Supplement 1/15

Proceedings of the Annual Meeting of the Austrian Society of Haematology and Medical Oncology Frühjahrstagung 2015 der Österreichischen Gesellschaft für Hämatologie und Medizinische Onkologie und der Arbeitsgemeinschaft hämato-onkologischer Pflegepersonen in Österreich (AHOP)

Salzburg, 23.–25. April 2015

Congress President: Prim. Univ.-Prof. Dr. Richard Greil

Secretary: DDr. Lukas Weiss Dr. Gabriel Rinnerthaler

Department of Internal Medicine III Salzburg Cancer Research Institute Paracelsus Private Medical University Salzburg

memo · Vol. 8 · Suppl 1/15 

Published online: 9 April 2015

© Springer-Verlag Wien

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Young Investigator Oral Presentations

A01 Clonal evolution in relapsed or refractory Diffuse large B cell lymphoma Thomas Melchardt1, Clemens Hufnagl1, Lukas Weiss1, Nadja Kopp2, David Weinstock2, Daniel Neureiter3, Wolfgang Tränkenschuh3, Richard Greil1, Oliver Weigert4, Alexander Egle1  epartment of Internal Medicine III, Paracelsus Private D Medical University, Salzburg, Austria 2 Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, USA 3 Institute of Pathology, Paracelsus Private Medical University, Salzburg, Austria 4 Department of Internal Medicine III, Ludwig-Maximilians University, Munich, Germany 1

Introduction:  Multiple mutations associated with the bio­ logy of diffuse large b-cell lymphoma (DLBCL) have been identified, but there is only little information available about the role of certain mutations and their mechanisms in clonal evolution during relapse. Material and Methods:  We identified 27 patients with available histologically confirmed relapsed or refractory DLBCL in a cohort of 346 consecutively treated patients. Primary formalin fixed paraffin embedded tumor sample, sample of refractory or relapsed disease and matched germ line were available for targeted next generation sequencing. A targeted exon capture and next-generation sequencing of all coding exons of 104 selected genes known to be frequently mutated in lymphoma were performed on a HiSeq 2500. Results:  Non-synonymous mutations were present in 74 of the 104 genes tested. Individual tumor samples showed between 0 and 29 non-synonymous mutations (median: 10). Less than six non-synonymous mutations in the primary tumor were associated with a better median OS than more mutations (28 versus 15 months p = 0.031). The “early divergent” and the “late divergent” patterns of evolution, previously described in a limited set of patients using VDJ rearrangements, were validated in our cohort using NGS data and showed no influence on the clinical outcome. We also observed that different genes win in clonal selection during treatment challenge compared to lymphoma development. Conclusions:  Targeted re-sequencing is a feasible and informative approach in FFPE-fixed DLBCL. Three major types of clonal evolution; large global change, subclonal selection and no or minimal change assuming preprogrammed resistance; were detected during relapse.

A02 An update of functional data regarding the pro-atherogenic and anti-angiogenic effects of nilotinib in patients with Ph+ CML

Susanne Herndlhofer1, Markus Theurl4, Sabine Cerny-Reiterer1,2, Gregor Hoermann7, Wolfgang R. Sperr1,2, Uwe Rix5, Irina Sadovnik1, Bernd Jilma8, Gerit-Holger Schernthaner9, Johann Wojta6, Dominik Wolf10, Giulio Superti-Furga5, Rudolf Kirchmair4, Peter Valent1,2  epartment of Internal Medicine I, Medical University of D Vienna, Vienna, Austria 2 Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria 3 Clinic for Small Animals, University of Veterinary Medicine Vienna, Vienna, Austria 4 Department of Internal Medicine I, Medical University of Innsbruck, Innsbruck, Austria 5 Research Center for Molecular Medicine (CeMM), Vienna, Austria 6 Department of Internal Medicine II, Clinical Division of Cardiology, Medical University of Vienna, Vienna, Austria 7 Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria 8 Department of Clinical Pharmacology, Medical University of Vienna, Vienna, Austria 9 Department of Internal Medicine II, Division of Angiology, Medical University of Vienna, Vienna, Austria 10 Department of Hematology & Oncology, University Hospital Bonn, Bonn, Germany 1

Introduction:  Nilotinib treatment of CML patients is associated with arterial occlusive disease (AOD). In this study, we performed additional experiments to further evaluate the underlying mechanisms. Materials and Methods:  We examined the pro-atherogenic and anti-angiogenic effects of imatinib and nilotinib on endothelial cells in in vitro studies as well as in vivo in patients and in two mouse models. Results:  We found a time-dependent accumulation of AOD in 36 nilotinib-treated patients (26.5% at 24 months vs. 44.4% at 44 months). In ApoE knock-out mice, treatment with nilotinib promoted atherosclerosis compared to control-mice (plaquearea: control: 16  ± 3.2 %; nilotinib: 22.4  ± 2.9). In a mouse model of hindlimb-ischemia, nilotinib-treated mice showed a decreased reperfusion and decreased microvessel-density compared to imatinib-treated mice (p < 0.05). In addition, we found that nilotinib promotes expression of the pro-atherogenic molecule VCAM-1 in C57BL/6 mice compared to imatinibtreated animals (cells/high-power-field: imatinib: 14.9 ± 11; nilotinib: 22.6 ± 9.3). We also examined the effects of nilotinib on human umbilical vein endothelial cells (HUVEC) and found that nilotinib, but not imatinib, promotes the expression of the cytoadhesion molecules ICAM-1, VCAM-1 and E-Selectin. In addition, nilotinib but not imatinib was found to inhibit the proliferation of HUVEC (IC50: 1 versus. 10 µM). Finally, we examined BM microvessel-density in CML patients before and one year after nilotinib-treatment and found that the numbers of endothelial cells per high-power-field decreases substantially (pre-nilotinib: 12 ± 2 versus post-nilotinib: 5 ± 2). Conclusions:  Nilotinib exerts direct pro-atherogenic and anti-angiogenic effects on endothelial cells. Whether these effects contribute to nilotinib-associated vasculopathy in patients with CML is currently under investigation.

Emir Hadzijusufovic1,2,3, Karin Albrecht-Schgoer4, Kilian Huber5, Florian Grebien5, Gregor Eisenwort1,2, Wilfried Schgoer4, Christoph Kaun6,

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A04

The TYK2-pathway as a novel therapeutic target in aggressive T-Cell Lymphoma

Evaluation of circulating cell-free DNA as a molecular monitoring tool during cancer therapy

Nicole Prutsch1, Takaomi Sanda6, Mathias Müller3, Ulrich Jäger4, Philipp Staber4, Bernd Boidol5, Suzanne Turner8, Lawren Wu7, Lukas Kenner1,2, Olaf Merkel1

Clemens Hufnagl1, Lukas Weiss1, Thomas Melchardt1, Martin Moik1, Daniela Asslaber1, Philipp Steininger4, Thomas Meißnitzer3, Daniel Neureiter2, Richard Greil1, Alexander Egle1

Institute of Pathology, Medical University Vienna, Vienna, Austria 2 Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria 3 Institute of Animal Breeding and Genetics, University of Veterinary Medicine, Vienna, Austria 4 Department of Medicine I, Division of Hematology and Hemostaseology and the Comprehensive Cancer Center Vienna, Vienna, Austria 5 Chemical Screening and Platform Austria for Chemical Biology, CeMM, Medical University of Vienna, Vienna, Austria 6 Cancer Science Institute of Singapore, National University of Singapore, Singapore, Singapore 7 Department of Immunology, Genentech Inc., South San Francisco, USA 8 Division of Molecular Histopathology, Department of Pathology, University of Cambridge, Cambridge, UK

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Anaplastic lymphoma kinase (ALK) inhibition has shown remarkable success in targeted therapy of non small cell lung cancer and in anaplastic large cell lymphoma (ALCL) where ALK kinase fusion proteins have initially been identified (ALK+ ) in 60 % of the patients. ALCL is a CD30 positive, aggressive Non-Hodgkin T-cell lymphoma mainly affecting children or young adults. Unfortunately treatment with ALK inhibitors inevitably leads to resistance development making the identification of new therapeutic targets an imperative. To achieve this we screened primary T-cell lymphoma samples and ALCL cell lines with a preselected library of 105 substances with known molecular targets and found that cells were highly susceptible to inhibitors of tyrosine kinase 2 (TYK2), a member of the Janus kinase family (JAK1, JAK2, JAK3, TYK2). In contrast, peripheral blood mononuclear cells (PBMC) and the JURKAT T-ALL cell line were not affected. In order to corroborate the importance of TYK2 for ALCL cell survival shRNA mediated gene knockdown was performed resulting in strong induction of apoptotic cell death. Analaysis of potential downstream targets (STAT1, STAT3, STAT5) revealed distinct p-STAT1 reduction after small molecule TYK2 inhibition. Accordingly, shRNA mediated gene knockdown of STAT1 revealed an essential function for ALCL cell survival. Taken together, our results indicate TYK2-pathway dependence in ALCL. Thus we identified a novel therapeutic intervention site that is druggable by recently developed small molecule inhibitors. Ongoing in vivo experiments using an ALCL transgenic mouse model in which TYK2 is knocked out in T-cells will systematically assess the therapeutic relevance of our findings.

 epartment of Internal Medicine III, Salzburg Cancer D Research Institute, Salzburg, Austria 2 Institute of Pathology, Paracelsus Private Medical University Salzburg, Salzburg, Austria 3 Institute of Radiology, Paracelsus Private Medical University Salzburg, Salzburg, Austria 4 Institute for Research and Development on Advanced RadiationTechnologies, Salzburg, Austria Introduction:  The management of metastatic cancer requires new specific, sensitive and ideally low invasive biomarkers to determine an early response to treatment. Circulating cell-free DNA (cfDNA) may represent such a non-invasive biomaker, as it can be found in higher concentrations in the plasma of cancer patients sharing some characteristics with DNA of tumor cells (cft-DNA) like specific oncogene or tumorsuppressor gene mutations. The aim of this work was to compare different established serum-biomarkers with our analysis of cfDNA and cft-DNA in correlation to disease monitoring by CT-imaging. Materials and Methods: We quantified the amount of cfDNA/cft-DNA in 15 patients with metastatic breast, colon or pancreatic cancer during their different treatments. We analyzed the ratio of level changes of cfDNA/cft-DNA and classical biomarkers, respectively, and compared these analysis to the disease phases as diagnosed by CT imaging according to RECIST criteria. Results:  We observed a relevant correlation of cfDNA (p < 0.0001) and cft-DNA (p = 0.0072) with RECIST defined response on longitudinal volumetric analysis of metastases in a subset of patients. In contrast to the excellent correlation at defined staging time-points, we observed a large signal of unexplained noise when analyzing samples obtained during treatment phases. In comparison to cfDNA/cft-DNA all other tumor marker did not show a consistent change in the quantitative values, corresponding the outcome of the CT-analysis. Conclusions:  Consecutive quantitative analyses of cfDNA and cft-DNA seem to be able to reflect the tumor burden during chemotherapy in patients with metastatic disease and pose an interesting field of current and future research.

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A06

Gene signatures of cancer stem cells and tumor bulk cells divide epithelial from neuroendocrine differentiation in a transgene mouse model of prostate cancer

ONCO-T-PROFIL: molecular characterization of potential predictive biomarkers for the treatment of patients with refractory metastatic solid tumours

Maximilian Marhold, Erwin Tomasich, Michael Schwarz, Simon Udovica, Gerwin Heller, Peter Horak, Michael Krainer

Andreas Seeber1, Guenther Gastl1, Christian Ensinger2, Verena Forcher2, Florian Rinderer1, Ella Willenbacher1, Wolfgang Willenbacher1, Wolfgang Eisterer1, Georg Pall1, Heinz Zwierzina1

 epartment of Medicine I, Comprehensive Cancer Center, D Medical Unversity of Vienna, Vienna, Austria Introduction:  Cancer Stem Cells (CSCs) have been described and characterized within prostate cancer (PCa) recently. During our previous work, we were able to identify and isolate basal CSC populations in a transgene mouse model of PCa. Materials and Methods:  Using fluorescence activated cell sorting (FACS), we sorted freshly isolated prostate tumor cells of Simian Virus-40 t-antigen driven TRAMP mice (Transgene Model of the Mouse Prostate) based on the expression of Sca-1 (Stem cell antigen 1) as well as the basal cell marker CD49f (Integrin α6) and used xenograft formation assays to confirm their stem and progenitor properties. Results:  RNA-Sequencing of the hence isolated cells identified distinct gene signatures for CSC and non-CSC subpopulations. Using this approach, we were able to show that non-CSCs include cells of neuroendocrine origin, whilst basal CSCs solely represent epithelial cells of origin for prostate cancer. Conclusions:  Our findings possibly explain, why neuroendocrine differentiation occurs during castration in one of the oldest and most frequently used animal models of PCa. Further, it might have translational value in explaining the development of castration resistance in the human disease.

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 epartment of Haematology and Oncology, Medical D University of Innsbruck, Innsbruck, Austria Department of Pathology, Medical University of Innsbruck, Innsbruck, Austria

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Introduction:  Patients with refractory metastatic cancer have been shown to benefit from molecular profiling of tumour tissue. The “ONCO-T-PROFIL” project was launched in March 2014 at the Department for Haematology and Oncology of Innsbruck Medical University. Within 2 years our project will recruit 100 patients with stage IV cancer. Our data presented here refer to one year of recruitment. Material and Methods:  Formalin-fixed tumour tissue specimens are submitted for molecular profiling to a certified laboratory in the USA (CARIS Life Intelligence©). Profiling methods used to identify and characterize potential predictive biomarkers include IHC, NGS and CISH. Druggable tumor targets and corresponding drug candidates and treatment regimes were selected based on an evidence-based information system that associates the biomarker status to agents with potential clinical benefit or potential lack of benefit. Clinical benefit was defined as a PFS ratio (= PFS upon the last therapy/PFS upon treatment according to the molecular profile) ≥ 1.3. Results:  Until January 2015, tumours from 31 patients have been molecularly profiled and in 30 (96.8 %) one or more druggable target structures were detectable. So far, 16 (51.6%) patients have been treated or are currently undergoing treatment according to their molecular tumor profile. To date, 5 of 16 patients have benefitted by reaching a PFS ratio of ≥ 1.3. Conclusions:  Up to now 16.1% of our patients showed a clinical benefit from a therapeutic regimen based on molecular typing. Our data demonstrate that a subgroup of patients can benefit from an individualized treatment approach based on molecular profiling.

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Klinische Studien

K01 AGMT_ERCC1 Pilotstudy: Biomarker directed treatment in metastatic colorectal cancer

Indication:  Inoperable locally advanced or metastatic gastric or gastro-esophageal junction (GEJ) cancer. Primary objective: Rate of HER2 positive locally advanced or metastatic gastric or GEJ cancer Secondary objectives: Comparison of HER2 results from local and central labs Safety of chemo-immunotherapy in HER2 positive patientsPFS and OS after chemo-immunotherapy Recruitment: 200—300 patients

Alois Lang1, Thomas Winder2  epartment of Internal Medicine E, LKH Feldkirch, Feldkirch, D Austria 2 Department of Oncology, University Hospital of Zurich, Zurich, Switzerland 1

EudraCT: 2011-003217-41 Start of study: Aug. 2012 End of recruitment: Q4 2015 Design:  This pilot study will investigate ERCC-1 (ExcisionRepair Cross-Complementing) as a predictive marker for treatment with platinum based regimens in mCRC patients. In a Pre-Screening phase the KRAS mutation status and ERCC-1 gene expression will be assessed at the CLIA approved laboratory Response genetics in Los Angeles, California, USA. KRAS wt patients will then be treated with 6 cycles of one of the following regimens chosen for optimization based on patient characteristics: ●● Patients with ERCC-1 < 1.7 relative gene expression of ERCC-1 over ß-actin (ERCC-1 low): mFOLFOX6 in combination with Cetuximab ●● Patients with ERCC-1 gene expression > 1,7 (ERCC-1 high): FOLFIRI in combination with Cetuximab The primary objective is to assess treatment response in patients with previous untreated wt KRAS advanced colorectal cancer using mFOLFOX6 or FOLFIRI and cetuximab with therapy chosen using ERCC-1 gene expression assessment. Recruitment (as of Jan. 2015): 26 of 50 patients

K02 AGMT_GASTRIC 5: PATIENT REGISTRY Screening for human epidermal growth factor receptor 2 (HER2) positivity in patients with inoperable locally advanced or metastatic gastric or gastro-esophageal junction (GEJ) cancer

K03 AGMT_HNO_Reg: PATIENTENREGISTER Prospektive Datenerhebung der Therapie bei Patienten mit lokal fortgeschrittenen und rezidivierenden/metastasierenden Kopf-Hals-Tumoren Richard Greil Universitätsklinik für Innere Medizin III, Paracelsus Medizinische Privatuniversität, Salzburg, Österreich Design:  Dieses Register ist als prospektiv beobachtende multizentrische Erhebung angelegt. Die Daten werden über einen Zeitraum von 10 Jahren in allen österreichischen Zentren erhoben, die ihre Zustimmung zur Teilnahme geben. Es wird mit einer Patientenzahl von etwa 500 gerechnet, das entspricht 50% der 1 Jahres-Österreich-Inzidenz. Für diese Dokumentation sind keine diagnostischen und therapeutischen Maßnahmen erforderlich, die über den ohnehin notwendigen und üblichen Rahmen hinausgehen. Durch die Dokumentation soll die Behandlungsroutine nicht verändert werden. In den letzten Jahren haben konsequent durchgeführte klinische Phase-IIIStudien eine Verbesserung der Heilungsraten bzw. Überlebenszeiten von Patienten in allen Stadien der Erkrankung gebracht. Dies ist auf adjuvante Chemoradiotherapieverfahren sowie insbesondere neo-adjuvante Triple Chemotherapien mit anschließender Chemoradiotherapie ± Operation zurückzuführen. Endpunkte:  Erhebung der Behandlungsrealität in Österreich, Sequenzen der Behandlung Stadien-spezifisch, Therapiewahl in den jeweiligen Behandlungslinien und Lokalisationen, Response-Evaluierung: Messung von RR, PFS und OS Einschluss:  Patienten mit eindeutig diagnostizierten Tumoren der Kopf-Hals-Region, Patienten, die mit dieser Datenerhebung ausdrücklich einverstanden sind Rekrutierung (Stand Jan. 2015): 421 von 500 Patienten

Ewald Wöll  epartment of Internal Medicine, St. Vinzenz Hospital, Zams, D Austria Design:  This registry is a prospective, observational, multicenter research initiative. In all eligible patients HER2 testing will be performed by means of IHC and in equivocal cases (Score 2+ ) in addition by ISH. HER2 positive and HER2 negative samples will be sent to a central pathology where a second HER2 testing will be performed. These test results will not influence the treatment of the individual patient, but will be analyzed retrospectively. All HER2 positive patients will receive further therapy at the discretion of the principal investigator. Kind of therapy and duration will be documented.

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Abstracts prior to the input of data. No informed consent is required from deceased patients.

