„Rak nerki, rak prostaty 2015 – Nowe wyzwania”

KSIĄŻKA ABSTRAKTÓW

SEROCK, 23−24 października 2015 r.

Niniejsze materiały konferencyjne zostały przygotowane na podstawie abstraktów nadesłanych przez uczestników Międzynarodowego Sympozjum „Rak nerki, rak prostaty 2015 – Nowe wyzwania”

Informacje Ogólne o Sympozjum Organizator: Fundacja Onkologii Doświadczalnej i Klinicznej Termin: 23−24 pażdziernika 2015 r. Miejsce: Hotel Narvil, ul. Czesława Miłosza 14a 05-140 Serock Strona internetowa Sympozjum: www.sympozjum-raknerkirakprostaty.pl Biuro Sympozjum: Elżbieta Bernatowicz tel. kom. +48 601 367 981 Anna Sikora tel. kom. +48 530 327 947 ul. Mokotowska 4/6 00-641 Warszawa e-mail: [email protected] Wydawnictwo: Fundacja Onkologii Doświadczalnej i Klinicznej Projekt graficzny okładki: Total Project Druk: SCRIPT S.C ul. Teligi 6/22 02-777 Warszawa ISBN

978-83-938491-2-3 Sympozjum przeznaczone jest dla osób upoważnionych do wystawiania recept i osób zaopatrujących w produkty lecznicze prowadzących obrót produktami leczniczymi w rozumieniu art. 52 ust. 2 pkt 6

Ustawy z dnia 6 września 2001 r. Prawo farmaceutyczne (Dz.U. 2008, nr 45, poz. 271 ze zm.). Obowiązujące kolory smyczy: Uczestnik

Wykładowca

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Organizator

Firma

Międzynarodowe Sympozjum

„Rak nerki, rak prostaty 2015 – Nowe wyzwania” Komitet Naukowy Sympozjum: prof. dr hab. n. med. Cezary Szczylik - Przewodniczący prof. dr hab. n. med. Małgorzata Kołodziejczak prof. dr hab. n. med. Piotr Radziszewski dr n. med. Iwona Skoneczna dr hab. n. med. Rafał Stec dr hab. n. med. Gabriel Wcisło dr n. med. Anna Czarnecka dr n. med. Anna Kulik dr n. med. Wojciech Rogowski dr n. med. Piotr Tomczak Komitet Organizacyjny Sympozjum: dr inż. Elżbieta Bernatowicz, e-mail: [email protected]; tel. kom. 601-367-981 mgr Agata Kiełkiewicz, e-mail: [email protected], tel. kom. 697-010-206 mgr Aleksandra Klemba, e-mail: [email protected]; tel. kom. 502-670-949 mjr lek. Przemysław Langiewicz, e-mail: [email protected] dr hab. n. med. Rafał Stec, e-mail: [email protected] dr n. med. Marcin Świerkowski, e-mail: [email protected] lek. Anna Waśko-Grabowska, e-mail: [email protected] dr hab. n. med. Gabriel Wcisło, e-mail: [email protected], tel. 22 681-70-95 lic. Anna Sobocińska Karolina Strychalska Redakcja naukowa: prof. dr hab. n. med. Cezary Szczylik Redakcja pomocnicza: dr inż. Elżbieta Bernatowicz mgr inż. Anna Sikora mgr Aleksandra Klemba

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Sponsorzy i wystawcy:

Główny Partner

Patroni medialni:

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Szanowni Państwo,

W imieniu własnym jak i pozostałych członków Komitetu Naukowego i Komitetu Organizacyjnego mam zaszczyt zaprosić na organizowane przez Fundację Onkologii Doświadczalnej i Klinicznej Międzynarodowe Sympozjum: „Rak nerki, rak prostaty 2015 – Nowe wyzwania”, które odbędzie się w dniach 23-24 października 2015 r. w Hotelu Narvil w Serocku pod Warszawą. Celem sympozjum jest przedstawienie nowych wyników badań klinicznych w zakresie raka nerki oraz raka prostaty. Podczas sympozjum swoje wyniki zaprezentują specjaliści z wiodących ośrodków w kraju i zagranicą. Dodatkowym celem jest stworzenie atmosfery do dyskusji i wymiany poglądów, co przyczyni się do późniejszej współpracy w badaniach naukowych i praktyce klinicznej. Liczę na to, iż zarówno tematyka, jak i ciekawa formuła sympozjum spotka się z zainteresowaniem i spełni Państwa oczekiwania. Serdecznie zachęcam do aktywnego udziału w sympozjum.

