Endometriosis and the expression of invasion factors in both the endometrium and the peritoneum

Endometriose und die Expression von Invasionsfaktoren im Endometrium und im Peritoneum Hintergrund: Endometriose ist eine \uf6strogen-abh\ue4ngige Entz\ufcndungserkrankung, die bis zu 10% der geb\ue4rf\ue4higen Frauen betreffen kann. Verschiedene Invasionsfaktoren einschlie flich Matrix Metalloproteinasen (MMPs) und ADAMs (A Disintegrin and Metalloproteinase) f\ufchren zu einem fr\ufchen Defekt in der physiologischen Aktivit\ue4t des Endometriums und die darauffolgende Wucherung aus der Geb\ue4rmutterh\uf6hle. Sogenannte Gewebeinhibitoren von Metalloproteinasen (TIMPs) sind die funktionellen Inhibitoren von MMPs und dadurch kann ein MMP-TIMP Ungleichgewicht zur Entwicklung ektoper L\ue4sionen beitragen. In dieser Studie wurden insgesamt 114 pr\ue4menopausale Frauen mit oder ohne Unfruchtbarkeit, die sich einer laparoskopischen Chirurgie auf Verdacht einer Endometriose unterzogen haben, einbezogen. Methoden: MMP-9, MMP-28, TIMP-1, ADAM-10 und ADAM-17 mRNA Expressionen von 53 Endometriose-freien Patienten und 61 Endometriose Patienten wurden mittels Real-Time quantitative PCR (qRT-PCR) analysiert. Insgesamt wurden 151 frische endometriale Gewebsproben, die 62 ektope beinhalten und 54 peritoneale Gewebsproben untersucht. Ergebnisse: Im Vergleich zum eutopen Endometrium waren die Expressionswerte von MMP-9, MMP-28, ADAM-10 und ADAM-17 im ektopen Endometrium signifikant erh\uf6ht. In gepaarten F\ue4llen war die TIMP-1 Expression im ektopen Endometrium signifikant niedriger als im eutopen Endometrium. Schlussfolgerung: Diese Resultate lassen darauf schlie fen, dass MMP-9, MMP-28, TIMP-1, ADAM-10 und ADAM-17 an der invasiven Erkrankung der Endometriose beteiligt sind. Des Weiteren zeigen die Ergebnisse, dass jenes eutope Endometrium von Endometriose-Patienten anf\ue4lliger f\ufcr peritoneale Implantationen zu sein scheint als das von Kontrollpatienten. Schl\ufcssel W\uf6rter: Endometriose, endometriale Invasionsfaktoren, Peritoneum, MMP-9, MMP-28, TIMP-1, ADAM-10, ADAM-17Endometriosis and the Expression of Invasion Factors both in the Endometrium and the Peritoneum Background: Endometriosis is a common oestrogen-dependent inflammatory disease which affects up to 10% of women during their reproductive years. Different invasion factors including Matrix Metalloproteinases (MMPs) and ADAMs (A Disintegrin and Metalloproteinase) lead to an early defect in the physiological activity of the endometrium, which result in its overgrowth outside the uterine cavity. Tissue inhibitors of metalloproteinases (TIMPs) are the functional inhibitors of MMPs and therefore MMP-TIMP imbalance may contribute to the development of ectopic lesions. A total of 114 premenopausal women were recruited in this study who underwent laparoscopic surgery due to the suspicion of endometriosis with or without infertility. Methods: MMP-9, MMP-28, TIMP-1, ADAM-10 and ADAM-17 mRNA expressions were analyzed from 53 controls and 61 patients with endometriosis by quantitative Real-Time PCR (qRT-PCR). Results: Significant higher MMP-9, MMP-28, ADAM-10 and ADAM-17 expressions were observed in the ectopic endometrium compared to the eutopic endometrium of controls and women with endometriosis. In paired cases TIMP-1 expression was significantly decreased in ectopic lesions compared to the eutopic endometrium of patients. Conclusion: These findings suggest that MMP-9, MMP-28, ADAM-10 and ADAM-17 contribute to invasive disease of endometriosis. Furthermore results show that the eutopic endometrium from endometriosis patients appears to be more invasive and prone to peritoneal implantation than that from women without the disease. Key Words: endometriosis, endometriotic invasion factors, peritoneum, MMP-9, MMP-28, TIMP-1, ADAM-10, ADAM-17submitted by Susanne GamperlWien, Univ. f\ufcr Bodenkultur, Dipl.-Arb., 2015Zsfassung in dt. Sprach