K04 AGMT_HNO 2 Randomisierte Phase II Pilot Studie: Induktionstherapie mit Docetaxel, Cisplatin und Cetuximab versus Docetaxel, Cisplatin und 5 FU gefolgt von Radioimmuntherapie mit Cetuximab bei lokal fortgeschrittenen oder inoperablen Kopf-Hals Tumoren Felix Keil 3. Medizinische Abteilung, Hanusch Krankenhaus der Wiener Gebietskrankenkasse, Wien, Österreich EudraCT: 2011-005540-99 Start der Studie: Mai 2013 Rekrutierungsende: Q1 2015

Indication  ●● Histological evidence of breast cancer ●● Histological and/or radiological evidence of metastases ●● Metastasis within 10 years of registry initiation Primary objective: Epidemiological evaluations (general characteristics of metastatic stage patients in Austria, assessment of metastatic stage breast cancer subtypes in Austria, assessment of the specific characteristics and frequency of metastatic breast cancer, data on survival of female patients with metastatic breast cancer in Austria) and therapy-specific evaluations. Recruitment: 1000–3000 patients

K06

Design:  In dieser multizentrischen randomisierten PhaseII-Studie soll die Effizienz und Toxizität von TP plus Cetuximab mit TPF verglichen werden. Es soll geprüft werden, ob bei gleichbleibender Effizienz in einer taxanhältigen Induktionschemotherapie auf das 5FU verzichtet werden kann, da diese Substanz neben den Platinen ein erhebliches Potential für Schleimhauttoxizität hat. Speziell die Kombinationstherapie mit Docetaxel und Platinen ist im metastasierten Setting auch in Kombination mit Cetuximab hocheffektiv. Somit wäre eine Kombinationstherapie ohne 5FU für Patienten mit fortgeschrittenen HNO Tumoren eine sinnvolle Alternative. Alle PatientInnen erhalten 3 Zyklen (à 21 Tage) eine Induktions(immuno)chemotherapie. Im Anschluss startet vor Beginn der Radiotherapie (zumindest ein Tag) die Gabe von Cetuximab, die während der Gesamtdauer der Radiotherapie (konkomitante Boost-Radiotherapie über 6 Wochen) fortgeführt wird. Cetuximab wird während der Induktionschemotherapie im Arm B als Studienmedikation zur Verfügung gestellt. Bei den anderen in dieser Studie zum Einsatz kommenden Therapieformen und den dabei verwendeten Arzneimitteln handelt es sich um die in den jeweiligen Zentren etablierten Therapiemodalitäten bei gegebener Grunderkrankung. Primärer Studienendpunkt: Ansprechrate (CR, PR) 3 Monate nach Beendigung der Therapie Rekrutierung (Stand Jän. 2015): 73 von 100 Patienten

K05 AGMT_MBC_Reg: PATIENT REGISTRY Metastatic breast cancer in Austria Richard Greil Department of Internal Medicine III, Paracelsus Private Medical University, Salzburg, Austria

AGMT_MBC 6 Capecitabine in combination with Bendamustine in women with pretreated locally advanced or metastatic Her2-negative breast cancer, a Phase II Trial Richard Greil Department of Internal Medicine III, Paracelsus Private Medical University, Salzburg, Austria EudraCT: 2012-005593-64 Start of study: Aug. 2013 End of recruitment: Q1 2015 Design:  This is a non-randomized, multicenter, openlabel, single-arm Phase II study in pretreated patients with Her2-negative advanced breast cancer. Prior therapies must include anthracyclines and/or taxans in the adjuvant or metastatic setting. Following a two-stage design efficacy and safety of bendamustine and capacitabine will be evaluated following recruitment of the first 20 patients. Upon favorable results a further 20 patients will be recruited to reach the target population of 40 evaluable patients. Primary Endpoint of this study is to determine the efficacy of a capecitabine plus bendamustine combination regimen in the treatment of Her2-negative advanced metastatic breast cancer, in terms of overall response rates (complete or partial response, determined by radiologic evaluation according to Response Evaluation Criteria in Solid Tumors – RECIST Version 1.1 [23]. Patients:  40 eligible patients will be enrolled. A two-stage design efficacy and safety of bendamustine and capacitabine will be evaluated following recruitment of the first 20 patients. Upon favorable results a further 20 patients will be recruited to reach the target population of 40 evaluable patients. Recruitment (as of Jan. 2015): 32 of 40 patients

Design:  This registry is a prospective and retrospective, multi­center collection of data on patients with metastatic breast cancer in Austria. All tumor characteristics, medical histories and also treatment sequences are documented in anonymized form. For documentation in the registry, no further diagnostic or therapeutic measures are required than those already necessary in general. Participation in the registry must not interfere with treatment routines. A written consent must be obtained

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K07 ABCSG 36 (PENELOPEB) Phase-III-Studie zur Evaluierung von Palbociclib (PD-0332991) bei Patientinnen (HR+, HER2-normal) mit primärem Brustkrebs mit Rückfallrisiko nach neoadjuvanter Chemotherapie Michael Gnant, Gabriel Rinnerthaler Austrian Breast & Colorectal Cancer Study Group (ABCSG), Wien, Österreich Primäres Studienziel: Vergleich des invasiven krankheitsfreien Überlebens (iDFS) von Palbociclib mit Placebo bei Patientinnen mit invasivem Resttumor nach neoadjuvanter Chemotherapie und Residualtumor mit hohem CPS-EG Score, die eine adjuvante endokrine Standardtherapie bei HR-posi­ tivem/HER2-negativem primären Mammakarzinom erhalten. Sekundäre Studienziele:  Vergleich zwischen beiden Armen ●● iDFS (unter Nichtberücksichtigung der Zweitkarzinome) ●● Gesamtüberleben (OS) ●● Fernmetastasen-freies Überleben (DDFS) ●● Lokoregionäres rückfallfreies Überleben (LRRFS) ●● iDFS in Patienten mit reinen luminal-B Tumoren Studiendesign:  doppelblinde, randomisierte, prospektive, Placebo-kontrollierte Phase-III-Studie Therapie:  Adjuvante endokrine Standardtherapie kombiniert mit Palbociclib/Plazebo 13 Zyklen Wichtigste Einschlusskriterien: 1. Schriftliches Einverständnis vor Beginn jeglicher studienspezifischer Untersuchungen 2. Histologisch bestätigtes unilaterales/bilaterales primäres invasives Karzinom der Brust 3. Einverständnis und Möglichkeit der Bereitstellung eines gelagerten Formalin-fixierten Paraffin-eingebetteten Gewebeblocks (FFPE) oder eines Teilblocks von der Oper­ation nach neoadjuvanter Chemotherapie und einer Stanzbiopsie vor Start der neoadjuvanten Chemotherapie 4. Invasiver Resttumor nach der Chemotherapie, entweder in der Brust und/oder den Lymphknoten 5. Zentral bestätigter HR+  und HER2-normal Status 6. Adäquate Operation mit Resektion des gesamten Rest­ tumors und ipsilateraler axillarer Lymphknotendissektion. Histologische Komplettresektion (R0) des invasiven und nicht-invasiven (DCIS) Tumors notwendig (falls brusterhaltende Operation als finale Behandlung). Axilläre Dissektion bei Patientinnen mit negativer Sentinellymph­knotenbiopsie vor (pN0,pN+ (mic)) oder nach (ypN0, ypN+ (mic)) neoadjuvanter Chemotherapie nicht notwendig. Wichtigste Ausschlusskriterien: 1. Bekannte schwere Hypersensitivitätsreaktionen gegen Präparate ähnlich Palbociclib oder Palbociclib/PlaceboHilfsstoffen oder gegen die endokrine Behandlung 2. Inadäquate Organfunktionen unmittelbar vor der Randomisierung definiert durch bestimmte Blutparameter 3. Akute schwere oder unkontrollierte, systemische Erkrankungen.

K08 ABCSG 38 (LORELEI) Eine randomisierte, doppelblinde Phase-II-Studie zum Vergleich von neoadjuvantem Letrozol plus GDC-0032 versus Letrozol plus Placebo bei postmenopausalen Frauen mit ER-positivem/HER2-negativem Brustkrebs im Frühstadium Michael Gnant, Peter Dubsky, Gabriel Rinnerthaler Austrian Breast & Colorectal Cancer Study Group (ABCSG), Wien, Österreich Primäre Studienziele: Evaluierung der Wirksamkeit von Letrozol plus GDC-0032 im Vergleich zu Letrozol plus Placebo bei Frauen mit ER-positivem/ HER2-negativem, primärem Brustkrebs, mittels: ●● Gesamter Tumor ORR, zentral erhoben mittels Kernspinresonanz Bildgebung und ausgewertet mittels modifizierter „Response Evaluation Criteria in Solid Tumors“ (RECIST) bei allen eingebrachten Patientinnen und bei PIK3CA mutierten (MT) Patientinnen. ●● pCR Rate in Brust und Axilla bei allen eingebrachten Patientinnen und bei PIK3CA mutierten (MT) Patientinnen, basierend auf lokalen Auswertungen. Sekundäre Studienziele: Erhebung der ORR bei PIK3CA Wild Typ (WT) Patientinnen zentral erhoben mittels Kernspinresonanz Bildgebung und ausgewertet mittels modifizierter „Response Evaluation Criteria in Solid Tumors“ (RECIST), sowie die lokale Erhebung der pCR Rate, bei diesem Patientenkollektiv. Studiendesign:  doppelblinde, randomisierte Phase II Studie Therapie:  Arm A: 16 Wochen Letrozol + GDC-0032Arm B: 16 Wochen Letrozol + GDC-0032 Placebo Wichtigste Einschlusskriterien: 1. Unterschriebene Einwilligungserklärung vor Durchführung von studienrelevanten Handlungen 2. Weibliches Geschlecht 3. Postmenopausal und ≥ 18 Jahre 4. Histologisch bestätigtes, invasives Mammakarzinom mit allen der folgenden Charakteristika: ER-positiv und HER2-negativer Brustkrebs gemäß lokaler Laborauswertung oder lokalen Bestimmunge Wichtigste Ausschlusskriterien: 1. Vorangegangene Behandlung eines primär invasiven Mammakarzinoms 2. Patientinnen mit Brusttumoren im Stadium cT4 oder cN3 3. Metastasierter Brustkrebs (Stadium IV) 4. Bilateral invasives/ multizentrisches Mammakarzinom Rekrutierungszeitraum:  bis Q4 2016

Rekrutierungszeitraum: bis Q4 2016

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K10

ABCSG 41 (OLYMPIA) Doppelblinde, Placebo-kontrollierte Studie zur Beurteilung der Wirksamkeit sowie der Unbedenklichkeit des PARP-Inhibitors Olaparib als adjuvante Behandlung von triple negativen BrustkrebsPatientinnen mit BRCA1/2-Keimbahnmutation

Cardiovascular Complications of Cancer Treatment: Update CACOCA Trial

Christian Singer, Gabriel Rinnerthaler

 epartment of Internal Medicine V, Medical University of D Innsbruck, Innsbruck, Austria 2 Tyrolean Cancer Research Institute, Innsbruck, Austria 3 Department of Internal Medicine III, Medical University of Innsbruck, Innsbruck, Austria 4 Department of Internal Medicine, Hospital Bregenz, Bregenz, Austria 5 Department of Internal Medicine III, General Hospital Linz, Linz, Austria 6 Department of Obstetrics and Gynecology, Medical University of Innsbruck, Innsbruck, Austria 7 Department of Internal Medicine, Hospital Feldkirch, Feldkirch, Austria 8 Deprtment of Internal Medicine I, Hospital BHS Linz, Linz, Austria 9 Department of Internal Medicine, Hospital Zams, Zams, Austria 10 Department of Internal Medicine I, Wilhelminenspital Wien, Vienna, Austria

Austrian Breast & Colorectal Cancer Study Group (ABCSG), Wien, Österreich Primäre Studienziele: Beurteilung der Wirksamkeit der adjuvanten Behandlung mit Olaparib für das invasive krankheitsfreie Überleben („Invasive Disease Free Survival“, IDFS). Bei primärem HER2-negativem Hochrisiko-Brustkrebs Patienten, welche eine BRCA1/2-Keimbahnmutation aufweisen und eine definitive lokale Behandlung und neoadjuvante bzw. adjuvante Chemotherapie abgeschlossen haben. Sekundäre Studienziele:  Beurteilung der Wirksamkeit der adjuvanten Behandlung mit Olaparib, für das Gesamtüberleben („Overall Survival“, OS), das fernmetastasenfreie Überleben („Distant Disease Free Survival“, DDFS). Beurteilung der Wirksamkeit des Prüfpräparats im Falle des Auftretens von neuem invasiven primären Brustkrebs und/oder einem neuen Ovarialkarzinom im Epithelgewebe. Studiendesign:  doppelblinde, randomisierte, Placebokontrollierte Phase III Studie Therapie:  Arm A: 12 Monate Olaparib Arm B: 12 Monate Placebo Wichtigste Einschlusskriterien:  1. Schriftliches Einverständnis vor Beginn jeglicher studienspezifischer Untersuchungen 2. Weibliche und männliche Patienten: ≥ 18 Jahre 3. Histologisch bestätigtes, nicht-metastasiertes, primäres, triple-negatives (*Protokollamendment in Planung: Einschluss von ER- und PR-positiven PatientInnen), invasives Adenokarzinom der Brust, das bei der Operation: • entweder positive Lymphknoten (jede Tumorgröße) zeigt oder negative Lymphknoten mit einem Primärtumor > 2 cm bei PatientInnen, die adjuvante Chemotherapie erhalten haben oder • nachgewiesene non-pCR bei PatientInnen, die neoadjuvante Chemotherapie erhalten haben 4. Bestätigte BRCA-Mutation 5. Abgeschlossene neoadjuvante oder adjuvante Chemotherapie (min. 6 Zyklen) mit Anthrazyklinen, Taxanen oder einer Kombination von beiden. Wichtigste Ausschlusskriterien: 1. Metastasierter Brustkrebs 2. Vorhergehende Therapie mit einem PARP-Inhibitor 3. Andere Krebserkrankungen (Ausnahme: ausreichend behandelte Basalzellkarzinome der Haut und/oder in-situ-Karzinome der Cervix)

Florian Kocher1,2, Stephan Dobner3, Bernhard Föger4, Michael Fridrik5, Georg Pall1, Michael Hubalek6, Alois Lang7, Andreas Petzer8, Ewald Wöll9, Wolfgang Hilbe1,10 1

Introduction:  Oncological treatment, applying chemotherapy or new targeted drugs has already shown to induce cardiotoxicity. Present knowledge is primarily based on patients, who were treated in prospective clinical trials and is focused on acute toxicities. In contrast, there is a lack of data concerning the long-term cardiotoxic effects of systemic cancer therapies. The present trial aims to prove the risk and the consequences of long-term cardiotoxicity experienced after different cancer treatments in an unselected cohort of patients. Methods:  In this prospective, non-interventional, multicenter trial 1000 patients suffering from breast cancer, color­ ectal cancer, non-small cell lung cancer or lymphoma will be included. Curative intervention (operation and/or radiotherapy) has to be completed and patients must qualify for adjuvant systemic treatment. In case of lymphoma, stage and histology must offer the option of a long term remission after systemic therapy. Patients will be evaluated in regard to their cardio­ vascular status, risk factors and quality of life over a time period of 60 months. Objectives:  Primary objective is to evaluate the incidence of major cardiac events defined as cardiac related death, severe cardiac symptoms or cardiac disorders which require invasive interventions. Secondary objectives are the incidence of patients with minor cardiac events, PFS, OS, QOL, physical performance status and biomarker assessment. Current Status:  The first patient was enrolled in October 2013. In January 2015, 10 centers have been initiated and 118 patients were included in the study. Further centers will be invited to participate.

Rekrutierungszeitraum:  bis Q1 2018

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K11 AGMT_ALCL1 A “window of opportunity” trial with Brentuximab Vedotin and Imatinib in patients with relapsed or refractory ALK+ anaplastic large cell lymphoma or patients ineligible for chemotherapy Ulrich Jäger Division of Hematology and Hemostaseology, Department of Medicine I, Medical University of Vienna, Vienna, Austria EudraCT: 2013-003505-26 Start of study: Q2 2015 End of recruitment: Q4 2016 Design:  This is an open label pilot study of combining BV in a licensed indication with imatinib in patients with ALCL. It is intended as a “window of opportunity” trial in which the study drugs will be given as an initial substitute for conventional chemotherapy with the intention to achieve a remission enabling the patients to proceed to autologous or allogeneic stem cell transplantation, if eligible. Imatinib will be given continuously starting from day 1 of the first cycle at an oral dose of 100mg daily. The dose will be increased to 200mg daily starting from day 1 of the second cycle if no DLT occurs during the first cycle. BV will be given 3 weekly starting on day 1 at a dose of 1.8 mg/kg body weight. In the absence of a dose limiting toyicity (DLT) after 3 weeks of therapy, and in the presence of a clinical response (CR or PR) after cycle 1, the BV dose will continue every 3 weeks for 48 weeks. Primary Endpoint of this study is to determine the safety and tolerability of simultaneous administration of brentuximab vedotin and imatinib mesylate in substitution of conventional chemotherapeutic treatment. Patients:  A total of 10 patients with relapsed or refractory ALK-positive ALCL will be included. Patients must have received at least one prior line of conventional chemotherapy for ALCL. Patients who are not eligible for conventional chemotherapy can also be included.

K12 AGMT-ALL Reg: Patient registry – Registry and biobank for the collection of clinical data and biomaterial from adult ALL patients Ulrich Jäger, Alexander Hauswirth Division of Hematology and Hemostaseology, Department of Medicine I, Medical University of Vienna, Vienna, Austria Design:  In Austria approximately 70 patients are diagnosed with adult ALL per year and are treated in up to 17 institutes. Obviously there is a need to collect all data possible in order to harmonize diagnosis and treatment and to make optimal therapy available for every Austrian patient. Therefore information on these patients should be prospectively collected, analysed and used for the generation of treatment protocols by a specialized study group. The Austrian ALL study group has been founded to take over this task. As a first step a registry with a standardized data set including diagnosis, therapy and outcome should

be implemented. This study is a registry and not an interventional clinical study. However, in order to achieve a maximum of data harmonisation it is recommended that patients are treated according to a standardized international protocol endorsed by the EWALL (European Working group for Adult Lymphoblasitc Leukemia). Associated with the registry a central biobank should be established for the collection of peripheral blood and bone marrow. During the last years the utility of molecular cytogenetics in adult ALL is an emerging topic. Several prognostic markers have been discovered and new targeted drugs are available. The use of molecular diagnosis in disease monitoring, risk stratification and the use of target orientated therapies are increasingly important in patient management of ALL. These diagnosis tools are currently not implemented in Austria. For that reason there are many open questions in adult ALL and the new prospects leave clinicians with uncertainty about how to optimally manage adult patients with ALL. A centralized and standardized diagnosis program is needed to assure quality. Objectives:  Data collection regarding diagnosis, therapy and progression of disease for Austian ALL patients older than 18 years. Biobank:  Within the ALL registry biomaterial should be collected at diagnosis and once a year for 5 years and at relapse.

K13 HD16 für frühe Stadien: Therapieoptimierungsstudie in der Primärtherapie des frühen Hodgkin Lymphoms: Therapiestratifizierung mittels FDG-PET Richard Greil Universitätsklinik für Innere Medizin III, Paracelsus Medizinische Privatuniversität, Salzburg, Österreich EudraCT: 2007-004474-24 Phase III Studie in Kooperation mit der GHSG Köln Studienstart in Österreich: März 2012 Rekrutierungsende: Q2 2015 Design:  ABVD ist aufgrund der hervorragenden Tumorkontrolle bei gleichzeitig niedriger Toxizität das in der GHSG etablierte Schema für HL-Patienten in frühen Stadien. Die Frage der Äquivalenz von 2 x ABVD und 2 x AVD konnte mit der Endauswertung der HD13 Studie nicht bestätigt werden. Aufgrund der Ergebnisse der HD10-Studie stellen 20 Gy IF-RT im Rahmen von Studien den neuen Standard dar. In der beschriebenen HD16 Studie wird mittels der FDG-PET nach 2 Zyklen Chemotherapie das frühe Ansprechen auf die Chemotherapie untersucht. Patienten, die in den Standardarm randomisiert werden, erhalten unabhängig vom Ergebnis der PET-2 eine IF-RT (20 Gy). Patienten, die in den experimentellen Arm randomisiert werden, erhalten bei einem positiven PET (PET-2 positiv) eine IF-RT (20 Gy). Patienten mit einer negativen PET (PET-2 negativ) erhalten keine weitere Therapie. Der primäre Endpunkt ist die Dauer des „progressionsfreien Überlebens“ (Progression Free Survival, PFS). PFS als Hauptendpunkt reflektiert das Tumorwachstum und eignet sich daher besonders gut für einen Armvergleich, bei dem der Armunterschied nach anfänglich gutem Ansprechen auf die Chemotherapie wirksam wird. Rekrutierung (Stand Jän. 2015): 34 Patienten in Ö (975 von 1100 internat.)

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K14 HD17 für intermediäre Stadien: Therapieoptimierungsstudie in der Primärtherapie des intermediären Hodgkin Lymphoms: Therapiestratifizierung mittels FDG-PET Richard Greil Universitätsklinik für Innere Medizin III, Paracelsus Medizinische Privatuniversität, Salzburg, Österreich EudraCT: 2007-005920-34 Phase III Studie in Kooperation mit der GHSG Köln Studienstart in Österreich: Jänner 2013 Rekrutierungsende: Dezember 2016 Design:  Der Therapiestandard für Patienten mit Erstdiagnose eines Hodgkin Lymphoms in intermediärem Stadium ist nach den aktuellen Ergebnissen der HD 14 Studie unserer Studiengruppe eine kombinierte Chemo- und Radiotherapie bestehend aus 2 Zyklen BEACOPP eskaliert und 2 Zyklen ABVD gefolgt von einer 30 Gy IF-Strahlentherapie. Die Behandlungsergebnisse von Patienten in intermediären Stadien sind hiermit bezüglich des progressionsfreien Überlebens und des Gesamtüberlebens bereits sehr gut. In der HD17 Studie wird mittels der FDG-PET nach 4 Zyklen Chemotherapie (PET-4) das Ansprechen auf die Chemotherapie untersucht. Patienten, die in den Standardarm randomisiert werden, erhalten unabhängig vom Ergebnis der PET-4 eine IF-RT (30 Gy „involved field“). Patienten, die in den experimentellen Arm randomisiert werden, erhalten bei einem positiven PET (PET-4 positiv) eine IN-RT (30 Gy „involved node“). Patienten mit einer negativen PET (PET-4 negativ) erhalten im experimentellen Arm keine weitere Therapie. Die bewährte IF-Radiotherapie mit 30 Gy wird in der vorliegenden Studie mit einer interessanten Neuentwicklung im Bereich der Strahlentherapie hinsichtlich der Zielvolumen­ definition des „involved node“ verglichen. Eine weitere Minimierung von Akut- und Spättoxizität der Radiotherapie bei gleicher Effektivität wird erwartet. Darüber hinaus wird die Studie zeigen, ob man bei PET negativen Patienten nach der Chemotherapie möglicherweise auf eine Bestrahlung verzichten kann. Primärer Endpunkt: PFS Rekrutierung (Stand Jän. 2015): 25 Patienten in Ö. (644 von 1100 internat.)

