotopic pancreas: evidence for the progression model of pan- creatic ductal adenocarcinoma. In: American Journal of Surgical Pathology 31, S. 1191-1195.
- Vonlaufen A, Joshi S, Qu C et al. (2008): Pancreatic stellate cells: partners in crime with pancreatic cancer cells. In: Cancer Research 68, S. 2085-2093.
- Apte MV, Haber PS, Applegate TL et al. (1998): Periacinar stellate shaped cells in rat pancreas: identification, isolation, and culture. In: Gut 43, S. 128- 133.
- Lynch CJ, Halle B, Fujii H et al. (2003): Potential role of leucine metabolism in the leucine-signaling pathway involving mTOR. In: American Journal of Physiology: Endocrinology and Metabolism 285, S. E854-E863.
- Hruban RH, Goggins M, Parsons J et al. (2000): Progression model for pan- creatic cancer. In: Clinical Cancer Research 6, S. 2969-2972.
- Harris RA, Popov KM, Shimomura Y et al. (1992): Purification, characteriza- tion, regulation and molecular cloning of mitochondrial protein kinases. In: Advances in Enzyme Regulation 32, S. 267-284.
- Odessey R (1982): Purification of rat kidney Branched-Chain Oxo acid De- hydrogenase Complex with endogenous kinase activity. In: Biochemical Journal 204, S. 353-356.
- Cook KG, Bradford AP, Yeaman SJ et al. (1984): Regulation of bovine kid- ney Branched-Chain 2-Oxoacid Dehydrogenase Complex by reversible phos- phorylation. In: European Journal of Biochemistry 145, S. 587-591.
- Shimomura Y, Obayashi M, Murakami T et al. (2001): Regulation of branched-chain amino acid catabolism: nutritional and hormonal regulation of activity and expression of the Branched-Chain alpha-Keto acid Dehydrogen- ase Kinase. In: Current Opinion in Clinical Nutrition and Metabolic Care 4, S. 419-423.
- Harris RA, Kobayashi R, Murakami T et al. (2001): Regulation of Branched- Chain α-Keto acid Dehydrogenase Kinase expression in rat liver. In: Journal of Nutrition 131, S. 841S-845S.
- Doisaki M, Katano Y, Nakano I et al. (2010): Regulation of hepatic Branched-Chain alpha-Keto acid Dehydrogenase Kinase in a rat model for type 2 diabetes mellitus at different stages of the disease. In: Biochemical and Biophysical Research Communications 393, S. 303-307.
- Obayashi M, Sato Y, Harris RA et al. (2001): Regulation of the activity of Branched-Chain 2-Oxo acid Dehydrogenase (BCODH) Complex by binding BCODH kinase. In: Federation of European Biochemical Societies Letters 491, S. 50-54.
- Harris RA, Zhang B, Goodwin GW et al. (1990): Regulation of the Branched- Chain alpha-Ketoacid Dehydrogenase and elucidation of a molecular basis for maple syrup urine disease. In: Advances in Enzyme Regulation 30, S. 245- 263.
- Dinger ME, Mercer TR, Mattick JS (2008): RNAs as extracellular signaling molecules. In: Journal of Molecular Endocrinology 40, S. 151-159.
- Ellenrieder V, Alber B, Lacher U et al. (2000): Role of MT-MMPs and MMP-2 in pancreatic cancer progression. In: International Journal of Cancer 85, S. 14-20.
- Liggett WH Jr, Sidransky D (1998): Role of the p16 tumor suppressor gene in cancer. In: Journal of Clinical Oncology 16, S. 1197-1206.
- Zhao Y, Samal E, Srivastava D (2005): Serum response factor regulates a muscle-specific microRNA that targets Hand2 during cardiogenesis. In: Na- ture 436, S. 214-220.
- Scacheri PC, Rozenblatt-Rosen O, Caplen NJ et al. (2004): Short interfering RNAs can induce unexpected and divergent changes in the levels of untarget- ed proteins in mammalian cells. In: Proceedings of the National Academy of Sciences 101, S. 1892-1897.
- Saxena S, Jónsson ZO, Dutta A (2003): Small RNAs with imperfect match to endogenous mRNA repress translation. In: The Journal of Biological Chemis- try 278, S. 44312-44319.
- Bailey JM, Swanson BJ, Hamada T (2008): Sonic hedgehog promotes desmoplasia in pancreatic cancer. In: Clinical Cancer Research 14, S. 5995- 6004.
- Neesse A, Michl P, Frese KK et al. (2011): Stromal biology and therapy in pancreatic cancer. In: Gut 60, S. 861-868.
- Neesse A, Algül H, Tuveson DA et al. (2015): Stromal biology and therapy in pancreatic cancer: a changing paradigm. In: Gut 64, S.1476-1484.
- Chu GC, Kimmelman AC, Hezel AF et al. (2007): Stromal biology of pan- creatic cancer. In: Journal of Cellular Biochemistry 101, S. 887-907.
- Rhim AD, Oberstein PE, Thomas DH et al. (2014): Stromal elements act to restrain, rather than support, pancreatic ductal adenocarcinoma. In: Cancer Cell 25, S. 735-747.