Endometriosis and the expression of invasion factors in both the endometrium and the peritoneum

Endometriose und die Expression von Invasionsfaktoren im Endometrium und im Peritoneum Hintergrund: Endometriose ist eine östrogen-abhängige Entzündungserkrankung, die bis zu 10% der gebärfähigen Frauen betreffen kann. Verschiedene Invasionsfaktoren einschließlich Matrix Metalloproteinasen (MMPs) und ADAMs (A Disintegrin and Metalloproteinase) führen zu einem frühen Defekt in der physiologischen Aktivität des Endometriums und die darauffolgende Wucherung aus der Gebärmutterhöhle. Sogenannte Gewebeinhibitoren von Metalloproteinasen (TIMPs) sind die funktionellen Inhibitoren von MMPs und dadurch kann ein MMP-TIMP Ungleichgewicht zur Entwicklung ektoper Läsionen beitragen. In dieser Studie wurden insgesamt 114 prämenopausale Frauen mit oder ohne Unfruchtbarkeit, die sich einer laparoskopischen Chirurgie auf Verdacht einer Endometriose unterzogen haben, einbezogen. Methoden: MMP-9, MMP-28, TIMP-1, ADAM-10 und ADAM-17 mRNA Expressionen von 53 Endometriose-freien Patienten und 61 Endometriose Patienten wurden mittels Real-Time quantitative PCR (qRT-PCR) analysiert. Insgesamt wurden 151 frische endometriale Gewebsproben, die 62 ektope beinhalten und 54 peritoneale Gewebsproben untersucht. Ergebnisse: Im Vergleich zum eutopen Endometrium waren die Expressionswerte von MMP-9, MMP-28, ADAM-10 und ADAM-17 im ektopen Endometrium signifikant erhöht. In gepaarten Fällen war die TIMP-1 Expression im ektopen Endometrium signifikant niedriger als im eutopen Endometrium. Schlussfolgerung: Diese Resultate lassen darauf schließen, dass MMP-9, MMP-28, TIMP-1, ADAM-10 und ADAM-17 an der invasiven Erkrankung der Endometriose beteiligt sind. Des Weiteren zeigen die Ergebnisse, dass jenes eutope Endometrium von Endometriose-Patienten anfälliger für peritoneale Implantationen zu sein scheint als das von Kontrollpatienten. Schlüssel Wörter: Endometriose, endometriale Invasionsfaktoren, Peritoneum, MMP-9, MMP-28, TIMP-1, ADAM-10, ADAM-17Endometriosis and the Expression of Invasion Factors both in the Endometrium and the Peritoneum Background: Endometriosis is a common oestrogen-dependent inflammatory disease which affects up to 10% of women during their reproductive years. Different invasion factors including Matrix Metalloproteinases (MMPs) and ADAMs (A Disintegrin and Metalloproteinase) lead to an early defect in the physiological activity of the endometrium, which result in its overgrowth outside the uterine cavity. Tissue inhibitors of metalloproteinases (TIMPs) are the functional inhibitors of MMPs and therefore MMP-TIMP imbalance may contribute to the development of ectopic lesions. A total of 114 premenopausal women were recruited in this study who underwent laparoscopic surgery due to the suspicion of endometriosis with or without infertility. Methods: MMP-9, MMP-28, TIMP-1, ADAM-10 and ADAM-17 mRNA expressions were analyzed from 53 controls and 61 patients with endometriosis by quantitative Real-Time PCR (qRT-PCR). Results: Significant higher MMP-9, MMP-28, ADAM-10 and ADAM-17 expressions were observed in the ectopic endometrium compared to the eutopic endometrium of controls and women with endometriosis. In paired cases TIMP-1 expression was significantly decreased in ectopic lesions compared to the eutopic endometrium of patients. Conclusion: These findings suggest that MMP-9, MMP-28, ADAM-10 and ADAM-17 contribute to invasive disease of endometriosis. Furthermore results show that the eutopic endometrium from endometriosis patients appears to be more invasive and prone to peritoneal implantation than that from women without the disease. Key Words: endometriosis, endometriotic invasion factors, peritoneum, MMP-9, MMP-28, TIMP-1, ADAM-10, ADAM-17submitted by Susanne GamperlWien, Univ. für Bodenkultur, Dipl.-Arb., 2015Zsfassung in dt. Sprach