K15 AGMT_ HMA in Myeloid Neoplasms: Registry on hypomethylating agents in myeloid neoplasms, including myelodysplastic syndrome (MDS), CMML and AML Richard Greil, Lisa Pleyer Department of Internal Medicine III, Paracelsus Private Medical University, Salzburg, Austria Design:  This registry is a retrospective and prospective, observational, multi-center research initiative. Data will be col-

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lected from all sites willing to participate. It is expected, that the main data-bulk will be obtained from approximately 20 sites. An estimated 800–1000 patients are expected to be included; these numbers may be revised over time as interest and demand dictates. No pre-defined visits, medical tests, laboratory tests, procedures, or interventions are required. Physicians who have already treated patients with hypomethylating agents or are planning to initiate treatment with a hypomethylating agent can include patient data in this registry. To help maintain patient confidentiality, each patient will be assigned a unique patient identifying number upon enrollment; this number will accompany the patient’s medical and other registry information throughout the lifetime of the registry. Additionally from some patients with myeloid neoplasms, blood or tissue samples will be stored for further analyses. These samples will also be obtained from patients, who are not or not yet treated with hypomethylating agents to comprise a control. Inclusion:  Begin with or already have received treatment with a hypomethylating agent. Be willing to provide informed consent. The objectives of the study are number of cycles and dosage of VIDAZA® therapy, response evaluation, toxicities, severe adverse reactions, overall survival. Recruitment (as of Jan. 2015): 661 patients in Austria

K16 NHL7-2008/A: Prospektiv randomisierte Studie zur Therapieoptimierung (Primärtherapie) fortgeschrittener progredienter follikulärer und anderer niedrig maligner sowie Mantelzell Lymphome Michael Fridrik JKU Forschungszentrum, Allgemeines Krankenhaus Linz, Linz, Österreich EudraCT: 2008-005859-16  ekrutierung nur mehr für Morbus Waldenström Patienten R offen! Phase-III-Studie in Kooperation mit der StiL Forsc­hungsgruppe Studienstart in Österreich: Okt. 2009 Rekrutierungsende: 2017 Design:  Im Rahmen dieser Studie soll die Wertigkeit der Dauer der Erhaltungstherapie mit Rituximab in der Erstlinientherapie der follikulären Lymphome untersucht und folgende Frage beantwortet werden: Kann mit einer längeren zielgerichteten antilymphoproliferativen Erhaltungstherapie mit Rituximab (4 Jahre im Vergleich zu 2 Jahre) eine Eradikation der malignen Lymphomzellen, also eine Heilung der Erkrankung bzw. eine längere progressionsfreie Zeit erreicht werden. Bei den anderen Lymphomentitäten Immunozytom, Marginalzonen und Mantelzell Lymphom soll die Frage beantwortet werden, ob die Prognose dieser Erkrankungen durch eine verlängerte Rituximab Therapie im Vergleich zu der sonst üblichen therapiefreien Nachbeobachtung verbessert werden kann (2  Jahre Erhaltungstherapie im Vergleich zu Nachbeobachtung). Primäres Untersuchungsziel beim Immunozytom:  Ermittlung und Vergleich des progressionsfreien Überlebens beider Therapien Bendamustin plus Rituximab ohne Erhaltungsthera-

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Abstracts pie versus Bendamustin plus Rituximab und 2 Jahre Rituximab Erhaltungstherapie. Rekrutierung (Stand Jän. 2015 in Ö.): 68 Patienten (19.  M Waldenström)

K17 AGMT_PTCL-Reg: Patientenregister – Erhebung epidemiologischer Daten von T-Zell Lymphomen in Österreich Richard Greil Universitätsklinik für Innere Medizin III, Paracelsus Medizinische Privatuniversität, Salzburg, Österreich Design:  Dieses Register ist eine prospektiv und retrospektiv beobachtende, multizentrische Erhebung. Die Daten werden in allen österreichischen Zentren erhoben, die ihre Zustimmung zur Teilnahme geben. Für diese Dokumentation sind keine diagnostischen und therapeutischen Maßnahmen erforderlich, die über den ohnehin notwendigen und üblichen Rahmen hinausgehen. Durch die Dokumentation soll die Behandlungsroutine nicht verändert werden. Um die Vertraulichkeit der Daten aller Patienten zu gewährleisten, wird jedem Patienten zu Beginn der Datenerhebung eine Nummer zugeordnet, welche für die gesamte Laufzeit des Registers seine Gültigkeit behält. Die Sammlung der Daten erfolgt über ein elektronisches Datenerfassungssystem. Zusätzlich soll im Rahmen des Registers von allen Patienten, die dazu Ihr Einverständnis geben, Biomaterial gesammelt und zentral gelagert werden, um eine Basis für zukünftige wissenschaftliche Untersuchungen von krankheitsspezifischen Parametern zu schaffen. Endpunkte:  Verteilungsmuster (2008-WHO-Klassifikation), Therapien, Ansprechen. Einschluss:  Patienten mit einem peripheren T-Zell-Lymphome (PTCL) laut 2008-WHO-Klassifikation. Patienten, die mit dieser Datenerhebung ausdrücklich einverstanden sind. Rekrutierung (Stand Jän. 2015): 31 Patienten

K18 Phase 1 study to evaluate the feasibility and efficacy of the addition of P1101 to imatinib treatment in patients with CP-CML not achieving a CMR Sonja Burgstaller1, Michael Steurer2, Richard Greil3, Josef Thaler1  epartment of Internal Medicine IV, Klinikum D Wels-Grieskirchen, Wels, Austria 2 Department of Internal Medicine V, Medical University of Innsbruck, Innsbruck, Austria 3 Department of Internal Medicine III, Paracelsus Private Medical University, Salzburg, Austria 1

Introduction:  Mainly based on in vivo evidence imatinib alone is not able to eradicate the whole BCR-ABL clone. In the majority of CML patients BCR-ABL transcripts are still detectable even after years of imatinib treatment. The addition of P1101, a new PEG-Proline-Interferon alpha-2b, to current ima-

tinib treatment is expected to deepen molecular remission and increase the rate of complete molecular remission. Treatment:  Patients with chronic phase chronic myeloid leukaemia treated with imatinib for at least 18 months are eligible for study participation. Achievement of CHR and CCyR are mandatory before entering the study. Imatinib will be continued at the same dose level as before study entry. Dose modifications are limited to interferon therapy. P1101 will be added in a dose of 50 µg subcutaneously every 14 days. In the absence of a dose limiting toxicity (DLT) at 3 months the dose of P1101 will be increased to 100 µg subcutaneously every 14 days. DLT is defined as haematological toxicities grade ≥ 2 or non haematological toxicities grade ≥ 3. 100 µg P1101 is considered as the maximum dose, further increase of P1101 is not planned. Primary endpoint:  To determine the satety and tolerability of the additon of P1101 to the currently established dose of imatinib Secondary endpoint: To determine the rate of achievement of ≥ 1 log reduction from the initial BCR-ABL transcript level at study entry and the achievement of molecular remission 4.5 or undetecable BCR-ABL transcripts. First patient in Q2 2013, last patient out Q4 2015.

K19 AGMT_MM1: Ixazomib in Kombination mit Thalidomid – Dexamethason bei Patienten mit reapsiertem und/oder refraktärem multiplen Myelom Heinz Ludwig Wilhelminen Krebsforschungsinstitut c/o 1. Medizinische Abteilung - Zentrum für Onkologie, Hämatologie und Palliativmedizin, Wilhelminenspital, Wien, Österreich 

EudraCT: 2014-002749-23 Start of study: Q2 2015 End of recruitment: Q2 2017 Design:  This is an open phase II, single-arm, multi-center study to evaluate progression free survival in patients receiving ixazomib in combination with thalidomide and dexamethasone (ITD) followed by an ixazomib maintenance phase of a maximum period of 12 months. The patient population will consist of adult male and female patients with multiple myeloma (MM) with relapsed and/or refractory disease after at least one prior treatment line. Therapy regime: Ixazomib: 4.0 mg on days 1, 8, 15 Thalidomide: 100 mg on days 1–28 Dexamethasone: 20/40 mg on days 1, 8, 15 The primary endpoint is progression free survival (PFS), defined as the time from day 1 of cycle 1 to the date of first documentation of disease progression based on IMWG criteria or death due to any cause, whichever occurs first. Patients: At least 77 patients with at least 2 study cycles co­mpleted will be evaluated for statistical analyses in the study. 14 study sites in Austria, Germany and Czech Republic are planned to be recruited for the study.

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P02

Poster Hämatologie

P01 The Ki-1 antigen (CD30), a novel target in neoplastic canine mast cells, is downregulated by interleukin-4 Karin Bauer1, Emir Hadzijusufovic1,2,3, Sabine Cerny-Reiterer1,2, Armin Pirker4, Martin Reifinger5, Peter Valent1,2, Michael Willmann3  epartment of Internal Medicine I, Medical University of D Vienna, Vienna, Austria 2 Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria 3 Clinical Unit of Internal Medicine, University of Veterinary Medicine Vienna, Vienna, Austria 4 Clinical Unit of Surgery, University of Veterinary Medicine Vienna, Vienna, Austria 5 Department of Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria 1

The Ki-1 antigen (CD30) is a novel target in human mast cell (MC) neoplasms. However, so far, CD30 has not been investigated in the context of canine MC tumors. Here, we examined the expression, regulation and function of CD30 in neoplastic canine MCs. The canine mastocytoma cell lines C2 and NI-1 expressed CD30 as determined by immunocytochemistry. Flow cytometry confirmed cell surface expression of CD30 on canine MCs with higher levels found in NI-1 cells compared to C2 cells. To study the regulation of CD30 expression, we applied a panel of immunomodulatory cytokines. Interestingly, of all cytokines tested, interleukin-4 was found to downregulate expression of CD30 on canine MCs. Next, we asked whether CD30 can serve as a therapeutic target in mastocytomas. We found that the CD30-targeting antibody brentuximab vedotin induces growth inhibition in canine MCs, with higher IC50 values obtained in NI-1 cells compared to C2 cells. These growth inhibitory effects of brentuximab vedotin were associated with induction of apoptosis. Additionally, incubation of canine mastocytoma cell lines with brentuximab vedotin as well as with the tyrosine kinase inhibitors masitinib and PKC412 resulted in a significant and dose-dependent decrease in CD30 surface expression on NI-1 cells and C2 cells. Furthermore, brentuximab vedotin was found to synergize with masitinib and PKC412 in inhibiting the proliferation of NI-1 cells and C2 cells. Together, we have identified CD30 as a target of neoplastic canine MCs. Whether CD30-targeting drugs are effective in canine mastocytoma patients, remains to be determined.

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CAR, a novel marker of erythroid differentiation and migration, is downregulated in erythropoietic progenitor cells in MDS Karin Bauer1, Sigrid Machherndl-Spandl2, Susanne Suessner3, Martin Danzer3, Johannes Proell3, Jeroen Lauf4, Gregor Eisenwort1,5, Wolfgang R. Sperr1,5, Hans-Ulrich Klein6, Gregor Hoermann7, Norman Häfner8, Marie C. Béne9, Uwe Platzbecker10, Ansgar Weltermann2, Christian Gabriel3, Thomas Lion11, Ulrich Germing12, Peter Bettelheim2,4, Peter Valent1,5  epartment of Internal Medicine I, Medical University of D Vienna, Vienna, Austria 2 Division of Hematology and Oncology, Elisabethinen Hospital Linz, Linz, Austria 3 Department of Molecular Biology, Transfusion Service of Upper Austria, Linz, Austria 4 Labor Europaplatz, Linz, Austria 5 Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria 6 Institute of Medical Informatics, University of Muenster, Muenster, Germany 7 Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria 8 Department of Gynaecology and Obstetrics, Jena University Hospital, Jena, Germany 9 Hematology Laboratory, CHU De Nantes, Nantes, France 10 Division of Hematology, University of Dresden, Dresden, Germany 11 Children´s Cancer Research Institute (CCRI) Vienna and Department of Pediatrics, Medical University Vienna, Vienna, Austria 12 Department of Hematology, Oncology and Clinical Immunology, Heinrich-Heine-University, Düsseldorf, Germany 1

Myelodysplastic syndromes (MDS) are myeloid neoplasms characterized by peripheral cytopenia and by an accumulation of dysplastic erythroid progenitor cells (EryPC) in the bone marrow (BM). To examine abnormally regulated genes expressed in EryPC, we have established a screening program involving mRNA expression profiling of EryPC in patients with low-risk MDS and control BM samples. EryPC were defined as CD45low/ CD105+  cells and purified from BM cells by cell sorting. We found that the major Coxsackie-Adenovirus-Receptor (CAR) is specifically downregulated in EryPC in MDS patients when compared to EryPC in normal BM or other control cohorts. In line with this observation, the immature erythroblastic cell lines HEL, K562 and KU812 stained negative for CAR. Lentiviral transduction of the full-length CAR gene into these cells resulted in a significantly increased expression of various erythroid differentiation antigens, including CD36, CD71 and Glycophorin-A. Furthermore, CAR transduction resulted in an increased migration of HEL cells against a serum protein-gradient in a transwell-assay. Transfection with truncated variants of CAR did not result in an increased expression of erythroid antigens or increased migration. In conclusion, our data show that CAR is a functionally relevant molecule that promotes the expression of early erythroid differentiation antigens on myeloid progenitors and their migration against blood serum proteins. In patients with MDS, CAR is downregulated on EryPC, which may have implications in clinical diagnostics and may explain the maturation-defect of EryPC in MDS and the related accumulation of erythroid cells in the BM that is accompanying the peripheral anemia in these patients.

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P03 The pan-BCL-2 blocker obatoclax (GX15-070) and the PI3K/mTOR-inhibitor BEZ235 exert growth-inhibitory effects on neoplastic cells in Ph+ and Ph- ALL Daniela Berger1, Gabriele Stefanzl1, Katharina Blatt1, Gregor Eisenwort1,2, Wolfgang Sperr1, Alexander Hauswirth1, Ulrich Jäger1, Sabine Cerny-Reiterer1,2, Peter Valent1,2  epartment of Internal Medicine I, Medical University of D Vienna, Vienna, Austria Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria

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2

Acute lymphoblastic leukemia (ALL) is characterized by abnormal expansion of lymphoblasts in hematopoietic tissues. Despite improved therapy only a subset of patients can be cured. Therefore, current research is focusing on new drug targets and novel targeted drugs to improve therapy. Members of the BCL-2 family and components of the PI3K/mTOR pathway are critically involved in the regulation of growth and survival of ALL cells. We therefore examined the effects of the panBCL-2 blocker obatoclax (GX15-070) and the dual PI3K/mTOR blocker BEZ235 on growth and survival of ALL cells. As assessed by 3H-thymidine uptake, both drugs were found to suppress in vitro proliferation of leukemic cells in patients with Ph+ ALL (n = 4) and Ph- ALL (n =9) in a dose-dependent manner (obatoclax IC50: 0.05-0.1 µM; BEZ235 IC50: 0.01-0.5 µM). Both drugs also suppressed growth in Ph+ cell lines (BV-173, NALM-1, TOM-1, Z-119) and Ph- cell lines (RAJI, RAMOS, REH, BL-41). Moreover, obatoclax and BEZ235 induced apoptosis in all cell lines tested as evidenced by Western blotting using an antibody against cleaved caspase 3. In drug-combination experiments, obatoclax was found to cooperate with BEZ235 in inhibiting growth and survival of ALL cells. Finally, we were able to show that primary ALL cells, including CD34+ /CD38- ALL stem cells, and all cell lines tested, express transcripts for PI3K, mTOR, BCL-2, MCL-1 and BCL-xL by qPCR. Together, our data show that obatoclax and BEZ235 are novel promising drugs that block the growth and survival of Ph+ and Ph- ALL cells.

P04 Tryptase screening in 1330 unselected cases evaluated in a hematology-center reveals a cohort with hypertryptasemia of uncertain significance Katharina Blatt1, Susanne Herndlhofer1, Gregor Hoermann2, Wolfgang R Sperr1,3, Peter Valent1,3  epartment of Internal Medicine I, Medical University of D Vienna, Vienna, Austria 2 Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria 3 Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria 1

In systemic mastocytosis (SM) and other myeloid neoplasms, patients often have a slightly elevated serum tryptase

(15-30 ng/ml). However, an increased tryptase may also be found in allergic patients and sometimes even in healthy individuals. We determined tryptase-levels in 1330 individuals, including 914 patients with hematological neoplasms (myeloproliferative neoplasms, n =156; myelodysplastic syndromes, MDS, n =241; acute myeloid leukaemia, AML, n =317; SM, n =81; lymphomas, n =59; acute lymphoblastic leukaemia, n =26), 136 with non-neoplastic hematological disorders, 102 with nonhematological conditions, and 178 healthy subjects. In healthy individuals, the median serum tryptase-level amounted to 5.2 ng/ml. Elevated tryptase-levels were primarily found in myeloid neoplasm (SM: 90% of patients; AML: 38%; MDS: 25%). We also identified 15 cases (1.2%) in whom tryptase-levels were slightly elevated, but neither an allergic disease nor a hematologic disease could be identified. The median age of these 15 individuals was 44 years (range: 24-72 years) and the femaleto-male ratio was 2:1. The median tryptase-level at first presentation was 20.6 ng/ml (range: 14.5-35.5 ng/ml). In all cases, tryptase-levels remained stable in the follow up. Bone marrow examinations did not reveal any signs of a myeloid neoplasm, and in none of the cases, an abnormal karyotype or a KIT mutation at codon 816 were detected. Blood counts and differential counts were normal in all cases, and no molecular aberrations were detected by multiplex PCR. Together, we have identified a cohort of apparently healthy individuals with a persistently elevated tryptase-level. We propose to call this condition hypertryptasemia of unknown (uncertain) significance.

P05 Hairy cell leukemia – a retrospective analysis with regard to epidemiology and clinical outcome Jan-Paul Bohn, Gabriele Gamerith, Eberhard Gunsilius, Günther Gastl, Michael Steurer Department of Internal Medicine V (Hematology and Oncology), Medical University of Innsbruck, Innsbruck, Austria Introduction:  Hairy cell leukemia (HCL) is an uncommon, but distinct chronic B-cell-malignancy. Here, we report on the epidemiology and clinical outcome of all HCL-patients treated in Innsbruck from 1990 to 2013. Material and Methods: 84 patients were identified and their outcome was evaluated retrospectively. Results:  82.1% were males and the median age was 56 years (range: 25–82). 76 patients (90.5%) received first-line therapy with cladribine being the agent of choice in 43/76 (56.6%) cases. Detailed assessment of response was available in 33/43 (76.7%) patients (OR 100%, CR 84.8%, PR 15.2%). Median PFS was 144 months (95%-CI: 56–232). 27/76 (35.5%) patients received Interferon-α as first-line therapy. Detailed response was evaluable in 63.0% of cases (OR 88.2%, CR 29.4%, PR 58.8%). Median PFS was 30 months (95%-CI: 26–34, n =19). The difference in PFS of patients treated with cladribine and interferon-α was statistically significant (p < 0.001). As third first-line option splenectomy was performed in 7.9% of patients. With a median follow-up of 117 months the relapse rate was 69.7%. 47.4% of patients received cladribine as second-line therapy (OR 100%, CR 78.6%, 21.4%) with a median PFS of 59 months. Further second-line options were interferon-α (39.5%), pentostatin (5.3%),

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Abstracts rituximab (2.6%) and others (5.2%). Median overall-survival has not been reached. Conclusions:  With regard to both epidemiology and clinical outcome the HCL-population treated in Innsbruck is in accordance with the literature. Cladribine was able to achieve complete responses in both treatment-naive and pretreated patients. Overall-survival in this unselected HCL-cohort is excellent.

P06 Proteasome inhibitors and the unfolded protein response in the treatment of multiple myeloma Bojana Borjan1, Gerold Untergasser1, Eberhard Gunsilius1, Johann Kern1,2  epartment of Internal Medicine V, Medical University of D Innsbruck, Innsbruck, Austria 2 Oncotyrol GmbH, Innsbruck, Austria 1

Introduction:  The implementation of proteasome inhibitors (PI) was a big milestone in MM therapy. However, after some time of treatment, patients develop a resistance to these compounds and some patients don’t respond at all to PI. A  molecular pathway, called the unfolded protein response (UPR), induced in stressed and secretory cells, comes more and more in the focus of discussions about reasons for such resistances. Therefore, this study aims to elucidate the role of the UPR in the treatment of MM with PI. Material and Methods:  To cover the genetic heterogeneity of multiple myeloma, various MM cell lines are used in these experiments. These cell lines are treated with bortezomib and carfilzomib, respectively, to measure the activation-level of the UPR, before and after treatment. Cells are also treated with UPR inducers like tunicamycin and thapsigargin to adjust different levels of the UPR to investigate if there is a change in sensi­tivity to PI. Regulated Members of the UPR will be further investigated by using knock-down (shRNA) and knock-out (CRISPR/ Cas9) techniques. Results and Conclusions:  Preliminary data show a corre­ lation between the activation-level of UPR and the response to PI of MM cells. So far, we were able to show that the X-box binding Protein 1 (XBP-1), a prominent member of the UPR, is regulated during the treatment with PI. Therefore, we conclude that the UPR might play an important role in the treatment of MM. Updated data will be presented.