- Tso SC, Qi X, Gui WJ et al. (2013): Structure-based design and mechanisms of allosteric inhibitors for mitochondrial Branched-Chain α-Ketoacid Dehy- drogenase Kinase. In: Proceedings of the National Academy of Sciences 110, S. 9728-9733.
- Machius M, Chuang JL, Wynn RM et al. (2001): Structure of rat BCKD ki- nase: Nucleotide-induced domain communication in a mitochondrial protein kinase. In: Proceedings of the National Academy of Sciences 98, S. 11218- 11223.
- Harris RA, Hawes JW, Popov KM et al. (1997): Studies on the regulation of the mitochondrial α-Ketoacid Dehydrogenase Complexes and their kinases. In: Advances in Enzyme Regulation 31, S. 271-293.
- Hackeng WM, Hruban RH, Offerhaus GJA et al. (2016): Surgical and molec- ular pathology of pancreatic neoplasms. In: Diagnostic Pathology 11, S. 47- 64.
- Sudbery I, Enright AJ, Fraser AG et al. (2010): Systematic analysis of off- target effects in an RNAi screen reveals microRNAs affecting sensitivity to TRAIL-induced apoptosis. In: BMC Genomics 11:175.
- Suehara N, Mizumoto K, Muta T et al. (1997): Telomerase elevation in pan- creatic ductal carcinoma compared to nonmalignant pathological states. In: Clinical Cancer Research 3, S. 993-998.
- Van Heek NT, Meeker AK, Kern SE et al. (2002): Telomere shortening is nearly universal in pancreatic intraepithelial neoplasia. In: American Journal of Pathology 161, S. 1541-1547.
- Hutson SM (2006): The case for regulating indispensable amino acid metabo- lism: the Branched-Chain α-Keto acid Dehydrogenase Kinase-knockout mouse. In: Biochemical Journal 400, S. E1-E3.
- Nazli O, Bozdag AD, Tansug T et al. (2000): The diagnostic importance of CEA and CA 19-9 for the early diagnosis of pancreatic carcinoma. In: Hepa- togastroenterology 47, S. 1750-1752.
- Manning G, Whyte DB, Martinez R et al. (2002): The protein kinase com- plement of the human genome. In: Science 298, 1912-1934.
- Ui-Tei K, Naito Y, Nishi K et al. (2008): Thermodynamic stability and Wat- son-Crick base pairing in the seed duplex are major determinants of the effi- ciency of the siRNA-based off-target effect. In: Nucleic Acids Research 36, S. 7100-7109.
- Funasaka T, Raz A (2007): The role of Autocrine Motility Factor in tumor and tumor microenvironment. In: Cancer and Metastasis Reviews 26, S. 725- 735.
- Kamola PJ, Nakano Y, Takahashi T et al. (2015): The siRNA non-seed re- gion and its target sequences are auxiliary determinants of off-target effects. In: Public Library of Science Computational Biology 11(12): e1004656.
- Gerhard DS (2004): The status, quality, and expansion of the NIH full-length cDNA project: The Mammalian Gene Collection (MGC). In: Genome Re- search 14, S. 2121-2127.
- Malaisse WJ, Hutton JC, Carpinelli AR et al. (1980): The stimulus-secretion coupling of amino acid-induced insulin release: metabolism and cationic ef- fects of leucine. In: Diabetes 29, S. 431-437.
- Muller EA, Danner DJ (2004): Tissue-specific translation of murine Branched-chain α-Ketoacid Dehydrogenase Kinase mRNA is dependent upon an upstream open reading frame in the 5'-untranslated region. In: Journal of Biological Chemistry 279, S. 44645-44655.
- Poot M (2013): Towards identification of individual etiologies by resolving genomic and biological conundrums in patients with autism spectrum disor- ders. In: Molecular Syndromology 4, S. 213-226.
- Buchholz M, Braun M, Heidenblut A et al. (2005): Transcriptome analysis of microdissected pancreatic intraepithelial neoplastic lesions. In: Oncogene 24, S. 6626-6636.
- Morton JP, Klimstra DS, Mongeau ME et al. (2008): Trp53 deletion stimu- lates the formation of metastatic pancreatic tumors. In: The American Journal of Pathology 172, S. 1081-1087.
- Liotta LA, Mandler R, Murano G et al. (1986): Tumor cell autocrine motility factor. In: Proceedings of the National Academy of Sciences USA 83, S. 3302-3306.
- Watanabe H, Takehana K, Date M et al. (1996): Tumor cell Autocrine Motili- ty Factor is the Neuroleukin/ Phosphohexose Isomerase Polypeptide 1. In: Cancer Research 56, S. 2960-2963.
- Mahadevan D, Von Hoff DD (2007): Tumor-stroma interactions in pancreatic ductal adenocarcinoma. In: Molecular Cancer Therapy 6, S. 1186-1197.
- García-Cazorla A, Oyarzabal A, Fort J et al. (2014): Two novel mutations in the BCKDK (Branched-Chain Keto-acid Dehydrogenase Kinase) gene are re- sponsible for a neurobehavioral deficit in two pediatric unrelated patients. In: Human Mutation 35, S. 470-477.