Effekte von Tyrosinkinase-Inhibitoren (TKI) und Histaminrezeptor 1 (HR1) -Antagonisten auf neoplastische Mastzellen (MZ) des Hundes

Mastzelltumore (MZT), auch Mastozytome genannt, treten bei Hunden sehr häufig auf. Gutartige MZT weisen eine langsame Wachstumsrate und in der Regel eine gute Prognose auf. Im aggressiven MZT kommt es zu einem unkontrollierten Wachstum neoplastischer Mastzellen (MZ), die sich auch auf regionale Lymphknoten oder andere Organe ausbreiten können, was wiederum zu einer schlechten Prognose führen kann. Hunde mit MZT können unter bestimmten Symptomen wie Juckreiz, Magengeschwüre oder anaphylaktische Reaktionen leiden. Diese Symptome werden durch endogene, aktive Substanzen wie Histamin verursacht. Histamin wird von MZ ausgeschüttet und ist bei zahlreichen allergischen Reaktionen beteiligt. In der klinischen Praxis werden Histamin-Rezeptor 1 (HR1) - und HR2-Antagonisten verwendet, um diese Symptome zu kontrollieren. Behandlungsstrategien für MZT des Grades I oder II umfassen chirurgische Eingriffe und gegebenenfalls postoperative Strahlentherapie. Therapieansätze von MZT des Grades III beinhalten Strahlentherapie und / oder Chemotherapie, die oft einen mäßigen Erfolg haben. Aktuell sind Masitinib und Toceranib, zwei Tyrosinkinase-Inhibitoren (TKI), die gegen die Rezeptor-Tyrosinkinase KIT gerichtet sind, für die Behandlung von inoperablen oder metastasierten MZT bei Hunden zugelassen. Das klinische Ansprechen ist jedoch nicht von langer Dauer und Rückfälle können auftreten. Daher sind die Identifizierung und Entwicklung neuer systemischer Antitumor-Medikamente, die das Wachstum und die Freisetzung von Mediatoren von neoplastischen MZ regulieren, erforderlich. Für aggressive Formen von MZ-Neoplasien im Menschen wird der TKI Midostaurin eingesetzt. Midostaurin blockiert effektiv das Wachstum und die Aktivierung neoplastischer MZ. Kürzlich veröffentlichte Studien haben gezeigt, dass der Bruton-Tyrosinkinase (BTK)-Hemmer Ibrutinib, der zur Therapie verschiedener humaner Krankheiten, wie chronischer lymphatischer Leukämie eingesetzt wird, die IgE-abhängige Freisetzung von Histamin bei MZ und Basophilen (BA) inhibiert. Das Ziel meiner Dissertation war es die Wirkung von Ibrutinib und anderen zielgerichteten Medikamenten auf neoplastischen MZ des Hundes zu untersuchen. Im zweiten Teil konzentrierte ich mich auf HR1-Antagonisten und deren Effekte auf neoplastische MZ des Hundes. In der ersten Studie konnte ich zeigen, dass Ibrutinib in zwei etablierten Hunde-MZ-Linien, C2 und NI-1, sowie in primären MZT Zellen das Wachstum blockiert. Zusätzlich, inhibiert Ibrutinib die IgE-abhängige Freisetzung von Histamin in NI-1 Zellen und in primären MZT Zellen. Da sich die aktuelle Forschung über MZ-Neoplasien auf Arzneimittelkombinationen zur Überwindung von Resistenzen konzentriert, habe ich potentielle kooperative Wirkungen zwischen mehreren TKI untersucht. In Kombinationsexperimenten konnte ich feststellen, dass "Ibrutinib und Midostaurin" synergistische wachstumshemmende Effekte in C2 Zellen aufweisen. In der zweiten Studie wollte ich herausfinden, ob HR1-Antagonisten neben den mediatorhemmenden Eigenschaften noch weitere Effekte auf MZ ausüben. Neben dem HR1-Antagonisten Loratadin habe ich Desloratadin, Rupatadin, Cyproheptadin, Dimetinden und Diphenhydramin an C2 und NI-1 getestet. Ich konnte zeigen, dass Desloratadin und Rupatadin eine stärkere inhibierende Wirkung auf das Wachstum und das Überleben von Hunde-MZ-Linien haben als Cyproheptadin, Dimetinden und Diphenhydramin. Außerdem reduzierten Loratadin, Desloratadin und Rupatadin die IgE-abhängige Freisetzung von Histamin in NI-1 Zellen. Des Weiteren konnte ich zeigen, dass die Kombination „Loratadin und Midostaurin“ in beiden getesteten MZ-Linien additive wachstumshemmende Wirkung aufweist. Zusammengefasst deutet meine Arbeit darauf hin, dass die Verabreichung von Ibrutinib einen potenziellen neuen therapeutischen Ansatz im MZT beim Hund darstellen könnte. Darüber hinaus zeigen meine Ergebnisse, dass