P07 Survival benefit of anti-thymocyte-globulin in hematopoietic stem cell transplantation depends on HLA-C group killer-cell Ig-like receptor (KIR) ligand status Johannes Clausen1, Alexandra Böhm1, Veronika Buxhofer-Ausch1, Sigrid Machherndl-Spandl1, Josef König1, Ansgar Weltermann1, David Nachbaur2 1

 epartment of Internal Medicine 1, Elisabethinen Hospital D Linz, Linz, Austria

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2

 epartment of Internal Medicine V, Medical University of D Innsbruck, Innsbruck, Austria

Introduction:  Anti-thymocyte globuline (ATG) is used to decrease severe acute and chronic graft versus host disease (GVHD) following allogeneic hematopoietic stem cell transplantation (HSCT). However, ATG has not been shown to improve survival in randomized studies. It is unknown whether ATG has different effects in subgroups defined by the recipient’s HLA-C KIR ligand status (C1/1, C1/2, or C2/2). Methods:  We have therefore analyzed retrospectively a twocenter cohort of 720 consecutive, related (n =382) or unrelated (n =338) HSCT, performed between 11/1988 and 10/2014. AntiT-cell serotherapy (predominantly ATG-Fresenius) was applied in 307 transplants. Results:  Following HLA-matched HSCT in C1/1 homozygous patients (n =230), the cumulative incidence of non-relapse mortality (NRM) at 4y was significantly lower following ATGversus non-ATG based transplants (20.3% vs. 39.0%; p =0.004), resulting into a trend for superior 4y-survival in ATG-treated recipients (47.9% vs. 39.6%; p =0.076). No such effect of ATG was found in corresponding C1/2 or C2/2 patients. By multivariate analysis considering established confounding factors, C1/1, but not C1/2 or C2/2 peripheral blood transplant recipients significantly benefited from ATG in terms of less NRM (RR, 0.42; p =0.006), without an increased relapse incidence (RR  1,08; p =0,8), which resulted in reduced overall mortality (RR,  0.60; p =0.046). Conclusions:  In the era of high-resolution HLA-typing, immunological determinants as the HLA-C KIR ligand status may be more important for the decision for or against ATG, than is the donor-recipient family relationship. In recipients belonging to the C1/1 group (comprising nearly 40 % of HSCT recipients), ATG should be considered in peripheral blood HSCT from matched siblings.

P08 A phenotypic comparison of leukemic stem cells in AML, CML and mast cell leukemia Gregor Eisenwort1, 2, Harald Herrmann3, Irina Sadovnik2, Sabine Cerny-Reiterer1, 2, Katharina Blatt2, Thomas Rülicke4, Michael Willmann5, Martin Bilban6, Peter Valent1, 2  udwig Boltzmann Cluster Oncology, Medical University of L Vienna, Vienna, Austria 2 Department of Medicine I, Medical University of Vienna, Vienna, Austria 3 Department of Radiotherapy, Medical University of Vienna, Vienna, Austria 4 Institute of Laboratory Animal Science, University of Veterinary Medicine, Vienna, Austria 5 Department of Companion Animals and Horses, University of Veterinary Medicine, Vienna, Austria 6 Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria 1

The role of leukemic stem cells (LSC) in the development, propagation, and relapse after therapy in patients with AML, CML, and mast cell leukemia (MCL), has become the focus of research in recent years. Notably, substantial effort has been made to develop novel, LSC-eradicating and thus curative treatment approaches in these leukemias. We and others have

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Abstracts shown that the NSG-mouse-engrafting LSC in these leukemias reside in CD34+  subpopulations. In the present study, we analyzed LSC in AML, CML and MCL phenotypically by flow cytometry, qPCR and gene arrays, in order to establish marker- and target expression profiles. We found that LSC in CML express CD25, CD26 and IL-1RAP in an aberrant manner, whereas normal stem cells lack these antigens. Conversely, LSC in AML expressed CD25 and CLL-1, but did not express CD26 and often also lack IL-1RAP. MCL LSC expressed IL-1RAP in a substantial subset of cases, but did not express CD25, CD26, or CLL1. The cell surface targets CD33, CD52, and CD117 (KIT) were detectable on LSC in AML, CML, and MCL. Correspondingly, targeted drugs directed against these surface antigens were found to eliminate LSC in vitro. Together, our findings show that LSC in AML, CML and MCL display a characteristic and unique phenotype and major targets of therapy. A next logical step would be to develop antibody-based LSC-targeting treatment approaches in order to pave the way for curative therapies in these malignancies.

P09 Feasibility of next generation sequencing for routine diagnostic genetic profiling in myeloid malignancies Lukas Gaksch1, Karl Kashofer2, Gerald Höfler2, Christoph Tinchon3, Peter Krippl4, Armin Zebisch1, Hildegard Greinix1, Heinz Sill1, Albert Wölfler1  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Institute of Pathology, Medical University of Graz, Graz, Austria 3 Department of Hemato-Oncology, Hospital Leoben, Leoben, Austria 4 Department of Internal Medicine, Hospital Fürstenfeld, Fürstenfeld, Austria 1

Introduction:  Whole genome and exome sequencing approaches have revealed recurrent mutations in myeloid malignancies with prognostic relevance in terms of response to therapy as well as disease-free and overall survival. The availability of next generation sequencing methods for routine diagnostics makes genetic profiling attractive for incorporation into clinical decision algorithms. Material and Methods: We have established next gener­ ation sequencing using an Ion Torrent Sequencing platform and an amplicon panel covering 19 genes for diagnostic mutational analysis in myeloid neoplasms. Here we report the first results of this diagnostic approach in a clinical setting. Results:  Bone marrow or peripheral blood samples of 55 patients with AML, 5 patients with MDS and two patients with CMML were analysed. In total we identified 146 mutations with a median number of 2 mutations per patient (range 0-6). In 6 cases (9.6%) no mutation was detected. The most frequently mutated genes were NPM1, DNMT3A and TET2. Twenty-seven mutations displayed a decreased allelic frequency indicating the presence of these mutations only in subclones of the disease. Median time from sending in material for analysis to the report of full results was 21 days. Conclusions:  Genetic profiling of diagnostic samples of myeloid malignancies using the Ion Torrent Sequencing platform is clinically feasible and may be useful for exact prognosti-

cation of patients with myeloid malignancies as clinical decision algorithms will increasingly rely on mutational analysis.

P10 RAF kinase inhibitor protein is involved in the myelomonocytic differentiation of hematopoietic progenitors Stefan Hatzl1, Olivia Geiger1, Erika Nußbaumer1, Angelika Rosenberger1, Jakob Troppmair2, Walter Kolch3, Karen Blyth4, Heinz Sill1, Albert Woelfler1, Armin Zebisch1  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Daniel Swarovski Research Laboratory, Medical University of Innsbruck, Innsbruck, Austria 3 The Conway Institute, University College Dublin, Dublin, Ireland 4 The Cancer Research UK Beatson Institute, Glasgow, United Kingdom 1

Introduction:  A somatic loss of RAF kinase inhibitor protein (RKIP) expression is frequently observed in acute myeloid leukemia (AML). As this also correlates with myelomonocytic/ monocytic AML phenotypes, we asked for a potential functional involvement of RKIP in myelomonocytic differentiation. Materials and Methods:  Myelomonocytic differentiation of immature HL-60 AML cells was induced by incubation with 1,25-dihydroxyvitamin D3 (Vit-D3) and measured by the expression of the surface markers CD11c and CD14. The same experiments were performed following modulation of RKIP expression, either by transfecting a GFP-tagged RKIP expression construct or by siRNA knockdown of RKIP, respectively. To  evaluate the role of RKIP in normal hematopoiesis, we assessed GM-CSF induced myelomonocytic differentiation of lineage negative bone marrow (BM) progenitor cells isolated from RKIP knockout mice and compared it to wild-type littermates. Differentiation was assessed by the expression of CD11b, Ly6G and CD115, RKIP protein expression was monitored by Western blot. Results:  Vit-D3 induced myelomonocytic differentiation of immature HL-60 AML cells resulted in a significant downregulation of RKIP expression. Importantly, siRNA knockdown of RKIP increased maturation, whereas its overexpression inhibited this process. The same effects could be shown in murine hematopoietic progenitors, where RKIP depletion was able to increase GM-CSF driven myelomonocytic differentiation. Interestingly, RKIP downregulation in absence of GM-CSF treatment was insufficient to cause maturation, suggesting that it functions as amplifier rather than inducer of this process. Conclusions:  Our data indicate that RKIP is of functional relevance in myelomonocytic differentiation of both, normal and leukemic hematopoiesis.

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Abstracts

P11 Calreticulin in praxi Thomas Jäger1, Denise Aldrian2, David Wanner2, Gerold Untergasser2, Alois H. Lang1, Michael Steurer2 1 2

Hospital Feldkirch, Feldkirch, Austria Medical University of Innsbruck, Innsbruck, Austria

Introduction and Methods: It was recently discovered that somatic mutations in exon 9 of the Calreticulin gene is the second most frequent mutation after JAK2V617F in ET and PMF patients. These CALR mutations were described mutually exclusive with JAK2V617F and MPL mutations. We analysed retrospective data from 70 patients with myeloproliferative disease (MPN) with 71 calreticulin (CALR) mutations, 49 patients were evaluable. We addressed for differences between the most common mutations in calreticulin. Results:  Median age was 66 years, 53% were women. Notably, our data show a higher occurrence of cardiovascular complications in men (12% vs 36%). This was particularly striking in type II CALR mutations (11% vs 50%). The platelet count at diagnosis did not differ between the groups. Moreover, our analyses show that CALR 2 mutations did almost exclusively occur in ET, while CALR 1 mutations were evenly distributed between ET and PMF (CALR 1 mutation: 1 PV, 16 ET, 12 PMF, 2 MPN NOS; CALR 2: 1PV, 10 ET, 0 PMF, 4 MPN NOS, other CALR mutations: 1 ET, 1 MPN NOS, CALR 1+ 2: PMF). A difference in leucocyte count, haemoglobin, gender or total amount of complications was not found between CALR1 and CALR2 mutation. Of note, we found combined JAK2V617F and CALR mutations in 5  patients and 3  other CALR mutations by sequencing. Additionally we found a type I and type II CALR mutation occuring in the same patient. Conclusions:  Our data show a similar clinical presentation at diagnosis for both CALR mutational groups. We found, however, a difference in complications regarding gender. Such a difference is not known in NPMs with JAK2 mutations. Our data support the assumption of different effects of each mutation by unequal occurrence between ET and PMF.

P12 Targeting of BRD4 results in inhibition of proliferation and survival of JAK2 V617F+ MPN cells Alexandra Keller1, Barbara Peter1,2, Johannes Zuber4, Peter Valent1,2, Emir Hadzijusufovic1,2,4  udwig Boltzmann Cluster Oncology, Medical University of L Vienna, Vienna, Austria 2 Division of Hematology and Hemostaseology, Department of Internal Medicine I, Medical University of Vienna, Vienna, Austria 3 Research Institute of Molecular Pathology, Vienna, Austria 4 Department of Companion Animals & Horses, Small Animal Clinic, Internal Medicine, University of Veterinary Medicine Vienna, Vienna, Austria 1

Myeloproliferative neoplasms (MPN), like polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF), are stem cell-derived clonal disorders, often

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accompanied by the JAK2 V617F mutation. So far, the only curative approach is stem cell transplantation. Therefore, current research is focusing on new drug targets. Recently, the epigenetic reader bromodomain-containing protein 4 (BRD4) has emerged as a promising new target in hematological malignancies. We examined the expression and potential value of BRD4 as molecular target in MPN. In our experiments, we employed two JAK2 V617F+  cell lines (SET-2, HEL) as well as primary MPN samples (4 ET patients; 1 PV patient). As assessed by immunocytochemistry, SET-2 and HEL cells were found to express BRD4. As assessed by 3H-thymidine uptake, three different inhibitors of BRD4 (JQ1, BI2536, BI6727; applied for 48 hours) were found to inhibit the proliferation of SET-2, HEL, and primary MPN cells, with the following IC50 values (nM): SET-2: JQ1, 50-100; BI2536, 20-40; BI6727, 50-75; HEL cells: JQ1, 100-500; BI2536, 20-40; BI6727, 30-50; primary MPN cells: 500-1000 with all three drugs. As determined by flow cytometry, BRD4-targeting drugs also induced apoptosis in the two cell lines, with ED50 values of 5 µM for JQ1 and 0.5-5.0 µM for BI2536 and BI6727. Preliminary experiments suggest that JQ1, BI2536 and BI6727 induce a G2/M cell cycle arrest in JAK2 V617F-transformed cells. Together, our data show that MPN cells express BRD4 and that blocking of BRD4 decreases myeloproliferation. The clinical relevance of BRD4-targeting in MPN remains to be determined.

P13 Late remissions following treatment with lenalidomide or thalidomide for mucosa-associated lymphoid tissue lymphoma (MALT lymphoma) – a matter of time? Barbara Kiesewetter1, Marlene Troch2, Werner Dolak3, Leonhard Müllauer4, Markus Raderer1  ivision of Oncology, Department of Medicine I, Medical D University of Vienna, Vienna, Austria 2 Unit for Bone Marrow and Stem Cell Transplantation, Department of Medicine I, Medical University of Vienna, Austria 3 Division of Gastroenterology and Hepatology, Department of Medicine III, Medical University of Vienna, Vienna, Austria 4 Department of Clinical Pathology, Medical University of Vienna, Vienna, Austria 1

Introduction:  Lenalidomide and thalidomide are both substances tested for the treatment of MALT-lymphoma. Particularly lenalidomide appears to be a promising compound; however, long-term results are missing. As late-onset remissions had been registered, we have systemically analyzed patients treated at our institution for long-term improvement. Methods:  Within this retrospective data analysis we could identify 25 patients treated with lenalidomide (n = 18) or thalidomide (n = 7), respectively. All patients were followed according to a standardized follow-up protocol. Results:  28% (7/25) of patients experienced delayed-onset responses without further treatment including four patients (16%) in whom the initial outcome was even switched to a better result (PR to CR n =1, SD to PR n =1, SD to CR n =1, PD to CR n = 1) after a median time of 19.5 months (range; 10.9–32.0), further two patients with ongoing shrinkage of the target lesion for 47.4+ and 43.5+ months, respectively and one patient with durable disease stabilization for 16.2+ months. The median time to best response for all responding patients (13/25, 53%)

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Abstracts was 7.3 months (interquartile range; 6.3-27.3). After a median follow-up of 46 months (interquartile range 32.0–58.5) for the entire collective, 92% (23/25) of patients are alive while one has died due lymphoma progression and one for treatmentunrelated reasons. Conclusions:  Our findings add crucial information to the knowledge of the treatment for refractory/ relapsed MALT-lymphoma with lenalidomide as it appears to be of major importance to assure sufficient follow-up time after treatment with these compounds and before initiation of further therapy as “late-onset responses” may occur.

P15 Hyperglycemia is an adverse risk factor in AML patients undergoing allogeneic hematopoietic stem cell transplantation Sonja Kremser1, Felix Aberer2, Wilma Zinke-Cerwenka1, Peter Neumeister1, Armin Zebisch1, Hildegard Greinix1, Heinz Sill1, Harald Sourij2, Albert Wölfler1  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Division of Endocrinology and Metabolism, Department of Internal Medicine, Medical University of Graz, Graz, Austria 1

P14 Is gender a risk factor for secondary cardiovascular events in R-CHOP treated DLBCL patients? Florian Kocher1,2, Andreas Volgger1, Wolfgang Willenbacher1, Michael Fiegl1, Wolfgang Hilbe1,3  epartment of Internal Medicine V, Medical University of D Innsbruck, Innsbruck, Austria 2 Tyrolean Cancer Research Institute, Innsbruck, Austria 3 Department of Internal Medicine I, Wilhelminenspital Wien, Vienna, Austria 1

Introduction:  Cardiotoxicity is a well-known late effect of anthracycline treatment. Some studies indicate that women are at increased risk to develop cardiac side effects following anthracycline treatment. These results were deduced from studies investigating survivors of childhood cancers. Prompted by these findings we investigated the impact of gender on the development of cardiovascular events in a consecutive cohort of DLBCL patients treated with R-CHOP. Methods and Results:  184 patients diagnosed with DLBCL between 2000 and 2012 at Medical University of Innsbruck were included. Parameters of most interest were pre-existent cardiovascular comorbidities (all grades) and cardiovascular events (heart failure, conduction disorders, acute coronary syndrome, valvular replacement) appearing during treatment or follow-up. Results Mean age at diagnosis was 60.2 in women (n = 99; 53.8%) and 55.9 years in men (n = 85, 46.2%) (p = 0.076), respectively. Both genders were characterized by a comparable burden of cardiovascular comorbidities (women 53.1%, men 46.9%, p = 0.170). The mean administered cumulative dose of anthracycline was similar (women 284 mg/m², men 296 mg/m²; p = 0.383). Overall, 14 women (16.5%) and 9 men (9.1%) developed a secondary cardiac event (p = 0.179). Distribution of different cardiovascular events (p = 0.141) and time to cardiovascular event (5-year EFS; women 82.5%, men 95.6%; p = 0.124) was comparable between the groups. Appearance of a cardiovascular event was not associated with reduced overall survival (p = 0.474). Conclusion:  Our data indicate that cardiovascular events are considerable complications in DLBCL patients. In our series gender did not prove to be a risk factor with respect to cardiotoxicity. Nevertheless, adequately powered prospective studies are needed to validate our results.

Introduction:  Hyperglycemia is a well-defined risk factor for adverse outcome in severely ill patients. However, little is known about its role in the setting of acute myeloid leukemia (AML) and allogeneic hematopoietic stem cell transplantation (SCT). Material and Methods:  We performed a retrospective analysis of 160 AML patients undergoing allogeneic SCT and determined the impact of hyperglycemia (defined as 20% of blood glucose levels higher than 125 mg/dl) within the first 30 days after transplantation on outcome measures. Results:  With a median of 24 blood glucose measurements per patient 57 patients (35.6%) met the criteria for hyperglycemia. During a median follow-up of 10 months 35 of hyperglycemic patients (61.4%) died. In contrast, only 41 out of 103 patients (39.8%) with normal glucose levels succumbed to death (p = 0.007). Median overall survival for patients with hyperglycemia was significantly shorter as compared to patients with normal glucose levels (6.8 months vs. 34.2 months, p < 0.001). Accordingly, estimated overall survival after 5 years was lower in patients with hyperglycemia (30.3% vs. 49.9%) compared to patients with normal blood glucose levels. In multivariate analysis including age at diagnosis, remission status at time of SCT, cytogenetic risk profile, HLA-compatibility and conditioning regimen, hyperglycemia remained an independent risk factor for overall survival. Conclusions:  Hyperglycemia during the first 30 days after allogeneic SCT is an independent risk factor for overall survival in AML patients. Prospective studies testing a strict blood glucose management are warranted in this clinical setting.

P16 Integrin-linked kinase: A devILKin required for chronic lymphocytic leukemia cell proliferation Peter Krenn1,2, Sebastian Hofbauer1,2, Susanne Pucher1,2, Evelyn Hutterer1,2, Elisabeth Hinterseer1,2, Daniela Asslaber1,2, Daniel Neureiter3, Richard Greil1,2, Tanja Hartmann1,2  IMCR, Third Medical Department, Paracelsus Private L Medical University, Salzburg, Austria 2 Salzburg Cancer Research Institute, Salzburg, Austria 3 Institute of Pathology, Paracelsus Private Medical University, Salzburg, Austria 1

Introduction:  Integrin-linked kinase (ILK) is a multifunctional intracellular adaptor protein downstream of beta1 inte-

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Abstracts grins and is involved in tumor cell migration and cell cycle progression. The proliferation of chronic lymphocytic leukemia (CLL) cells requires the lymphoid microenvironment. Here, we studied ILK expression, regulation and function during the crosstalk of CLL cells with this microenvironment. Methods:  ILK expression was cytometrically assessed in peripheral blood (PB) CLL cells. Co-expression of ILK, Ki-67 and the NF-kappaB subunit p65 in CLL lymph nodes (LN) was immunohistochemically analyzed. The LN microenvironment was mimicked in vitro by co-cultures of CLL cells with activated T cells or stimulating CLL cells by IL2/CpG. The role of TNFalpha and NF-kappaB was studied using appropriate inhibitors. Analysis of co-localization of ILK and the centrosomal marker gamma-tubulin was performed by immunofluorescence Results:  We found increased ILK expression in PB CLL cells displaying markers of poor prognosis. Furthermore, recently proliferated CLL cells expressed higher ILK than quiescent cells. Consistently, ILK expression was associated to proliferative lymphoid areas with marked p65 expression. Under co-culture conditions mimicking the activating LN microenvironment, ILK transcription in CLL cells was initiated alongside with cyclin D1 upregulation and followed by translation into protein. This induction was based on NF-kappaB activation by soluble co-culture produced TNFalpha. The newly synthesized ILK protein colocalized to centrosomal structures. Conclusions:  In summary, we describe a TNFalpha-NFkappaB mediated induction of ILK during CLL cell cycle progression within the activating LN microenvironment and propose a role of ILK in organizing centrosomes in proliferating CLL cells.