Desloratadin oder Rupatadin Vorteile gegenüber anderen getesteten HR1-Antagonisten bei der Therapie von MZT aufweisen könnten. Ob die Anwendung von Ibrutinib oder bestimmten HR1-Antagonisten bei MZT-Patienten klinisch relevant ist, muss noch in weiteren Studien geklärt werden.Mast cell tumors (MCTs), also named mastocytoma, are very common skin neoplasms in dogs. Benign MCTs have a slow growth rate and usually a good prognosis, whereas aggressive MCTs are characterized by an uncontrolled growth of neoplastic mast cells (MCs) with the potential to spread to regional lymph nodes or other organs, resulting in a poor prognosis. Dogs with MCT may suffer from certain symptoms like pruritus, gastric ulceration or even anaphylaxis, which are caused by MC-derived biologically active substances, such as histamine. In clinical practice, histamine receptor 1 (HR1) and HR2 antagonists are used to control these symptoms. Treatment strategies of grade I or II MCTs involve surgical excision and sometimes post-surgical radiotherapy. Therapy approaches of grade III MCTs involve radiotherapy and/or chemotherapy, which have moderate effects. Currently available targeted drugs approved for treatment of unresectable or metastasized canine MCTs are masitinib and toceranib, two tyrosine kinase inhibitors (TKIs) directed against the receptor tyrosine kinase KIT. However, clinical responses are often temporary and relapses can occur. Therefore, identification and development of new systemic anti-tumor drugs controlling growth and mediator release in neoplastic MCs are needed. In aggressive forms of human MC neoplasms, the TKI midostaurin has been described to block growth and activation of neoplastic MCs effectively. Recently, the Bruton´s tyrosine kinase (BTK) inhibitor ibrutinib, which is used for therapy of several human diseases, such as chronic lymphocytic leukemia, was found to counteract IgE-dependent histamine release in MCs and basophils (BAs). In my thesis I was interested to test the effects of ibrutinib and other targeted drugs on canine neoplastic MCs. The second part focused on the anti-neoplastic effects of HR1 antagonists on canine neoplastic MCs. In the first study, I was able to demonstrate that the BTK inhibitor Ibrutinib counteracts growth of two established canine MC lines, C2 and NI-1, as well as in primary MCs obtained from MCT samples. In addition, ibrutinib was found to counteract IgE-dependent histamine release in NI-1 cells and primary MCT cells. Since current research on MC neoplasms focuses on drug combinations to overcome drug resistance, I examined possible cooperative interactions between several TKIs. In drug-combination experiments I found that `ibrutinib and midostaurin´ caused synergistic growth-inhibitory effects in C2 cells. In the second study, I wanted to know if HR1 antagonists exert additional effects apart from mediator-inhibiting properties. Beside the HR1 antagonist loratadine, I have also tested desloratadine, rupatadine, cyproheptadine, dimetindene and diphenhydramine on C2 and NI-1 cells. I found that desloratadine and rupatadine exert more potent effects on growth and survival in canine MC lines than cyproheptadine, dimetindene and diphenhydramine. In addition, loratadine, desloratadine and rupatadine were found to reduce IgE-dependent histamine release in NI-1 cells. The combination `loratadine+midostaurin´ was found to produce additive growth-inhibitory effects in both tested MC lines. Together, my thesis indicates that targeting BTK by ibrutinib may represent a potential new therapeutic approach in canine MCT. In addition, desloratadine or rupatadine may have some advantages over other tested HR1 antagonists in therapy of MCT, due to more potent effects on proliferation and histamine release in neoplastic MCs. Whether the application of ibrutinib or certain HR1 antagonists in MCT patients is clinically relevant, remains to be elucidated.Abweichender Titel laut Übersetzung der Verfasserin/des VerfassersMedizinische Universität Wien, Diss., 202

Enhanced epithelial to mesenchymal transition (EMT) and upregulated MYC in ectopic lesions contribute independently to endometriosis

Abstract Background Epithelial to mesenchymal transition (EMT) is a process in which epithelial cells lose polarity and cell-to-cell contacts and acquire the migratory and invasive abilities of mesenchymal cells. These abilities are thought to be prerequisites for the establishment of endometriotic lesions. A hallmark of EMT is the functional loss of E-cadherin (CDH1) expression in epithelial cells. TWIST1, a transcription factor that represses E-cadherin transcription, is among the EMT inducers. SNAIL, a zinc-finger transcription factor, and its close relative SLUG have similar properties to TWIST1 and are thus also EMT inducers. MYC, which is upregulated by estrogens in the uterus by an estrogen response cis-acting element (ERE) in its promoter, is associated with proliferation in endometriosis. The role of EMT and proliferation in the pathogenesis of endometriosis was evaluated by analyzing TWIST1, CDH1 and MYC expression. Methods CDH1, TWIST1, SNAIL and SLUG mRNA expression was analyzed by qRT-PCR from 47 controls and 74 patients with endometriosis. Approximately 42 ectopic and 62 eutopic endometrial tissues, of which 30 were matched samples, were collected during the same surgical procedure. We evaluated TWIST1 and MYC protein expression by immunohistochemistry (IHC) in the epithelial and stromal tissue of 69 eutopic and 90 ectopic endometrium samples, of which 49 matched samples were analyzed from the same patient. Concordant expression of TWIST1/SNAIL/SLUG and CDH1 but also of TWIST1 and MYC was analyzed. Results We found that TWIST1, SNAIL and SLUG are overexpressed (p < 0.001, p = 0.016 and p < 0.001) in endometriosis, while CDH1 expression was concordantly reduced in these samples (p < 0.001). Similar to TWIST1, the epithelial expression of MYC was also significantly enhanced in ectopic endometrium compared to eutopic tissues (p = 0.008). We found exclusive expression of either TWIST1 or MYC in the same samples (p = 0.003). Conclusions Epithelial TWIST1 is overexpressed in endometriosis and may contribute to the formation of endometriotic lesions by inducing epithelial to mesenchymal transition, as CDH1 was reduced in ectopic lesions. We found exclusive expression of either TWIST1 or MYC in the same samples, indicating that EMT and proliferation contribute independently of each other to the formation of endometriotic lesions

Enhanced epithelial to mesenchymal transition (EMT) and upregulated MYC in ectopic lesions contribute independently to endometriosis

Background Epithelial to mesenchymal transition (EMT) is a process in which epithelial cells lose polarity and cell-to-cell contacts and acquire the migratory and invasive abilities of mesenchymal cells. These abilities are thought to be prerequisites for the establishment of endometriotic lesions. A hallmark of EMT is the functional loss of E-cadherin (CDH1) expression in epithelial cells. TWIST1, a transcription factor that represses E-cadherin transcription, is among the EMT inducers. SNAIL, a zinc-finger transcription factor, and its close relative SLUG have similar properties to TWIST1 and are thus also EMT inducers. MYC, which is upregulated by estrogens in the uterus by an estrogen response cis-acting element (ERE) in its promoter, is associated with proliferation in endometriosis. The role of EMT and proliferation in the pathogenesis of endometriosis was evaluated by analyzing TWIST1, CDH1 and MYC expression. Methods CDH1, TWIST1, SNAIL and SLUG mRNA expression was analyzed by qRT-PCR from 47 controls and 74 patients with endometriosis. Approximately 42 ectopic and 62 eutopic endometrial tissues, of which 30 were matched samples, were collected during the same surgical procedure. We evaluated TWIST1 and MYC protein expression by immunohistochemistry (IHC) in the epithelial and stromal tissue of 69 eutopic and 90 ectopic endometrium samples, of which 49 matched samples were analyzed from the same patient. Concordant expression of TWIST1/SNAIL/SLUG and CDH1 but also of TWIST1 and MYC was analyzed. Results We found that TWIST1, SNAIL and SLUG are overexpressed (p < 0.001, p = 0.016 and p < 0.001) in endometriosis, while CDH1 expression was concordantly reduced in these samples (p < 0.001). Similar to TWIST1, the epithelial expression of MYC was also significantly enhanced in ectopic endometrium compared to eutopic tissues (p = 0.008). We found exclusive expression of either TWIST1 or MYC in the same samples (p = 0.003).(VLID)486705