P17 PDGFRB function and signaling in anaplastic large cell lymphoma Madeleine Kuroll , Nicole Prutsch , Michaela Schlederer2, Tanja Limberger2, Astrid Aufinger1, Suzanne Turner3,5,6, Olaf Merkel1,6, Lukas Kenner1,4,6 1

1

 linical Institute of Pathology, Medical University of Vienna, C Vienna, Austria 2 Ludwig Boltzmann Institute for Cancer Research, Vienna, Austria 3 Department of Pathology, University of Cambridge, Cambridge, United Kingdom 4 Department of Laboratory Animal Pathology, University of Veterinary Medicine, Vienna, Austria 5 Division of Molecular Histopathology, Addenbrooke’s Hospital, Cambridge, United Kingdom 6 European Research Initiative on ALK-related malignancies, http://www.erialcl.net 1

Anaplastic large cell lymphoma (ALCL) is a highly proliferating non-Hodgkin lymphoma of T-cell origin that affects children, young adults and patients over age 60. The nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) fusion protein is identified as the driver of lymphomagenesis in approximately 50 % of ALCL patients. Still the molecular events in ALK-positive and especially ALK-negative ALCL are largely unknown. We want to investigate the role of platelet derived growth factor receptor beta (PDGFRB) in a murine ALCL model as we

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could previously show that the downstream effectors of NPMALK, JUN and JUNB, directly upregulate PDGFRB. Therapeutic inhibition of PDGFRB by imatinib prolonged life in transgenic mice and resulted in sustained remission of a late-stage patient with refractory ALK+ , PDGFR+  ALCL. Moreover, in a retrospective study of 98 fully annotated ALCL patient samples, high PDGFRB expression in tumor or stromal cells correlated with bad prognosis. To elucidate the functional role of PDGFRB, the CD4NPM-ALK transgenic ALCL mouse model was backcrossed to mice with floxed PDGFRB alleles and a CD4-Cre transgene (NPM-ALK/PDGFRBΔT-CELL). NPM-ALK/PDGFRBΔT-CELL mice have a significantly prolonged survival rate suggesting that PDGFRB contributes to tumor development, however is not essential for this process. In addition, the thymic CD4+ / CD8+  T-cell lymphoma phenotype observed in CD4-NPM-ALK transgenic mice becomes CD4-/CD8- in NPM-ALK/PDGFRBΔTCELL mice. These findings indicate that PDGFRB is essential for the progress of T-cells to the double positive stage of T-cell development. We therefore postulate that PDGFRB is crucial for lymphomagenesis by being responsible for accurate T-cell development in the thymus.

P18 The influence of insurance status and travel distance to the treating institution in diffuse large B-cell lymphoma Teresa Magnes1, Thomas Melchardt1, Lukas Weiss1, Clemens Hufnagl1, Stefan Habringer1, Daniel Neureiter2, Wolfgang Tränkenschuh2, Richard Greil1, Alexander Egle1 Department of Internal Medicine III, Cancer Center, LIMCR, Paracelsus Private Medical University, Salzburg, Austria 2 Institute of Pathology, Paracelsus Private Medical University, Salzburg, Austria 1

Introduction:  Despite the knowledge of clinical and molecular risk factors in diffuse large B-cell lymphoma (DLBCL) information on social factors influencing survival is scarce. Earlier studies in patients with different hematologic malignancies and solid tumors suggested that both travel distance to the treating institution and the insurance status influence the clinical outcome. According to our knowledge this is the first study in Europe to analyze the association between insurance status and travel distance and survival in DLBCL. Material and Methods:  Between 2003 and 2014 265 patients with DLBCL were treated with an R-CHOP-like protocol at our tertiary cancer center in Salzburg. Within these patients we evaluated the influence of insurance status and travel distance to the treatment center on the clinical outcome. Results:  The median progression free survival and overall survival for all patients despite their insurance status and the travel distance was 66 months and 94 months, respectively. Despite older age (median 71 vs. 67 years; p = 0.026) of patients with private insurance (54 of 265; 20.4%) there was no difference in the distribution of IPI and NCCN-IPI risk categories compared to patients without private insurance. The median travel distance to the treating institution was 23.8  km (range

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Abstracts 2.3-356 km). Neither for the travel distance nor for the insurance status a significant influence on PFS or OS was found. Conclusions:  This is the first report that clinical outcome of DLBCL patients is not influenced by the insurance status in the Austrian health care system. Furthermore, our data support the safety of centralized lymphoma treatment.

P19 Ultra-deep mass spectrometry-based proteomics enables – new insights into chronic lymphocytic leukemia Rupert Mayer1,2, Astrid Slany1,2, Andrea Bileck1,2, Dominique Kreutz1, Christopher Gerner1,2, Josef Schwarzmeier2  epartment of Analytical Chemistry, Faculty of Chemistry, D University of Vienna, Vienna, Austria Institute for Bioanalytical Oncology, Karl Landsteiner Society, Vienna, Austria

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Introduction:  In spite of extensive research and steadily improving treatment options, CLL still presents an incurable disease with poorly understood pathomechanisms. While genomic approaches have already yielded substantial progress in CLL research, proteomics still remains an incompletely exploited yet powerful tool for the revelation of the molecular background of CLL. Materials and Methods:  B lymphocytes were isolated from blood of healthy donors and CLL patients of three disease progression states. After cell fractionation into cytoplasmic and nuclear proteins, samples were tryptically digested and analysed via ultra-high performance liquid chromatography coupled to high-resolution orbitrap mass spectrometry. Raw data was subjected to MaxQuant software for identification, labelfree quantification and statistical evaluation. Results:  Identification and relative quantification of >  7000 proteins provided us with detailed insights into CLL and normal B-cell proteome. Comparison of all CLL stages against normal B cells resulted in >  1500 significantly regulated proteins. Welldescribed CLL-associated features like up-regulation of ZAP-70 and regulation of ECM-receptor interaction and B-cell receptor signaling were reproduced. Furthermore, many undescribed proteomic regulations were identified like strong increase of PIGR suggesting pronounced tumor-stroma interaction. Contrasting of three different CLL disease states also revealed significant differences in protein expression. Conclusions:  Proteome analysis resulted in a plethora of apparent protein regulation. These rich data allow us to analyze the potential involvement of numerous signaling pathways and relevant molecular players using different strategies. The observed regulation of numerous interesting players on the protein level like PIGR may aid the development of novel therapeutic options for CLL treatment.

P20 Loss of RAF kinase inhibitor protein in acute myeloid leukemia is caused by the overexpressionof miRNA-15a Erika Nußbaumer1, Stefan Hatzl1, Kim Kuepper1, Olivia Geiger1, Rotraud Wieser2, Martin Pichler3, Marcel Scheideler4, Katarzyna Nowek5, Mojca Lavrencic5, Albert Woelfler1, Jakob Troppmair6, Heinz Sill1, Armin Zebisch1  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Department of Medicine I, Medical University of Vienna, Vienna, Austria 3 Division of Oncology, Department of Internal Medicine, Medical University of Graz, Graz, Austria 4 Institute for Diabetes and Cancer, Helmholtz Center Munich, Munich, Germany 5 Department of Hematology, Erasmus University Medical Center, Rotterdam, The Netherlands 6 Daniel Swarovski Research Laboratory, Innsbruck Medical University, Innsbruck, Austria 1

Introduction:  We recently described loss of the tumor- and metastasis-suppressor RAF kinase inhibitor protein (RKIP) as a frequent event in acute myeloid leukemia (AML). As the mechanisms leading to RKIP loss are still unclear, we analyzed the potential involvement of miRNAs within this process. Material and Methods: RKIP protein expression was assessed in 33 AML specimens, which had been characterized by miRNA-ChiP profiling previously. ChiP data of miRNAs related to RKIP expression were further validated by qPCR. To evaluate their potential functional involvement, RKIP expression was monitored by qPCR and Western blot in NB-4 AML cells following transfection with miRNA mimics. Finally, we analyzed microarray and miRNA-ChiP data in an independent cohort comprising 214 AML patients. Results:  By combining RKIP expression and miRNA-ChiP data, we identified a set of eight miRNAs with aberrant expression in AML specimens with RKIP loss. Increased expression of miRNA-23a (P = 0.019), miRNA-24 (P = 0.043) and miRNA-15a (P = 0.043) could be validated by means of qPCR. Importantly, in experiments using miRNA mimics, overexpression of miRNA15a markedly decreased RKIP mRNA and protein expression, indicating that it is functionally involved in the downregulation of this tumor-suppressor. Finally, analysis of an independent AML cohort comprising more than 200 patients further corroborated its association to decreased RKIP expression, thereby supporting the relevance of this finding. Conclusions:  Our data demonstrate that overexpression of miRNA-15a is associated with the downregulation of RKIP in AML and functionally involved in this process.

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P21 A retrospective evaluation of negative effects of rituximab maintenance therapy in patientswith Non-Hodgkin’s Lymphoma Nadine Olah, Felix Keil Division of Hematology and Oncology, Hanusch Hospital, Vienna, Austria Introduction:  Rituximab (RTX) maintenance has been proven an effective treatment for patients with NHL after receiving chemotherapy. It showed significant elongation of the progression-free survival in patients with follicular lymphoma, thus had no influence on overall survival. However, side effects of this long-term therapy are unclear. Methods:  35 patients receiving a maintenance therapy after a RTX-including protocol were included. Most patients were diagnosed with follicular lymphoma FL (N = 21) or mantle-cell lymphoma MCL (N = 10). 4 patients suffering form CLL were included as part of the MABTENANCE study. In most patients the applied dose was 375 mg/m2 every 2 or 3 months for 2 years. Results:  35 patients were identified within a period from 2007-2015, 12 are still undergoing therapy. The maintenance therapy was applied for a median number of 8 cycles, ranging from 3 to 12. In 5 patients the therapy had to be terminated prematurely (p = 0.14), either of prolonged neutropenia (N = 4) or relapse (N = 1) within a median of 7 cycles (range 3-8). Respiratory (N = 12, p = 0.14) and also urinary tract infections (N = 2, p = 0.06) were found in 12 patients within a median of 7 cycles. In 21 % an immunoglobulin deficiency was detected. Conclusions:  It could be found, that in 35% of all patients undergoing maintenance therapy infections and side effects like elongated neutropenia occurred. A significant correlation in patients with antibody deficiency and the occurrence of infections could be verified (p = 0.219).

P22 Treatment of high risk aggressive lymphomas with DA EPOCH R – a single center retrospective analysis Michael Panny, Thomas Nösslinger, Michaela Möstl, Elisabeth Menschel, Theresia Kornberger, Ralph Simanek, Carlos Andres Corena Herrera, Gregor Nemschak, Nadine Olah, Felix Keil Deparment of Internal Medicine 3 (Hematology and Oncology), Hanusch Hospital, Vienna, Austria Introduction:  Promising results in patients suffering from high risk DLBCL, Burkitt`s lymphoma (BL), mediastinal grayzone lymphoma (MGZL) and primary mediastinal B cell lymphoma (PMBCL) treated with DA-EPOCH R have been reported. In our center high risk DLBCL – defined as double-hit/double hit score 2 or high risk IPI – BL, MGZL and PMCL are treated with DA EPOCH R. Methods:  Retrospective analysis of toxicity and efficacy in DA EPOCH R treated patients. So far 11 previously untreated patients – 8 male, 3 female – with a median age of 60 a (32 a – 75 a)

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have been treated with a total of 48 cycles of DA EPOCH R: 7 DLBCL, 1 MGZL, 1 PMBCL, 2BL. Results:  Grade III/IV ANC occurred in 71 % of all cycles, thrombocytopenia III/IV° in 17%, grade III anemia in 4% of 48 cycles. Dose escalation was possible in 10 cycles (21%) – in none of patients aged >  65 a dose escalation was possible. Due to peripheral sensory neuropathy II-III° in 6 patients Vincristine had to be dose reduced in 31% of all cycles. Other CTCAE grade III non-hematopoietic toxicities were: peripheral motor neuropathy, mucositis, colitis, hyperglycemia – each in 1 patient. So far 7 patients finished treatment: 3 in CR, 1 in PR, 1 relapsed early, 1 had treatment failure, 1 patient had to be switched to a less toxic regimen due to repeated febrile neutropenia. Conclusions  Although limited data DA EPOCH R seems to be a feasible treatment with acceptable toxicity. Only in young rather fit patients dose escalation seems to be possible.

P23 Stammzellmobilisierung mit Plerixafor bei Familienspendern für allogene, TCRαβ/CD19 depletierte periphere hämatopoietische Stammzelltransplantation bei Kindern Thomas Perwein, Herwig Lackner, Wolfgang Schwinger, Andrea Raicht, Daniela Sperl, Petra Sovinz, Christian Urban Abteilung für Pädiatrische Hämato-Onkologie, Universitätsklinik für Kinder- und Jugendheilkunde, Medizinische Universität Graz, Graz, Österreich Einleitung:  Im Vergleich zur konventionellen Stammzellmobilisierung mit G-CSF im Rahmen der autologen und allogenen peripheren Stammzelltransplantation (PSZT) führt die zusätzliche Verabreichung des kompetitiven SDF-1α-CXCR4– Antagonisten Plerixafor zu einer besseren Stammzellmobili­ sierung und höheren Stammzellausbeute. Methoden:  15 Kinder wurden von 2011 bis 2014 an unserer Abteilung einer allogenen, kombiniert TCRαβ/CD19 depletierten und CD34+  selektionierten PSZT von FamilienspenderInnen (6 HLA-idente und 9 haploidente) unterzogen. Die Stammzellmobilisierung erfolgte mittels Filgastrim 10 μg/ kg subkutan täglich, mit Apheresestart am Tag 5 (Gruppe I, n = 7). Gruppe II (n = 8) erhielt zusätzlich Plerixafor 0,24 mg/kg subkutan 11 Stunden vor Apherese. Ergebnisse:  Bei Apherese war die Zahl der zirkulierenden Stammzellen in Gruppe II signifikant höher als in Gruppe I (Median 181,5/μl [78-291] vs. 65,5/μl [47-127]; p = 0,008), was auch in einer höheren Stammzellausbeute in Gruppe II resultierte (Median 24,7 x 10e6 [8,3-65] vs. 8 x 10e6 [2,1-65,1] CD34+ Zellen/kg Spendergewicht; p = 0,04). Die erste Gabe von Plerixafor bewirkte einen fünffachen Anstieg der CD34+  Zellen im Blut (p = 0,0007). Bei vergleichbaren Nebenwirkungsraten benötigten 25% der SpenderInnen in Gruppe II nur eine Apherese. Die Gesamtausbeute und Anzahl der transplantierten Stammzellen (Median 22,4 x 10e6 [10,8-30,1] vs. 11,8 x 10e6 [4,82-52,2] CD34+ Zellen/kg) schien in Gruppe II höher, jedoch ohne signifikanten Unterschied (p = 0,46). Das leukozytäre Engraftment trat in beiden Gruppen im Median am Tag 10 ein, trotz myeloablativer Konditionierung in Gruppe II. Ein stabiles Engraftment wurde bei 71,4% in Gruppe I vs. 87,5% in Gruppe II erreicht. Schlussfolgerungen:  Die Kosten von Plerixafor rechtfertigen sich durch ein schnelles und stabiles Engraftment sowie

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Abstracts durch eine höhere Stammzellausbeute bei erster Apherese, mit Reduktion der Mobilisierungsdauer und Apheresetage.

P24 High expression of CXCR4 and CXCR7 is associated with an unfavourable clinical course Beata Pursche1, Kertin Wenzl1, Katharina Troppan1, Martin Pichler2, Christine Beham-Schmid3, Peter Neumeister1, Alexander Deutsch1  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Division of Oncology, Department of Internal Medicine, Medical University of Graz, Graz, Austria 3 Institute for Pathology, Medical University of Graz, Graz, Austria 1

Introduction:  The two chemokine receptors CXCR4 and CXCR7 were identified to be implicated in multiple key processes in tumour cells, including proliferation, survival, migration, invasion and metastasis. Because of the limited knowledge on CXCR4 and CXCR7 in diffuse large B cell lymphomas (DLBCL), we aimed to determine the mRNA expression and genetic alteration of CXC4 and CXCR7. Methods:  Therefore, we performed expression analysis by RQ-PCR in 79 DLBCL specimens and direct sequencing in 25  DLBCL specimens with an advanced clinical stage (stage II-IV) for CXCR4 and CXCR7. Results:  Comparing expression levels of both chemokine receptors of DLBCL patients with clinical stage I to patients with an advanced stage, a five-fold higher CXCR4 (p = 0.017) and a 3.5-fold higher CXCR7 (p = 0.087) expression were detected in advanced stage lymphoma specimens. CXCR4 expression positively correlated with bone marrow infiltration (Spearmanrho = 0.550 and p < 0.001) and Kaplan-Meier analyses revealed that a high CXCR7 expression was associated with poor overall survival (p = 0.033). Three of 25 DLBCL specimens exhibited a synonymous mutation (c.414 C> T) in the CXCR4. In CXCR7 locus a silent variation (c.796 C> T) was detected in six of 25 DLBCL specimens, and four of the six mutated specimens exhibited additionally a second silent variation (c.189 C> T). Conclusions:  Our data indicate that CXCR4 and CXCR7 expression are associated with an advanced stage in aggressive lymphomas and clinical outcome parameters. Therefore, these CXCR receptors might serve as useful clinical prognostic marker.

P25 Non-Hodgkin Lymphoma in childhood versus adolescence: clinicopathologic differences and outcome. A population based analysis of the Austrian NHL-BFM group Bettina Reismüller1, Andishe Attarbaschi1, Michael Dworzak1,2, Oskar A. Haas1,2, Christian Urban3, Bernhard Meister4, Ingrid Simonitsch-Klupp5, Helmut Gadner2, Nora Mühlegger2, Georg Mann1 1

 aediatric Haematology and Oncology, St Anna Children’s P Hospital, Vienna, Austria

Children’s Cancer Research Institute, Vienna, Austria Paediatric Haematology and Oncology, Medical University of Graz, Graz, Austria 4 Department of Paediatrics, Medical University of Innsbruck, Innsbruck, Austria 5 Clinical Institute of Pathology, Medical University of Vienna, Vienna, Austria 2 3

Introduction:  The majority of Non Hodgkin Lymphomas (NHL) in childhood and adolescence is highly malignant. Standardized diagnosis, registration, and treatment according to common protocols comprise virtually 100 % of patients diagnosed in Austria. Higher age as a prognostic factor has been found to be significantly correlated with a worse outcome in paediatric studies for acute lymphoblastic leukaemia. Methods:  To address this issue for NHL, we analysed the data collected prospectively of patients (n = 318) treated in Austria in common NHL-BFM protocols between March 1996 and December 2013. Type of NHL was mature B-cell origin in 208  patients, 14 precursor-B lymphoblastic lymphomas (B-LBL), 50  precursor T-lymphoblastic lymphomas (T-LBL), and 46 anaplastic large cell lymphomas (ALCL). At the time of diagnosis, 245 patients were <  15 years and 73 were ≥  15 years old. The male:female ratio was 2.8 : 1. Results:  We compared the distribution of diagnoses, gender, stage, and outcome according to age >  15 versus age <  15 years. Types of NHL, gender, and stage of disease were evenly distributed among the two age groups. However, as for subtypes of mature B-cell neoplasms, Burkitt lymphoma/leukemia occurred more often in younger patients (122/159 [76.7%] <  15 y vs. 11/49 [22.4%] ≥  15 y), whereas diffuse large B-cell lymphoma (DLBCL) was more common in adolescents (25/159 [15.8%] <  15 y vs. 28/49 [57.1%] ≥  15 y). There was no significant difference in probability of eventfree survival at 10 years (10-year pEFS; 82.4% for pts. < 15 y, 71.9% for pts. ≥ 15 y; P = 0.08). We observed a total number of 40 relapses, 27 in the age group < 15 (11.0 %/pts.), 13 were ≥ 15 years old (17.8%/pts). Thirty-five patients died, 24/245 (9.8%) < 15 y, 11/73 (15.7%) ≥ 15 y. There were 3/24 and 3/11 toxic deaths in the two groups, respectively. Conclusions:  We conclude that although age at diagnosis did not significantly influence outcome of NHL in our population-based pediatric cohort, results seem to be better in the younger age group. Our study represents a basis for future investigations comparing pediatric and adult therapeutic approaches in the treatment of NHL in adolescents and young adults.

P26 Updated report of the Austrian CML registry Stefan Schmidt1, Albert Wölfler2, Richard Greil3, Sonja Burgstaller4, Ernst Schlögl5, Andreas Petzer6, Alois Lang7, Ansgar Weltermann8, Daniela Voskova9, Manfred Mitterer10, Peter Valent11, Nicole Eberhard12, Alois Walder13, Klaus Geissler14, Johannes Andel15, Christa Häusler16, Christof Ludescher17, Horst Oexle18, Markus Korger19, Michael Schnallinger20, Stephan Schreieck21, Peter Krippl22, Michael Pober23, Ewald Woell24, Dietmar Geissler25, Ursula Rochau26,27, Uwe Siebert26,27, Josef Thaler4, Günther Gastl1  epartment of Internal Medicine V, Medical University of D Innsbruck, Innsbruck, Austria

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Abstracts  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz 3 Department of Internal Medicine III, Paracelsus Private Medical University, Salzburg, Austria 4 Division of Internal Medicine IV, Klinikum Wels-Grieskirchen, Wels, Austria 5 Department of Internal Medicine III, Hanusch Hospital, Vienna, Austria 6 Department of Internal Medicine I, Hospital Barmherzige Schwestern Linz, Linz, Austria 7 Department of Internal Medicine, Hospital Feldkirch, Feldkirch, Austria 8 Department of Internal Medicine I, Elisabethinen Hospital, Linz, Austria 9 General Hospital Linz, Linz, Austria 10 Hospital Meran, Merano, Italy 11 Division of Hematology and Hemostaseology, Department of Medicine I, Medical University of Vienna, Vienna, Austria 12 Hospital Leoben, Leoben, Austria 13 Hospital Lienz, Lienz, Austria 14 Hospital Wien-Hietzing, Vienna, Austria 15 Hospital Steyr, Steyr, Austria 16 Hospital Dornbirn, Dornbirn, Austria 17 Ambulatorium for Hematology und Onkology, Innsbruck, Austria 18 Hospital Hall, Hall, Austria 19 Hospital Barmherzige Brüder, Eisenstadt, Austria 20 Hospital St. Johann, St. Johann, Austria 21 Hospital Reutte, Reutte, Austria 22 Hospital Fürstenfeld, Fürstenfeld, Austria 23 Klinikum St. Pölten, St. Pölten, Austria 24 Hospital St. Vinzenz, Zams, Austria 25 Hospital Klagenfurt, Klagenfurt, Austria 26 UMIT – University for Health Sciences, Medical Informatics and Technology, Hall, Austria 27 ONCOTYROL – Center for Personalized Cancer Medicine, Innsbruck, Austria 2

Introduction:  Tyrosin kinase inhibitors (TKI) treatment in CML is directed by response kinetics. While the initial ELN recommendations guided treatment according to the achieved cytogenetic response development of standardized BCR-ABL transcript monitoring lead to more stringent treatment goals. Second generation TKIs (2G-TKIs) have demonstrated superior efficacy in inducing deep molecular response, however, in clinical practice the majority of patients are still receiving imatinib. Methods:  The Austrian CML registry is an ethics committee approved database under the patronage of the ASHO. It is focused on diagnostic parameters e.g. conventional cytogenetics and/or FISH, BCR-ABL transcript levels, blood counts, general laboratory parameters, CML phase, concomitant diseases, CML specific treatment and including stem cell transplantation and adverse effects. Results:  Here we summarize the CML registry data as of January 2015 and focus on treatment response. The CML registry cohort comprised a total of 445 patients. A total of 314 evaluable patients were treated at least once with imatinib and 54  and 36 patients were received at least once nilotinib and dasatinib, respectively. Here we will present the results of the currently running final update on the registry cohort. Conclusions:  The majority of patients fulfils the ELN 2013 criteria for optimal response. Imatinib has been for years the mainstay of treatment and is still so. However, the use of 2G-TKIs has increased. But the majority of patients is switched to 2G-TKIs upon failure or intolerance to imatinib. The main focus of this registry for 2015 will be the percentage of patients reaching a sustained deep molecular response.

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P27 Hydroxyurea supresses the growth of CML sub-clones expressing BCR/ABL-T315Iin heavily pretreated patients: retrospective analysis of four cases Mathias Schneeweiss1, Susanne Herndlhofer1, Thomas Lion2, Peter Valent1,3, Karoline Gleixner1  ivision of Hematology and Hemostaseology, Department of D Medicine I, Medical University of Vienna, Vienna, Austria 2 Children’s Cancer Research Institute (CCRI), Vienna, Austria 3 Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria 1

In chronic myeloid leukemia (CML), the occurrence of BCR/ ABLT315I leads to resistance against most tyrosine kinase inhibitors (TKI) and represents a clinical challenge. Ponatinib, which supresses BCR/ABLT315I, cannot be used in all patients because of severe side effects. Thus, other therapeutic strategies must be considered. Hydroxyurea (HU) interferes with DNA-synthesis and is used since many decades for the treatment of CML. We report on four male patients with advanced CML (age 39-65 years) in whom a BCR/ABLT315I positive sub-clone could be supressed or even disappeared during HU-treatment. All patients had been treated with one or more TKI before BCR/ABLT315I was detected. Thereafter, HU was prescribed in an attempt to control CML expansion. During therapy with HU, leukocyte counts and total BCR/ABL remained stable during the observation period (3–12 months). Surprisingly, in 3/4 patients, the sub-clone expressing BCR/ABLT315I disappeared during HU-treatment, and in one patient, the percentage of BCR/ABLT315I, compared to total BCR/ABL, assessed by mutation-specific (ligase-dependent) qPCR, decreased from 94% to 7%. Two of the 4 patients underwent SCT, one of these patients is still alive. In the two other patients, the disease remained stable for 6 and 12 months, respectively. Together, we provide evidence that HU supresses CML sub-clones harbouring BCR/ABLT315I. This observation may have clinical implications for heavily pretreated patients, including those who are prepared for bridging to SCT or who cannot tolerate ponatinib because of safety issues or side effects. HU effects in BCR/ABLT315I+  CML should be further evaluated in controlled clinical trials.

P28 Engraftment nach CD3 TCRαβ/CD19 depletierter hämatopoietischer Stammzelltransplantation bei Kindern und Jugendlichen Daniela Sperl1, Wolfgang Schwinger1, Christian E. Urban1, Roland Wilfing1, Peter Lang2, Markus Seidel1, Volker Strenger1, Petra Sovinz1, Herwig Lackner1, Rupert Handgretinger2  bteilung für pädiatrische Hämatologie/Onkologie, A Medizinische Universität Graz, Graz, Österreich 2 Abteilung für Allgemeine Pädiatrie, Hämatologie/Onkologie, Universitätsklinikum Tübingen, Tübingen, Deutschland 1

Einleitung:  Kinder und Jugendliche mit Hochrisiko-Leukämien, malignen metastasierten Tumoren oder angebore-

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Abstracts nen Stoffwechselerkrankungen können unter bestimmten Voraussetzungen durch eine allogene hämatopoietische Stammzelltransplantation (SCTX) geheilt werden. Die Transplantationsassoziierte Mortalität war in der Vergangenheit v.a. durch die Graft-versus-Host-Disease (GvHD) bedingt sehr hoch. Die Methode der CD3 TCRαβ/CD19 depletierten SCTX ermöglicht die für die GvHD hauptverantwortlichen CD3 TCRαβ+  Zellen aus dem Apharesat zu entfernen und die das Engraftment erleichternden γδ+  T-Lymphozyten und NKZellen zu behalten. Fragestellung:  Kann man bei SCTX durch vorangehende selektive Depletion von TCR-αβ+ Stammzellen das GVHD Risiko der Patienten vermindern ohne das Engraftment zu beeinträchtigen? Methoden:  13 Kinder, 7 mit nicht malignen und 6 mit malignen Grunderkrankungen, erhielten eine HLA matched, mismatched oder eine haploidentische SCTX nach ex vivo Entfernung von CD3 TCRαβ+  T-Zellen und CD19+  B-Zellen. Ergebnisse:  Die mediane Anzahl der verabreichten CD34+ , αβ+  CD3+  und B-Zellen war 21,7 x 106/kg, 11,79 x 103/kg und 73 x 103/kg Körpergewicht. Alle Patienten bis auf eine Patientin zeigten ein frühes komplettes 3-Linien-Engraftment, 2 Patienten präsentierten eine auf die Haut beschränkte akute GvHD Grad I-II. Keiner der Patienten entwickelte eine viszerale akute oder chronische GvHD. Die Inzidenz der Transplantationsassoziierten Mortalität war 7,6% (1/13). Die Gesamtmortalität bedingt durch die Grunderkrankung war 50% (6/13). Mit einem medianen Follow-up von 10 Monaten leben 6/13 Patienten und sind ohne Zeichen ihrer Grunderkrankung. Schlussfolgerungen:  Unsere Ergebnisse zeigen, dass diese Methode der Graft-Manipulation geeignet ist, eine GvHD effizient zu verhindern und ein sicheres Engraftment zu gewährleisten.

P29 Distinct chemokine receptor profiles in multiple myeloma cells compared to normalbone marrow cells Kerstin Wenzl1, Beata Pursche1, Daniela Klescher1, Christine Beham-Schmid2, Alexander Deutsch1, Peter Neumeister1, Katharina Troppan1  ivision of Hematology, Department of Internal Medicine, D Medical University Graz, Graz, Austria 2 Institute for Pathology, Medical University Graz, Graz, Austria 1

Introduction:  Multiple Myeloma (MM) is a malignant hematological tumor, characterized by clonal proliferation of malignant plasma cells in the bone marrow (BM). MM is commonly preceded by a common benign plasma cell tumor called Monoclonal Gammopathy of Undetermined Significance (MGUS). Recently, chemokine receptors (CCR) and their ligands have been identified to play a crucial role in normal B-cell differentiation and development of hematopoietic malignancies. Due to the lack of knowledge on the CCRs’ role in MM, we aimed to investigate their expression profile in MGUS and MM patients. Material and Methods: Therefore, we investigated the mRNA expression levels of 18 known chemokine receptors (CCR1-CCR9, CXCR1-CXCR7, XCR1, CX3CR1) by using semi-

quantitative real time PCR on 18 samples of MM and 3 MGUS samples. Five samples of normal BM and 4 plasma cell (PC) samples served as non-neoplastic controls. Results:  Comparing BM samples with MM samples we could detected a de-novo expression of 3 CCRs, namely CCR1, CCR4, and XCR1. Furthermore we could observe a difference in the chemokine receptor profile of MM samples compared to PC. Therein, CCR1, CCR2, CXCR1 and CXCR2 showed a de novo expression in MM samples. Additionally, CXCR4 was upregulated in MM, whereas CXCR5 showed a down-regulation. Conclusions:  Our data indicate that the chemokine receptor expression profile of MM differs substantially from those of non-neoplastic BM and PC. Hence, these multiple deregulated CCRs might serve as useful prognostic tool and might be a valuable clinical marker.

P30 Epsilon-gamma-delta-beta° thalassemia Austrian I: identification of a novel hemoglobinopathy Armin Zebisch1, Eduard Schulz1, Stefan Hatzl1, Michela Grosso2,3, Barbara Lombardo2,3, Giovanni Acierno2,3, Heinz Sill1, Achille Iolascon2,3  ivision of Hematology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Department of Molecular Medicine, University Federico II of Naples, Naples, Italy 3 CEINGE- Advanced Biotechnologies, Naples, Italy 1

Introduction:  We present an Austrian family with inherited microcytic anemia, which could not be diagnosed by conventional laboratory diagnostic approaches. Therefore, we aimed to elucidate its genetic background employing molecular approaches, including next generation sequencing (NGS). Patients and Methods:  The index patient was a 47-year-old Austrian male, presenting with microcytic anemia, reticulocytosis and laboratory signs of hemolysis. Two of his three children demonstrated similar laboratory values and peripheral blood films showed the abundance of target cells, thereby pinpointing towards a hemoglobinopathy. Nevertheless, conventional hemoglobin electrophoresis revealed no signs of thalassemia and sequencing of alpha-thalassemia hot-spot variants was also negative. To screen for a novel genetic trait, we performed whole exome sequencing (WES), multiplex ligation-dependent probe amplification (MLPA) and high-resolution array comparative genomic hybridization (hr-CGH). Results:  We were able to diagnose this case as epsilongamma-delta-beta° thalassemia by combination of MLPA and hr-CGH. The causative deletion spanned 339225 bp on the short arm of chromosome 11 and affected the entire cluster of beta globin genes. It has not been described previously and is therefore named Austrian I. Of note, although NGS produced several putative candidate genes, it failed to delineate the proper diagnosis, which highlights potential limitations of this technique. Conclusions:  We identified a novel variant of epsilongamma-delta-beta° thalassemia, named Austrian I. Most importantly, this diagnosis could only be established by combining a detailed clinical characterization with pedigree analysis and a broad range of molecular techniques.

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P31 Aussagekraft des Chen-Scores und Prävalenz von Hochrisikosituationen bei Patienten, die wegen eines Kolorektalkarzinoms FOLFIRI bzw. XELIRI erhalten Temeida Alendar , Elmir Ljubuncic , Klaus Geissler 1

 omprehensive Cancer Center, Medical University of C Vienna, Vienna, Austria 2 Division of Oncology, Department of Medicine I, Medical University of Vienna, Vienna, Austria 3 Department of Pathology, Medical University of Vienna, Vienna, Austria 4 Department of Surgery, Medical University of Vienna, Vienna, Austria 5 Departement of Radiotherapy, Medical University of Vienna, Vienna, Austria 1

Poster Onkologie

1

2

 udwig Boltzmann Institut für klinische Onkologie, Wien, L Österreich 2 5. Medizinische Abteilung mit Onkologie und Palliativstation, Krankenhaus Hietzing, Wien, Österreich 1

Einleitung und Fragestellung: Die Vermeidung neutropenischer Komplikationen ist in der Onkologie für den Erfolg einer Chemotherapie ausschlaggebend. Kombinationstherapien mit 5-FU und Irinotecan haben ein 10-20% Risiko einer febrilen Neutropenie (FN) (Aapro M et al, EJC 2010). Vor kurzem wurde ein hämatologischer Score publiziert (Chen K et al. PLOS one 2014), mit dem Patienten, die eine G-CSF-Prophylaxe benötigen, identifiziert werden können. In unserer Studie wollten wir die Prävalenz von Hochrisikosituationen (ANC < 4,4G/L +  ALC < 2,1G/L +  AMC < 0,28G/L) und die Aussagekraft dieses Scores bei behandlungsbedürftigen Patienten mit Kolorektalkarzinom in einer Real-Life-Situation ermitteln. Material und Methoden:  Bei 36 Patienten, die wegen eines fortgeschrittenen Kolorektalkarzinoms an unserer Abteilung FOLFIRI oder XELIRI erhielten, wurde der Chen-Score retrospektiv vor Chemotherapie errechnet und mit einer allfälligen Aufnahme wegen FN (Fieber > 38° + ANC < 0,5G/L) unter Chemotherapie korreliert. Ergebnisse:  10 Frauen und 26 Männer wurden an unserer Abteilung mit FOLFIRI oder XELIRI behandelt. Das mediane Alter dieser Patientengruppe betrug 66 Jahre (48–81). Eine Hochrisikosituation nach dem Chen-Score wurde in 3 Fällen (8%) festgestellt. Keiner der 36 Patienten wurde im Verlauf der Chemotherapie wegen einer febrilen Neutropenie aufgenommen. Ein Patient der Hochrisikogruppe wurde wegen Fieber > 38° bei einem ANC Wert von 0,77G/L stationär aufgenommen. Schlussfolgerungen:  Unsere Daten weisen darauf hin, dass bei Patienten mit Kolorektalkarzinom, bei denen eine Irinotecan-haltige Kombinationstherapie mit 5-FU geplant ist, nur selten eine Hochrisikosituation (8 %) entsprechend dem Chen-Score vorliegt. Ein normaler Chen-Score scheint in der Real-Life-Situation ein verlässliches und altersunabhängiges Kriterium gegen eine allfällige Wachstumsfaktor Prophylaxe darzustellen.

P32 Prognostic impact of breast cancer (BC) subtype in elderly patients

Background:  We aimed to analyze the impact of BC subtypes on the clinical course with special emphasis on the occurrence of brain metastases (BM) and outcome in an elderly breast cancer population. Patients and Methods: 571 patients ≥ 65 years receiving treatment for BC from 2007-2011 were identified from a BC database. BC subtypes and clinical characteristics including overall survival (OS) were obtained by chart review. Statistical analysis was performed using the Chi Square test, the log rank test and time depended covariate cox regression model as appropriate. Results:  Three-hundred-eighty/571 (63%) were grouped among the young-old (65-74 years), 182/571 (31.9%) among the old-old (75-84 years), and 29/571 (5.1%) among the oldest-old (≥ 85 years). 392/571 (68.8%) patients presented with luminal BC, 119/571 (20.8%) with HER2 positive and 59/571 (10.3%) with triple negative BC. After a median follow up of 38 months (range 0-204), 115/571 (20.1%) patients presented with metastatic recurrence. Highest recurrence rate was observed in HER2 positive BC patients (43/119 (36.1%)), followed by triple negative (15/59 (25.4%) and luminal BC (57/392 (14.5%); p < 0.001; Chi Square test). BM occurred significantly more frequently in HER2 positive BC patients (9/119 (7.6%) compared to triple negative (2759 (3.4%) and luminal BC patients (6/392 (1.5%); p = 0.003; Chi Square test). Occurrence of metastases (HR 7.7; 95% CI: 5.2–11.4; p < 0.001) as well as development of BM (HR 3.5; 95% CI: 1.9–6.4; p < 0.001) had a significant impact on OS prognosis as entered in a time depended covariate cox regression model. Conclusions:  In contrast to younger BC patients, HER2 posi­tive BC subtype and not triple negative BC subtype was linked to the most aggressive clinical course including the development of metastatic disease and BM in our elderly cohort.

P33 Density of tumor-infiltrating lymphocytes correlates with extent of brain edema and overall survival time in patients with brain metastases Anna Sophie Berghoff1,2, Monika Hackl3, Georg Widhalm2,4, Karin Dieckmann2,5, Rupert Bartsch1,2, Harald Heinzl6, Christoph Zielinski1,2, Peter Birner7, Jerome Galon8, Matthias Preusser1,2  epartment for Medicine I, Medical University of Vienna, D Vienna, Austria 2 Comprehensive Cancer Center – CNS Tumors Unit, Vienna, Austria 3 Austrian National Cancer Registry, Vienna, Austria 4 Department of Neurosurgery, Vienna, Austria 1

Elisabeth Bergen1,2, Cornelia Tichy1,2, Anna S. Berghoff1,2, Margaretha Rudas1,3, Robert M. Mader1,2, Karin Dieckmann1,5, Christoph C. Zielinski1,2, Guenther G. Steger1,2, Matthias Preusser1,2, Rupert Barsch1,2

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Abstracts Department of Radiotherapy, Vienna, Austria Center for Medical Statistics, Informatics, and Intelligent Systems, Vienna, Austria 7 Institute of Clinical Pathology, Medical University of Vienna, Vienna, Austria 8 INSERM, UMRS1138, Laboratory of Integrative Cancer Immunology, Paris, France 5 6

Introduction:  The immune microenvironment of the brain differs from that of other organs and the role of tumor-infiltrating lymphocytes (TILs) in brain metastases (BM), one of the most common and devastating complication of cancer, is unclear. Methods:  We investigated TIL subsets and their prognostic impact in 116 BM specimens using immunohistochemistry for CD3, CD8, CD45RO, FOXP3 and PD1. The Immunoscore was calculated as published previously. Results:  Overall, we found TIL infiltration in 115/116 (99.1%) BM specimens and TIL density was not associated with corticosteroid administration (p > 0.05). A significant difference in infiltration density according to TIL subtype was present (p < 0.001; Chi Square); thereby high infiltration was most frequently observed for CD3+ TILs (95/116; 81.9%) and least frequently for PD1+ TILs (18/116; 15.5%; p < 0.001). Highest TIL density was observed in melanoma, followed by renal cell cancer and lung cancer BM (p < 0.001). The density of CD8+ TILs correlated positively with the extent of peritumoral edema seen on pre-operative magnetic resonance imaging (p = 0.031). The density of CD3+ (15 vs. 6 months; p = 0.015), CD8+ (15 vs. 11 months; p = 0.030) and CD45RO+ TILs (18 vs. 8 months; p = 0.006) showed a positive correlation with favorable median OS times. Immunoscore showed a strong and significant correlation with survival prognosis (27 vs. 10 months; p < 0.001) and the prognostic impact of Immunoscore was independent from established prognostic parameters at multivariable analysis (HR 0.612, p < 0.001). Conclusions:  Dense TILs infiltrates are common in BM and correlate with the amount of peritumoral brain edema and with survival prognosis, thus identifying the immune system as promising biomarker for cancer patients with CNS affection.

P34 The pre-operative AST/ALT (De-Ritis) ratio represents a poor prognostic factor in a cohort of non-metastatic renal cell carcinoma patients Angelika Bezan1, Edvin Mrsic1, Daniel Krieger2, Tatjana Stojakovic3, Karl Pummer2, Richard Zigeuner2, Georg C. Hutterer2, Martin Pichler1  ivision of Oncology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Department of Urology, Medical University of Graz, Graz, Austria 3 Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria 1

Introduction:  Aminotransaminases are strongly involved in cellular metabolism, as well as the turnover of cancer cells and represent easily measureable potential blood-based biomarkers. The objective of the current study was to evaluate the prognostic value of pre-operatively assessed Aspartate Amino­ transaminase/Alanine Aminotransaminase (AST/ALT or De-Ritis) ratio on clinically meaningful endpoints in a large

European cohort of non-metastatic renal cell carcinoma (RCC) patients. Materials and Methods: Clinico-pathological data from 698 patients with non-metastatic RCC, operated between 2005 and 2013 at a single tertiary academic center, were evaluated retrospectively. The potential prognostic value of the AST/ALTratio was analyzed using the Kaplan-Meier method, as well as uni- and multivariate Cox proportional regression models. The impact of the AST/ALT-ratio on the predictive accuracy of the Leibovich prognosis score was determined by Harrell’s concordance index (c-index). Results:  An elevated (≥ 1.26) pre-operative AST/ALT-ratio was statistically significantly associated with several wellestablished prognostic factors, including pathologic T-stage, as well as the presence of histologic tumor necrosis (p < 0.05). In multivariate analysis, an elevated pre-operative AST/ALT-ratio was identified as an independent prognostic factor for metastasis-free- (MFS) [HR = 1.61, 95% CI = 1.25–2.07, p < 0.001], as well as for overall survival (OS) [HR = 1.76, 95% CI = 1.34–2.32, p < 0.001]. Harrell’s c-index was 0.77 using the Leibovich prognosis score and 0.81 when the AST/ALT-ratio was added. Conclusions:  In the non-metastatic RCC patient cohort studied, the pre-operatively assessed AST/ALT-ratio represented an independent prognostic factor, which might further improve the predictive accuracy of well-established prognosis scores.