Ponatinib Exerts Multiple Effects on Vascular Endothelial Cells: Possible Mechanisms and Explanations for the Adverse Vascular Events Seen in CML Patients Treated with Ponatinib

Abstract The potent BCR-ABL1-targeting tyrosine kinase inhibitor (TKI) ponatinib is used for the treatment of patients with drug-resistant chronic myeloid leukemia (CML). However, an increased risk of development of cardiovascular events has been described in CML patients treated with ponatinib. The etiology of these adverse events is currently unknown. In an attempt to discover mechanisms underlying ponatinib-induced adverse vascular events, we have evaluated the effects of ponatinib in vitro on human vascular endothelial cells and on contraction of explanted mice aortic rings. In addition, we examined the effects of ponatinib on angiogenesis in vivo in a mouse model of hind limb ischemia. Ponatinib dose-dependently induced apoptosis in human coronary artery endothelial cells (HCAEC) in a caspase assay (relative apoptosis vs. 1% DMSO: ponatinib 50 nM: 1.79±0.38, p&lt;0.001; ponatinib 100 nM: 2.13±0.42, p&lt;0.001) and this drug effect could partially be blocked by addition of insulin (ponatinib 100 nM + insulin 5 µg/ml: 1.70±0.18, p&lt;0.05). In addition, ponatinib was found to inhibit the proliferation of human umbilical vein endothelial cells (HUVEC) and the human microvascular endothelial cell line HMEC-1, with IC50 values ranging between 100 and 250 nM (p&lt;0.05) as determined by thymidine-incorporation assay. Using a phospho-receptor tyrosine kinase assay in HCAEC, we found that ponatinib also inhibits fetal bovine serum-induced phosphorylation of the VEGF receptor KDR as well as phosphorylation of MER and insulin receptors, which play a role in angiogenesis, vascular homeostasis, and vessel protection. We also found that ponatinib (1 µM, 4 hours) increases adhesion of HUVEC to a plastic-surface compared to DMSO control (Figure). Based on clinical observations of vasoconstriction in ponatinib-treated patients, we also applied ponatinib on aortic rings harvested from C57BL/6 mice. Ponatinib (100 nM, overnight) enhanced norepinephrine-induced vasoconstriction (log EC50: control -7.76±0.06 vs. ponatinib -7.96±0.05, p&lt;0.05, n=6) and inhibited acetylcholine-mediated vasodilatation (log IC50: control -7.45±0.05 vs. ponatinib -7.06±0.1, p&lt;0.001) as shown by myography. These drug effects were blocked by inhibition of nitric oxide (using nitric oxide synthase inhibitor L-NNA, 100 µM) or COX (by applying diclofenac, 3 mg/l), suggesting that ponatinib promotes the generation of vasoconstricting prostanoids. Ponatinib effects were also blocked by the calcium channel blocker nifedipine (1 µM). In C57BL/6 mice, ponatinib (5 mg/kg/day for 35 days) was found to inhibit blood flow recovery in a hind limb ischemia model as shown by Laser-Doppler perfusion imaging after femoral artery ligation. The blood perfusion ratios of the ischemic limb vs. non-ischemic limb at week 5 were: control group: 0.67±0.07 vs. ponatinib: 0.56±0.1; p&lt;0.05). Ponatinib-treated mice also developed toe and foot necrosis more frequently than control mice (necrosis score: control: 0.3 vs. ponatinib: 1.3). In summary, ponatinib affects endothelial cell growth and vasomotor function in-vitro as well as blood flow recovery in a mouse model. These findings might help explain the occurrence of vascular events in CML patients treated with ponatinib and may lead to development of therapeutic strategies for prevention and treatment of ponatinib-induced adverse events. Figure Figure. Disclosures Kirchmair: Ariad: Research Funding. Valent:Ariad: Honoraria, Research Funding; Amgen: Honoraria; Deciphera Pharmaceuticals: Research Funding; Novartis: Honoraria, Research Funding; Celgene: Honoraria, Research Funding. </jats:sec