P35 Survival comparison analysis of two historical cohorts of metastatic renal cell carcinoma patients (cytokine therapy vs. targeted agents) – a European single-center experience over 26 years Angelika Bezan1, Georg C. Hutterer2, Silvia Golbeck2, Edvin Mrsic2, Daniel Krieger2, Johanna Jesche2, Karl Pummer2, Richard Zigeuner2, Martin Pichler1  ivision of Oncology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Department of Urology, Medical University of Graz, Graz, Austria 1

Introduction:  By the approval of new targeted agents in 2006, the standard of therapy in metastatic renal cell carcinoma (mRCC) changed, since they demonstrated significantly improved progression-free survival (PFS) rates compared with interferon in phase-3 clinical trials. Differences in overall survival (OS) could not be proven since many patients switched to another effective substance after progression of the disease. Thus, we compared two mRCC patient cohorts in order to detect OS differences between immunotherapy and targeted therapies in a real-life population outside controlled clinical trials. Methods:  Clinico-pathological data from 594 mRCC patients, operated between 1984 and 2010 at a single tertiary academic center, were evaluated retrospectively with the null hypothesis, that there is no statistically significant difference in OS of patients treated either with interferon or targeted agents. Using electronical patient records, all data regarding the beginning, the duration, the lines and different forms of therapies were assessed. Patients’ cancer-specific survival (CSS), as well as OS were assessed using the Kaplan-Meier method, com-

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Abstracts pared with the log-rank test. A first analysis revealed results for the entire study cohort. Subsequently, outcome analyses were restricted to mRCC patients with clear cell histology only. Results:  With respect to the complete follow-up period, our results in both analyses did not show a statistically significant OS difference between the two therapy modalities. By limiting the observation period to 5 years after treatment initiation, a  statistically significantly improved median five-year OS rate (26 mo.) for clear cell mRCC patients treated with targeted agents was observed, compared with 21 mo. in the interferon group (p = 0.028). Conclusions:  Our results confirm the presumption of an improved OS in mRCC attributable to treatments with targeted agents compared with previous cytokine therapies.

P36 EVA trial: a single-arm, multicenter open-label phase-II study of paclitaxel and pazopanib in patients with locally advanced and metastatic angiosarcoma Thomas Brodowicz1,4, Wolfgang Eisterer2,4, Sophie Schur1,4, Marija Balic3,4, Daniel Pink5, Peter Reichardt5, Peter Hohenberger6, Regina Knittelfelder4, Elisabeth Lindner4, Ferdinand Ploner3,4  ivision of Oncology, Department of Medicine I, Medical D University of Vienna, Vienna, Austria 2 Department of Internal Medicine V, Medical University of Innsbruck, Innsbruck, Austria 3 Division of Oncology, Department of Internal Medicine, Medical University of Graz, Graz, Austria 4 Sarcoma Platform Austria, Vienna, Austria 5 HELIOS Klinikum Berlin-Buch, Sarcoma Center Berlin-Brandenburg, Berlin, Germany 6 Interdisciplinary Tumor Center Mannheim, Sarcoma Unit, Mannheim University Medical Center, Mannheim, Germany 1

Angiosarcoma (AS) is a rare subtype of soft tissue sarcomas (STS). In metastatic disease paclitaxel has shown to be an effective systemic first-line therapy. Pazopanib, a multi-tyrosine kinase inhibitor, has recently been approved as treatment for specific subtypes of STS. Thus, the combination of paclitaxel with the anti-angiogenic drug pazopanib might be of particular value in AS, a tumor entity originating in blood vessels. This currently recruiting phase II trial, an intergroup study of the German Interdisciplinary Sarcoma Group (GISG) and the Sarcoma Platform Austria (SPA), was designed to evaluate the efficacy of pazopanib in combination with paclitaxel in patients with locally advanced or metastatic angiosarcoma. Patients suffering from histologically confirmed angiosarcoma (primary or secondary angiosarcoma after prior irradiation) with locally advanced or metastatic disease, progressive disease within 6 months before study entry, ECOG ≤ 2, at least one measurable lesion (RECIST v.1.1) and adequate organ function can be enrolled into this study. Enrolled patients receive a combination therapy of paclitaxel 70 mg/m2/d on days 1, 8 and 15, q28d x 6 and pazopanib 800mg/d po daily. In case of absence of disease progression after 6 cycles pazopanib is given as monotherapy until disease progression, unacceptable toxicity, withdrawal or death.

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The primary endpoint of this study is progression free survival. Secondary endpoints include overall survival, objective response rate and toxicity. Seven sites in Germany and three sites in Austria participate in this study. Four patients have been enrolled so far. Last Patient In is expected in Q4/2018.

P37 Chronic lymphocytic leukemia induces an exhausted T cell phenotype in the tcl1transgenic mouse model Kemal Catakovic1,2, Franz Gassner1,2, Nadja Zaborsky1,2, Stefan Rebhandl1,2, Michael Huemer1,2, Alexander Egle1,2, Tanja Hartmann1,2, Richard Greil*,1,2, Roland Geisberger*,1,2  IMCR, 3rd Medical Department, SALK, Paracelsus Private L Medical University, Salzburg, Austria SCRI, 3rd Medical Department, SALK, Paracelsus Private Medical University, Salzburg, Austria * equally contributing authors

1

2

Numerous studies show that T cells are implicated in disease progression of B cell chronic lymphocytic leukemia (CLL). Recently T cell exhaustion, which is characterized by poor effector function and increased expression of inhibitory receptors, has been observed in CLL. In this study, we investigate the role of T cell exhaustion in the tcl1tg mouse model for CLL. Our findings reveal, that T cell exhaustion in tcl1tg mice is similar to human CLL with high expression of PD-1 and Lag-3 on T cells and PD-ligand 1 (PD-L1) on CLL cells. Additionally, exhausted T cells increased with CLL progression. Furthermore, inhibition of PD-1/PD-L1 reactivated exhausted T cells towards cytotoxicity. Our data revealed, that T cell exhaustion contributes to CLL pathogenesis and that blockade of the PD-1/PD-L1 interaction represents a novel therapeutic approach for CLL.

P38 DIEPP: Prospektive Evaluierung zu Dosisintensität und Neutropenieprophylaxe bei Krebspatienten mit einem hohen Risiko einer Chemotherapie-induzierten febrilen Neutropenie (FN) Ferdinand Haslbauer1, Wolfgang Willenbacher2, Rainer Kolb3, Arik Galid4, Ernst Ulsperger5, Christine Staudigl6, Peter Neumeister7, Christine Jäger8, Christian Singer6 Interne Medizin, Landeskrankenhaus Vöcklabruck, Vöcklabruck, Österreich 2 Innere Medizin V (Hämatologie & Onkologie), Medizinische Universität Innsbruck, Innsbruck, Österreich 3 Pulmologische Abteilung, Klinikum Wels-Grieskirchen, Wels, Österreich 4 Ordination, Wien, Österreich 5 Ordination, Eggenburg, Österreich 1

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Abstracts  niversitätsklinik für Frauenheilkunde, Medizinische U Universität Wien, Wien, Österreich 7 Klinische Abteilung für Hämatologie, Medizische Universität Graz, Graz, Österreich 8 Amgen GmbH, Wien, Österreich 6

Einleitung und Fragestellung: Bestimmte Chemotherapien, sowie gewisse Patientencharakteristika gehen mit einem hohen FN-Risiko einher. Pegfilgrastim ist zur FN-Prophylaxe bei Krebspatienten unter myelosuppressiver Chemotherapie zugelassen. Studienziele:  Erhebung der FN-Inzidenz unter Pegfilgrastim Primärprophylaxe (PP) oder Sekundärprophylaxe (SP), der relativen Chemotherapie-Dosisintensität (RDI) und der Verwendungspraxis von Pegfilgrastim bei Patienten mit hohem FN-Risiko in Österreich. Material und Methoden:  DIEPP Österreich war Teil einer internationalen, multizentrischen, prospektiven Beobachtungsstudie an erwachsenen Patienten mit Mammakarzinom, Lungenkarzinom, Kolonkarzinom, Ovarialkarzinom oder Lymphom, welche ein FN-Gesamtrisiko (chemotherapie- plus patientenbezogenes FN-Risiko) ≥ 20% hatten und eine Chemotherapie mit prophylaktischem Pegfilgrastim erhielten. Ergebnisse:  Zwischen August 2010 und Juli 2013 wurden 217 Patienten in 17 Zentren eingeschlossen und ausgewertet. Die häufigsten Tumortypen waren Mammakarzinom (59%, n = 128), Lymphom (20%, n = 43) und Lungenkarzinom (15%, n = 33). Patienten mit chemotherapiebezogenem FN-Risiko ≥ 20% erhielten zu 91% (n = 135/148) eine PP, bei 10-19% zu 71% (n = 63/89). Die wichtigsten Indikationen für die Pegfilgrastim-Gabe waren geplante Chemotherapie mit hohem FN-Risiko, weibliches Geschlecht, fortgeschrittene Erkrankung und Alter ≥ 65 Jahre. Die FN-Inzidenz war insgesamt 9% (n = 20), 7,5% bei PP-Patienten (n = 12/159) und 14% (n = 7/49) bei SP-Patienten; bei 6 der 7 SP-Patienten trat die FN vor erstmaliger Pegfilgrastim-Gabe auf. FN verursachte 43% (n = 22/51) der Hospitalisierungen. Insgesamt erhielten 78% (n = 100/128) der Mammakarzinompatientinnen eine RDI ≥ 85% und 54% (n = 23/43) der Lymphompatienten eine RD ≥ 90%. 8 Patienten (4%) hatten insgesamt 16 unerwünschte Arzneimittelwirkungen, keine davon schwer. Am häufigsten waren Knochenschmerzen (5 Ereignisse) und Ödeme (3 Ereignisse). Schlussfolgerungen:  FN war in dieser FN-Hochrisikopopulation unter Pegfilgrastim Prophylaxe relativ selten, verursachte jedoch > 40 % aller Hospitalisierungen. FN-Prophylaxe erfolgte gemäß Guidelines. Sponsor: Amgen.

P39 GIRK1: a new prognostic biomarker for estrogen receptor positive breast cancer Sarah Kammerer1, Armin Sokolowski1, Hubert Hackl2, Dieter Platzer1, Stephan Jahn3, Florentia Peintinger3, Wolfgang Schreibmayer1, Thomas Bauernhofer4  epartment of Biophysics, Medical University of Graz, Graz, D Austria 2 Division of Bioinformatics, Biocenter, Medical University of Innsbruck, Innsbruck, Austria 3 Department of Pathology, Medical University of Graz, Graz, Austria 1

Division of Oncology, Department of Internal Medicine, Medical University of Graz, Graz, Austria

4

Introduction:  Overexpression of GIRK1 (G protein-activated inward rectifier potassium channel 1) in tumours of breast cancer patients has been correlated with lymph node metastases and poor prognosis in previous studies with small patient numbers. The aim of this study was to validate the use of GIRK1 as prognostic breast cancer biomarker. Methods:  Gene expression data of breast carcinoma available from the cancer genome atlas (TCGA, n = 905) and a data set of estrogen receptor (ER) positive breast cancer patients (GEO-ID: GSE17705, n = 298) were analysed using SigmaStat/ SigmaPlot, R and Genesis. Results:  We found that GIRK1 mRNA expression levels were significantly higher in breast tumours compared to corresponding normal tissue (TCGA, n = 105, p < 0.001). GIRK1 was significantly overexpressed in tumours of ER positive patients compared to ER negative patients (TCGA, n = 841, p < 0.001). Spearman rank correlation and hierarchical cluster analysis strongly supported the link between GIRK1 overexpression and ER positive breast cancer subtypes in both data sets studied. Amongst the ER positive patients, those with high GIRK1 levels had a worse overall survival probability than patients with lower GIRK1 levels (TCGA, n = 609, p < 0.05, HR = 1.71). Interim analysis of the second data set showed a trend towards worse overall survival probability for patients with high GIRK1 levels (n = 99, p = 0.064, HR = 1.7). Conclusions:  The analysis of two large gene expression sets indicates that GIRK1 mRNA overexpression might serve as new prognostic biomarker for ER positive breast cancer patients. We are planning to establish an RNA in situ hybridization protocol for GIRK1 to be used in daily routine.

P40 Gene signature indicates different antineoplastic activities of statins and bisphosphonates Heidrun Karlic1, Florian Haider2, Silvia Spitzer2, Franz Varga2 1 2

Ludwig Boltzmann Cluster Oncology, Vienna, Austria Ludwig Boltzmann Institute of Osteology at the Hanusch Hospital of WGKK and AUVA, Vienna, Austria

The aim of this study was to identify the molecular mechanisms and biological pathways associated with the anticancer effects of statins and bisphosphonates, which are known to downregulate the farnesylation and geranyl-geranylation of essential membrane-associated signal-transducers such as RAS and RHO proteins. Transcriptomic, proteomic and methylomic analyses were done from the neoplastic cell lines MDAMB-231 breast cancer, PC-3 prostate carcinoma, MG-63 and U2-OS osteosarcoma and HMC-1 mast cell leukemia being treated for 3 days with pharmacologic doses with a representative statin (simvastatin) and a bisphosphonate (ibandronate). Bioinformatic analyses involved the gene set enrichment analysis (GSEA) and Pathvisio software as pathway recognition algorithms. Statin-treatment stimulated markers from mesenchymal stem cells such as SOX2 and various collagens, thus indicating a tendency towards EMT (epithelial mesenchymal transition). The gene sets predominantly targeted by

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Abstracts the bisphosphonate were associated with a re-activation of the TP53 tumor suppressor gene and associated pathways of DNAdamage and –repair. Both drugs also downregulated the EZH2 histone methylase and associated genes from the PRC (polycomb related complex), thus promoting EMT. In addition, the analysed profiles displayed some similarities to that of histone deacetylase (HDAC) inhibitor-treated cells. Differences in drug sensitivity between cell types were associated with dysregulation of marker proteins from potassium voltage-gated channels from respective cellular membranes, whose components may rely on geranyl-geranylation and farnesylation, the latter being targeted by both statins and bisphosphonates. This provides some explanation for clinical observations, by indicating shared mechanisms for the anti-cancer and anti-osteoclastic activities of statins and bisphosphonates.

P41 Development of a 3D cell culture model for the investigation of cancer cell/immune cell interactions Stefan Köck1, Marit Zwierzina2, Julia Magdalena Huber1, Mario Britsche2, Gabriele Gamerith1, Edith Lorenz1, Annabella Knab2, Heinz Zwierzina1, Arno Amman1  epartment of Internal Medicine V, Medical University of D Innsbruck, Innsbruck, Austria 2 Department of Anatomy and Histology, Medical University of Innsbruck, Innsbruck, Austria 1

Introduction:  Recent studies have shown the obvious interaction between cancer cells and immune cells. These effects can be hardly studied in conventional culture systems using cell monolayers. Therefore, alternative in vitro models are required. We describe a novel 3D cell culture model and a 3D migration assay for investigating cancer cell/immune cell interactions. Methods:  A multi-well hanging drops system was used to produce 3D tumor spheres. The human NSCLC cell lines A549, Calu-6 and Colo699 were incubated for 6 and 7 days in the hanging drops to form spheroids. On day 5, peripheral blood mononuclear cells (PBMC) were added either with or without interleukin-2 (IL-2). Viability was investigated via flowing cytometry. Invasion of PBMC into the tumor spheres was measured by performing immunohistochemistry. Results:  No effect on cell viability was observed in A549 and Colo699 spheroids. In Calu-6 spheroids, a significant PBMC induced cytotoxic effect was measured, what was even stronger under IL-2 stimulation. Immunohistochemical staining revealed PBMC infiltration in all three cell lines. Under IL-2 stimulation, infiltration of PBMC increased in Calu-6 and Colo699 spheroids. Conclusions:  This work provides evidence that our 3D coculture model is a reliable and effective approach to study interactions between cancer cells and immune cells. Additionally, we were able to establish for the first time an innovative ex vivo infiltration assay based on 3D microtissues.

P42 Depression in elderly cancer patients – associations with geriatric syndromes and chemotherapy Hendrik Koller1, Elisa Lengauer2, Georg Schreil2, Ansgar Weltermann2 1

Introduction:  Depression is a comorbid disabling syndrome that commonly affects cancer patients. The purpose of our study was to examine prevalence of depression in elderly cancer patients and possible associations between demographic and geriatric variables. Furthermore, we were interested whether the presence of depression was associated with lower rates of chemotherapy, chemotherapy completion rates, delivered dose density and re-hospitalization. Methods:  From March 2010 to October 2013, patients > 70 years with a diagnosis of a hematologic or solid malignancy were screened for the presence of depression, defined by a score of 5 or more points on the geriatric depression scale (15 item version). Prevalence rate and possible associations between depression and various components of the geriatric assessment and demographic date were examined. Furthermore, a subgroup of colorectal cancer patients was analyzed for a possible relationship between the chemotherapy and depression. Results:  Overall, geriatric depression scores were available for 174 patients. Depression, defined by a score of 5 points or more was present in 35 patients (20%); 48 patients (28%) already received treatment with antidepressants. GDS scores above 5 were associated with female sex, ADL and IADL impairment, nutritional status, cognitive function, falls, physical performance measures, number of geriatric syndromes present, frailty and ECOG status (p < 0.001). In a subgroup of 47 CRC patients, we found no difference in chemotherapy completion rates, delivered dose density and re-hospitalization rate between depressed and non-depressed patients. Conclusions:  Depression is common in elderly cancer patients and associated with a large number of demographic and geriatric variables, implying significant impact on quality of life of these patients. However, completion of chemotherapy, delivered dose density and re-hospitalization rate were independent of the presence of depressive symptoms.

P43 Laboratory parameters have prognostic value in patients with newly diagnosed brain metastases: analysis of 1201 cases Romina Koller1,2, Anna Sophie Berghoff1,2, Georg Widhalm2,3, Karin Dieckmann2,4, Christoph Zielinski1,2, Peter Birner2,5, Rupert Bartsch1,2, Matthias Preusser1,2 1

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 epartment Akutgeriatrie, Department of Internal Medicine D III, Hospital Elisabethinen Linz, Linz, Austria Department of Internal Medicine I, Hospital Elisabethinen Linz, Linz, Austria

2

 epartment of Medicine I, Medical University of Vienna, D Vienna, Austria

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Abstracts Comprehensive Cancer Center, Medical University of Vienna, Vienna, Austria 3 Department of Neurosurgery, Medical University of Vienna, Vienna, Austria 4 Department of Radiotherapy, Medical University of Vienna, Vienna, Austria 5 Department of Pathology, Medical University of Vienna, Vienna, Austria 2

Introduction:  We investigated the prognostic value of laboratory parameters in a large real life cohort of brain metastasis (BM) patients. Methods:  1201 Patients treated for newly diagnosed BM from extracranial solid cancers (472/1201 (39.3%) lung cancer; 266/1201 (22.1%) breast cancer; 163/1201 (13.6%) melanoma; 121/1201 (10.1%) renal cell carcinoma; 101/1207 (8.4%) colorectal cancer; 78/1201 (6.5%) other primary tumor) were included in the analysis. Clinical characteristics including laboratory parameters at diagnosis of BM and survival times were retrieved by chart review. Laboratory parameters with significant impact on overall survival from diagnosis of BM in univariate analysis were entered in multivariable analysis together with known prognostic factors including primary tumor type, graded prognostic assessment (GPA) class and treatment modality for BM. Results:  At univariate survival analysis, hemoglobin level <  lower limit of normal (LLN), platelet count <  LLN, leukocyte count < LLN, serum albumin level < LLN, serum c-reactive protein (CRP) concentration > upper limit of normal (ULN), serum creatinine concentration > ULN and serum lactate dehydrogenase level (LDH) > ULN correlated with unfavourable overall survival (p < 0.05, log-rank test). At multivariate analysis, hemoglobin (HR 0.7; p = 0.001), platelet count <  LLN (HR 0.8; p = 0.024), leukocyte count <  LLN (HR 1.2; p = 0.039), albumin (HR 0.8; p = 0.032), CRP (HR 1.6; p <  0.001) and LDH (HR 1.3; p = 0.001) concentrations retained their statistically significant impact on overall survival. Conclusions:  Blood hemoglobin level, serum albumin concentration, serum CRP concentration, and serum LDH concentration have a strong and independent prognostic impact in patients with newly diagnosed BM and should be incorporated in prognostic scores.

P44 CYP39A1 polymorphism protects against toxicity during intensive induction chemotherapyin patients with advanced head and neck cancer Thomas Melchardt1, Clemens Hufnagl1, Teresa Magnes1, Gerhard Moser2, Alexander Schlattau3, Daniel Neureiter4, Wolfgang Tränkenschuh4, Lukas Weiss1, Richard Greil1, Alexander Egle1  epartment of Internal Medicine III, Paracelsus Private D Medical University, Salzburg, Österreich 2 Department of Otorhinolaryngology, Paracelsus Private Medical University, Salzburg, Österreich 3 Institute of Radiology, Paracelsus Private Medical University, Salzburg, Österreich 1

Institute of Pathology, Paracelsus Private Medical University, Salzburg, Österreich

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Introduction:  The toxicity of intensive induction chemotherapy including cisplatin, docetaxel and 5-fluoruracil is a major obstacle to its use in patients with advanced head and neck cancer. In particular, docetaxel significantly increases the rate of leucopenia. Genetic polymorphisms involved in drug metabolism may help to identify patients at high risk for treatment related toxicity. Genetic polymorphism of the CYP39A1 gene has been described to be associated with docetaxelinduced leucopenia in gynecological malignancies. Methods:  A cohort of 78 patients with advanced head and neck cancer consecutively treated with intensive induction chemotherapy were assessed for clinical outcome and toxicity. Genetic polymorphisms of 5 genes known to be associated with efficacy and 1 gene associated with toxicity of this treatment were analyzed in all patients. Results:  Median progression free survival and overall survival was 20 and 31 months in an intention to treat analysis, respectively. Overall response rate was high with 78.1% and 88.5% of the patients were able to receive subsequent radiotherapy as planned. No tested genotype affected efficacy, but genotype A of the CYP39A1 rs7761731 polymorphism was associated with a higher incidence of toxicity during treatment. Conclusions:  Intensive induction chemotherapy results in a high response rate in the majority of patients with advanced head and neck cancer and. The CYP39A1 polymorphism rs7761731 may help to identify patients at high risk for treatment related toxicity.

P45 Mitogen-inducible gene-6 inhibits malignant transformation of mutated BRAF by mediating feedback inhibition towards the epidermal growth factor receptor Malgorzata Milewska1, David Romano1, Maria Luisa Guerriero1, Marc Birtwistle2, Franz Quehenberger3, Stefan Hatzl4, Ana Herrero-Mier1, Boris N. Kholodenko1, 5, 6, Oreste Segatto7, Walter Kolch1,5,6, Armin Zebisch4  ystems Biology Ireland, University College Dublin, Dublin, S Ireland 2 Mount Sinai Hospital, New York, USA 3 Institute of Medical Informatics, Medical University of Graz, Graz, Austria 4 Division of Hematology, Department of Internal Medicine, Medical University of Graz, Graz, Austria 5 Conway Institute of Biomolecular & Biomedical Research, University College Dublin, Dublin, Ireland 6 School of Medicine and Medical Science, University College Dublin, Dublin, Ireland 7 Laboratory of Immunology, Regina Elena Cancer Institute, Rome, Italy 1

Introduction:  BRAF mutations frequently occur in human cancers and transform cells by activating the RAS-extracellular signal-regulated kinase (ERK) signaling cascade. Interestingly, however, activation of this pathway may also cause a negative feedback that inhibits the epidermal growth factor receptor (EGFR) and thereby limits its own oncogenic potential. We

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Abstracts studied the role of mitogen-inducible gene-6 (MIG-6), a potent EGFR inhibitor, within this process. Materials and Methods:  qPCR and Western blot analyses were used to study the expression and phosphorylation of intracellular signaling modules in A431, Cos-1 and MCF-7 cell lines following transfection with various BRAF mutations + /− MIG-6 as well as after incubation with pharmacological inhibitors for MEK and EGFR. The same settings were used for NIH3T3-focus formation assays. The clinical relevance of these findings was evaluated in a database retrieval of 394 patients with papillary thyroid cancer (PTC), an entity characterized by the frequent occurrence of mutations in BRAF. Results:  Mutated BRAF causes a RAS-ERK mediated induction of MIG-6 expression, which in turn inhibits activation of the pEGFR. Consistently, modulation of MIG-6 expression regulates the oncogenic potential of mutant BRAF via EGFR downregulation in focus formation assays, indicating that MIG-6 plays a central role within feedback inhibition from BRAF towards the EGFR. These data are also relevant for PTC patients, as we observed a more aggressive disease phenotype in cases where mutant BRAF coexisted with low MIG-6 expression. Conclusions:  These data demonstrate that MIG-6 efficiently reduces the oncogenic potential of mutated BRAF by orchestrating a negative feedback circuit directed towards the EGFR.

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P47 Spinophilin expression determines cellular growth, cancer stemness and 5-flourouracil resistance in colorectal cancer Anna Lena Ress1, Verena Stiegelbauer1, Daniela Schwarzenbacher1, Alexander Deutsch2, Hui Ling3, George Adrian Calin3, Beate Rinner4, Armin Gerger1, Hellmut Samonigg1, Martin Pichler1,3  ivision of Oncology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Division of Heamatology, Department of Internal Medicine, Medical University of Graz, Graz, Austria 3 Department of Experimental Therapeutics, The University of Texas, Houston, USA 4 Center for Medical Research, Medical University of Graz, Graz, Austria 1

Stage specific miRNA expression in colon cancer Melanie Rammer1, Gerald Webersinke1, Sophie Haitschi-Petnehazy2, Eva Bauer2, Hubert Hackl3, Pornpimol Charoentong3, Zlatko Trajanoski3, Theodora Malli1, Andreas Petzer4, Holger Rumpold4  aboratory for Molecular Biology, Internal Medicine I, L Hospital Barmherzige Schwestern, Linz, Austria 2 Department of Pathology, Hospital Barmherzige Schwestern, Linz, Austria 3 Biocenter Innsbruck, Section for Bioinformatics, Medical University of Innsbruck, Innsbruck, Austria 4 Department of Internal Medicine I, Hospital Barmherzige Schwestern, Linz, Austria 1

Introduction:  In recent years studies have confirmed the regulatory potential of microRNAs. Alterations of miRNA expression can be associated with cancer types and could have a prognostic and predictive value. Aim of the study is to evaluate miRNA expression patterns correlating with colon carcinoma stages. Along with comparing primary tumor tissue from lymph node negative with lymph node positive tumors, the question of T stage dependent miRNA expression is addressed. Material and Methods:  Total RNA was extracted from 170 FFPE samples using the RecoverAll FFPE kit (Ambion). miRNA array analysis (Affymetrix) was conducted for 40 test samples, investigating expressions of 1,733 human mature miRNAs. For array data validation of selected miRNAs in a larger cohort, qPCR was carried out using TaqMan miRNA assays (LifeTechnologies). Results:  Array data of test samples and qPCR data of validation samples comparing lymph node positivity and negativity showed no significantly differentially expressed miRNAs. Anal-

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ysis of array data considering differences between the T stages (acc. to UICC criteria) displayed significant differences. qPCR validation analysis revealed interesting results for several miRNAs, e.g. miR-18a or miR-378a. Conclusions:  Regarding miRNA expression according to lymph node status, array analyses as well as validating qPCR analyses do not display significant differences. According to T stage however, both methods reveal significantly differing miRNA levels indicating miRNA involvement in tumor progression. This should be further validated in larger cohorts and could broaden our understanding of the role of miRNAs in disease biology to facilitate accurate stage determination in the future.

Introduction:  The putative tumor suppressor gene spinophilin has been involved in cancer progression in several types of cancer. In this study, we explored the role and prognostic value of spinophilin in colorectal cancer. Material and Methods:  We explored the prognostic value of spinophilin expression in 162 colon adenocarcinoma patients. In addition, we generated stably expressing spinophilindirected shRNA CRC cell lines and studied the influence of spinophilin expression on cellular phenotypes and molecular interactions. Results:  We independently confirmed that low spinophilin expression levels are associated with poor prognosis in CRC patients (p = 0.038). A reduction of spinophilin levels in p53 wild-type HCT116 and p53-mutated Caco-2 cells led to increased cellular growth rates and anchorage-independent growth (p < 0.05). At molecular level, reduced spinophilin levels increased the expression of the transcription factor E2F-1. In addition, we observed an increased formation of tumor spheres, increased number of CD133 positive cells and an increased resistance to 5-flourouracil (p < 0.05). Finally, treatment with the de-methylating agent 5-aza-dC increased spinophilin expression in CRC cells (p < 0.05), corroborated by a correlation of spinophilin expression and extent of methylated CpG sites in the gene promoter region (p < 0.001). Conclusions:  Gain of aggressive biological properties of CRC cells including cellular growth, cancer stem cell features and 5-flourouracil resistance partly explains the role of spinophilin in CRC.

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Abstracts

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INSIGHT Österreich: Baseline-Daten zur Therapie des hepatozellulären Karzinoms (HCC) mit Sorafenib Walter Spindelböck , Robert Buder , Ivo Graziadei , Markus Peck-Radosavljevic4, Felix Keil5, Christoph Tinchon5, Franziska Van Zijl6, Rudolf E. Stauber1 1

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 linische Abteilung Gastroenterologie und Hepatologie, K Universitätsklinik für Innere Medizin, Medizinische Universität Graz, Graz, Österreich 2 Gastroenterologie, Innere Medizin, Krankenhaus der BHB Linz, Linz, Österreich 3 Klinische Abteilung für Gastroenterologie, Universitätsklinik für Innere Medizin, Medizinische Universität Innsbruck, Innsbruck, Österreich 4 Klinische Abteilung für Gastroenterologie und Hepatologie, Universitätsklinik für Innere Medizin, Medizinische Universität Wien, Wien, Österreich 5 Department Hämato-Onkologie, Landeskrankenhaus Leoben, Leoben, Österreich 6 Bayer Austria GmbH, Wien, Österreich 1

Einleitung:  Sorafenib ist beim fortgeschrittenen HCC die bisher einzige Substanz, für die eine signifikante Lebensverlängerung in RCTs gezeigt werden konnte. Im Rahmen der Phase IV – Studie INSIGHT wurden 782 Patienten prospektiv an 124 Zentren in Österreich und Deutschland dokumentiert. Methoden:  Es konnten HCC Patienten, welche für eine Therapie mit Sorafenib geeignet waren, eingeschlossen werden. Hier wird erstmals die österreichische Subpopulation der INSIGHT-Studie zum Studieneinschluss (Baseline) dargestellt. Ergebnisse:  Es wurden im Zeitraum von 2007-2013 an 21 Zentren 139 auswertbare Patienten (18% der Gesamtstudie) eingeschlossen. Das Alter lag bei 64 ± 9 Jahren, 87% der Patienten waren männlich. Die mittlere Startdosis war bei Patienten unter 65 Jahren mit 736 ± 144 mg höher als bei älteren Patienten (660 ± 191 mg). 53%, 37% und 9% der österreichischen Patienten wiesen einen ECOG Status von 0, 1 und ≥ 2 zu Studieneinschluss auf. Dabei waren 55%, 32% und 9% in den Child-Pugh Stadien A,B und C. Gemäß den EASL-EORTC Richtlinien entsprachen 9%, 25%, 60% und 4% den BCLC Stadien A, B, C und D. Als Vortherapien wurden Operation bei 14%, Radiofrequenzablation oder Ethanolinjektion (RFA/PEI)bei 17% und TACE bei 35% der Patienten durchgeführt. Schlussfolgerung:  Prospektive real-life Daten zur Therapie des fortgeschrittenen HCC mit Sorafenib in Österreich existierten bisher nicht. Die in Österreich gesammelten Daten von 139 Patienten wiesen zu Studieneinschluss eine für die Indikation charakteristische Alters- und Geschlechtsverteilung auf. 60% der Patienten befanden sich in der empfohlenen BCLC Kategorie C. Bemerkenswert sind der im Vergleich zur Gesamtpopulation bessere Leistungsstatus, die fortgeschrittenere Lebererkrankung und das unterschiedliche Profil an Vortherapien der Patienten.

Cancer stem cell gene variants and colon cancer prognosis Michael Stotz1, Sereina A. Herzog2, Martin Pichler1, Joanna Szkandera1, Renate Schaberl-Moser1, Hellmut Samonigg1, Wilfried Renner3, Andrea Berghold2, Armin Gerger1  ivision of Oncology, Department of Internal Medicine, D Medical University of Graz, Graz, Austria 2 Institute for Medical Informatics, Statistics and Documentation, Medical University of Graz, Graz, Austria 3 Clinical Institute of Medical and Chemical Diagnostics, Medical University of Graz, Graz, Austria 1

Introduction:  Growing evidence suggests that human cancers are stem cell diseases. Recently, the hypothesis that a specific subset of cancer stem cells (CSC) is responsible for metastasis has been investigated in CSCs isolated from human colon cancer specimens. There is substantial germline genetic variability within the stem cell genes DPP4 (CD26), PROM1 (CD133) and CD44, including multiple single nucleotide polymorphisms (SNPs). In this study we investigated 9 CSC gene variants (rs187116, rs7116432, rs2268889, rs353639, rs3788979, rs13347, rs187115, rs7608798, rs2240688) in DPP4, PROM1 and CD44 to predict the clinical outcome in stage II and III colon cancer patients. Methods:  599 patients with stage II and III colon cancer were included in this retrospective study. The associations of time to recurrence (TTR) and overall survival (OS) with patient's clinicopathological characteristics were assessed using univariate Cox-regression analysis. The association between each polymorphism and TTR/ OS was examined using Kaplan–Meier curves and log-rank test. Hazard ratio (HR) and 95% confidence interval (CI) are displayed using univariate Cox-regression analysis. Results:  The associations of TTR/OS with patient's clinicopathological characteristics are shown in Table 1. Genotype frequencies were: CC 58.5%, CT 35.9% and TT 5.6% for rs13347 and GG 13.4%, GA 48.9% and AA 37.7% for rs187115 in CD44. A significant association was found between CD44 rs13347 (HR = 0.62 (95 % CI 0.42-0.93), p = 0.019) and OS and CD44 rs187115 (HR = 0.46 (95 % CI 0.24-0.87), p = 0.014) with TTR. No significant association was found between the other SNPs and TTR/ OS. Multivariate analyses are ongoing. Conclusions:  Our data suggest that the germline variants rs13347 and rs187115 in the stem cell gene CD44 are prognostically relevant in stage II and III colon cancer patients.

P50 Quantitative Erfassung des psychoonkologischen und psychosozialen Unterstützungsbedarfs onkologischer Patienten mittels validiertem Hornheider-Fragebogen Petra Sumnitsch1,3, Bernd Hartmann1, Daniela Zanolin2,3, Christoph, H. Saely1,2,3, Alois Lang1 1

 bteilung für Innere Medizin und Kardiologie am A Akademischen Lehrkrankenhaus, Feldkirch, Austria

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Abstracts Vorarlberg Institute for Vascular Investigation and Treatment (VIVIT), Feldkirch, Austria 3 Private Universität Fürstentum Liechtenstein, Triesen, Liechtenstein 2

Einleitung und Fragestellung: Die Diagnose einer onkologischen Erkrankung bedeutet für Betroffene eine starke psychische Belastung. Ein Bedarf an professioneller psychoonkologischer bzw. psychosozialer Betreuung erscheint naheliegend. Ziel dieser Studie ist die quantitative Erfassung dieses Bedarfs. Material und Methoden: Der psychoonkologische bzw. psychosoziale Betreuungsbedarf wurde bei 200 konsekutiven Patienten mittels des validierten Hornheider Fragebogens zweimalig im Abstand von 6 Monaten untersucht. Ein Summenscore von ≥ 4 im Hornheider Fragebogens zeigt Unterstützungsbedarf an. Zwischen den Erhebungen wurden psychoonkologische Beratungsgespräche angeboten. Ergebnisse:  Von insgesamt 200 ausgehändigten Frage­ bogen, wurden bei der ersten Befragung 149 (68 von Frauen und 81 von Männern) retourniert, entsprechend einer Rücklaufquote 74,4%. Insgesamt hatten 33,1% der Befragten einen psychoonkologischen Unterstützungsbedarf, ohne signifikanten Unterscheid zwischen Frauen und Männern (33,3% vs. 29,4%; p = 0,608). Bei der zweiten Befragung waren von den in die Studie Aufgenommenen noch 52 Patientinnen und 49 Patienten am Leben. Der Unterstützungsbedarf ist um 3,3% (40,4% bei Frauen und 32,7% bei Männern; p = 0,420) gestiegen. Schlussfolgerungen:  Die Ergebnisse belegen den großen psychoonkologischen und psychosozialen Unterstützungsbedarf onkologischer Patientinnen und Patienten. Ein systematisches Screening könnte eine entscheidende Verbesserung der psychoonkologischen und psychosozialen Betreuungsbedürftigkeit bringen.

P51 Rasche Remission mit Immutherapie alleine bei fortgeschrittenen onkologischen Erkrankungen (2 Fallbeispiele) Daniela Voskova, Daniel Lenger, Michael A. Fridrik Interne III-Onkologie, Allgemeines Krankenhaus Linz, Linz, Österreich Einleitung:  Die monoklonalen Antikörper bedeuten einen Durchbruch in Behandlung der malignen Erkrankungen. Um eine Remission bei metastasierten soliden Tumoren oder systemischen hämatologischen Erkrankungen zu erreichen, ist meistens eine zusätzliche Chemotherapie erforderlich. An folgenden 2 Beispielen präsentieren wir potente Wirkung der Antikörpertherapie alleine bei fortgeschrittenen malignen Erkrankungen. Fallbeispiel 1: 47-jährige Patientin: 01/2014 Diagnose eines Mammakarzinoms mit Lebermetastasen, Lymphknotenmetastasen, Knochenmetastasen und Knochenmarkskarzinose. Histologie: apokrines Karzinom, Hormonrezeptoren negativ, Her-2 3+ , Ki 67 30%. Rasch progredientes Ikterus; ECOG 2. Labor: Hb 8,9 g/dl; AST 4149 U/l; ALT 766 U/l; g-GT 1047 U/l; ALP 811 U/l; Bilirubin 8,3 mg/dl. Leber MR: multiple Lebermetastasen. 28.1.2014 -1.4.2014 Immuntherapie Trastuzumab und Pertuzumab. Labor: Hb 9,8 g/dl; AST 112 U/l;; ALT 65 U/l; g-GT 266 U/l;; ALP 271 U/l;; Bilirubin 0,8 mg/dl. Leber

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MR nach 3 Kursen: keine Lebermetastasen. 1.4.-15.7.2014 zusätzlich Docetaxel (6 Kurse). Seit 08/2014 komplette Remission; ECOG 0; Labor: Hb und LFP im Normbereich. Fallbeispiel 2: 28-jähriger Patient:  11/2009 Diagnose eines großzelligen anaplastischen T-Zell Lymphoms CD 30- und ALK -positiv; Stadium IIA. 11/2009-04/2010 4 Kurse Chemotherapie Schema: CEOP/IMVP-Dexamethason. 03/2012 1.Rezidiv; Stadium IA. 03/2012-04/2012 3 Kurse Salvagetherapie Schema: ESHAP. 05/2012 Hochdosistherapie nach dem BEAM Protokoll gefolgt von autologer Stammzelltransplantation. 11/2014 2. Rezidiv; Stadium IV: Knochenmarksbefall. Lymphknotenbefall infrakarinal, hilär, mesenterial und retroperitoneal. 11/201412/2014 3 Kurse Immuntherapie mit Brentuximab-Vedotin. PET-CT 01/2015 nahezu komplette Remission (Restspeicherung Humerus und Os sacrum). Schlussfolgerungen:  Die monoklonalen Antikörper können in Einzelfällen als Monotherapie eine rasche Remission erzielen und dadurch weitere wirksame Therapie bis zur Heilung ermöglichen.

P52 The novel fatty acid synthase inhibitor G28UCM promotes cancer cell death by targeting mitochondrial and metabolic pathways Nastasia Wilfinger, Magdalena Billerhart, Thomas Grunt, Karin Nowikovsky Department of Medicine I, Medical University of Vienna, Vienna, Austria In general, healthy cells rely on exogenous lipids rather than on fatty acid synthesis, whereas tumour cells synthesize abundant amounts of fatty acids, using lipogenic enzymes including the fatty acid synthase (FASN). As the overexpression of FASN represents a characteristic of a variety of tumour cells, targeting FASN became a novel appealing strategy of cancer treatment. Most recently, a natural derivative of epigallocatechin gallate, G28UCM, has been developed by Teresa Puig et.al. (2009). In this study we investigated the relevant target genes involved in the mode of action of G28UCM, focussing on mitochondrial and metabolic signalling.We used SKOV-3 cells as ovarian cancer model system, and applied following methods: real-time monitoring of cell viability (xCElligence), detection of respiration and extracellular flux (Seahorse analyser), ROS (flow cytometry), protein and gene expression (western blotting and RT-PCR) and immunofluorescence (confocal scanning microscopy).Our data indicates that G28UCM rapidly modified the cellular metabolism and induced cell death within 48 to 72 hours. SKOV-3 cells became highly glycolytic under drug exposure and displayed reduced mitochondrial oxidative phosphorylation. Interestingly, withdrawal of glucose exhausted the glycolytic reserve, indicating that no other nutrient source was used for energy conversion. Thus, glucose deprivation becomes detrimental to SKOV-3 cells. We also showed that blocking the FASN was accompanied by ROS formation and upregulation of HO-1. Importantly, we detected an early drastic upregulation of the stress-response protein REDD-1.Based on our findings, future studies will investigate the potential role of REDD-1 in controlling the metabolic shift induced by FASN inhibition.

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