W imieniu Komitetu Naukowego i Organizacyjnego Cezary Szczylik

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O FUNDACJI Fundacja powstała 19 kwietnia 1994 r. pod nazwą Fundacja Onkologii Doświadczalnej i Klinicznej. „Celem Fundacji jest wspieranie rozwoju nowoczesnej profilaktyki, diagnostyki i leczenia chorób nowotworowych w oparciu o osiągnięcia światowej biologii, medycyny i psychologii” — ze statutu Fundacji, przyjętego przez Sąd Rejonowy dla m.st. Warszawy, Wydział XVI Gospodarczy Rejestrowy, 12 maja 1994 r. Fundacja realizuje swoje cele, między innymi:  finansując wyjazdy lekarzy i naukowców na praktyki, staże, sympozja, konferencje i zjazdy naukowe,  finansując działania mające na celu upowszechnienie wiedzy o chorobach nowotworowych (kursy, szkolenia, wykłady, publikacje) wśród personelu medycznego,  przyznając środki finansowe i rzeczowe instytucjom medycznym zajmującym się onkologią kliniczną na poprawę stanu profilaktyki, diagnostyki i leczenia,  przyznając środki finansowe i rzeczowe instytucjom naukowym na badania w zakresie poznania istoty, profilaktyki, diagnostyki i leczenia chorób nowotworowych /onkologia doświadczalna/ oraz na wdrażanie nowych metod profilaktycznych, diagnostycznych i terapeutycznych,  upowszechniając w środowisku medycznym i naukowym oraz w społeczeństwie osiągnięć nauk biologiczno-medycznych oraz psychologii w zakresie poznania chorób nowotworowych,  przyznając nagrody i wyróżnienia oraz fundując stypendia za najwybitniejsze osiągnięcia w profilaktyce; diagnostyce i wdrażaniu nowoczesnych terapii w zakresie onkologii w Polsce. Celem Fundacji Onkologii Doświadczalnej i Klinicznej jest zbliżenie nauki z praktyką kliniczną, przyśpieszenie wdrażania nowych metod diagnostyki i leczenia nowotworów i rozwój oryginalnych polskich.

Założona przez nas Fundacja jest wynikiem niecierpliwości, niecierpliwości chorych, lekarzy, naukowców oczekujących przełomu w onkologii, nie tylko zrozumienia istoty choroby, ale przede wszystkim przełamania bezsilności wobec jej niepohamowanego wzrostu. Wiemy również, iż świadomość potrzeby rozwoju nauk podstawowych dla potrzeb medycyny musi być zrozumiała i powszechna, gdyż to właśnie one decydują o postępie. Cezary Szczylik

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SYMPOSIUM’S PROGRAM / PROGRAM SYMPOZJUM

Friday / Piątek 23.10.2015 r. 9.00 – 9.40

Registration of participants / Rejestracja uczestników

9.40 – 9.50

Welcome address / Powitanie uczestników — Cezary Szczylik

Session I: Kidney cancer I / Sesja I: Rak nerki I Session moderators / Moderatorzy sesji: Cezary Szczylik 9.50 – 10.20

Optimal treatment sequence for mRCC / Optymalna sekwencja leczenia dla mRCC – Enrique Grande

10.20 – 10.50 NFZ mRCC TKI treated patients polish database – Final Report / Baza danych NFZ chorych na mRCC leczonych inhibitorami TKI – Marcin Świerkowski 10.50 – 11.20 Hypoxia action on tumor microenvironment: participation to resistance to treatment / Wpływ hipoksji na mikrośrodowisko guza nowotworowego: udział w generowaniu odporności na leczenie – Claudine Kieda 11.20 – 11.35 Coffee break / Przerwa kawowa 11.35 – 12.05 Treating metastatic RCC today – means sequencing / Obecne leczenie mRCC - rola leczenia sekwencyjnego – Camillo Porta 12.05 – 12.35 Renal cell carcinoma – cancer stem cells – do they exist? / Komórki macierzyste raka nerki – istnieją czy nie istnieją? – Anna M. Czarnecka 12.35 – 13.05 Role of hypoxia in shaping cell-mediated cytotoxicity: From immune resistance to immune supression / Rola hipoksji w “kształtowaniu” cytotoksyczności: od immuno-odporności do immunosupresji – Salem Chouaib 13.05 – 13.25 Discussion / Dyskusja 13.25 – 14.25 Lunch / Obiad

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Session II: Kidney cancer II / Sesja II: Rak nerki II Session moderators / Moderatorzy sesji: Gabriel Wcisło, Camillo Porta 14.25 – 14.45 Transmembrane protein genes (TMEMs) and their role in the development of ccRCC / Geny kodujące białka transmembranowe (TMEMs) i ich udział w rozwoju ccRCC – Joanna Wesoły 14.45 – 15.05 First line treatment of metastatic renal cel carcinoma (mccRCC) / Pierwsza linia leczenia zaawansowanego raka nerki – Cezary Szczylik 15.05 – 15.25 Long term survivors after first line sunitinib treated mRCC patients. Polish NFZ database analysis / Chorzy długo żyjący z mRCC po leczeniu pierwszą linią w oparciu o sunitynib. Dane w oparciu o analizę materiałów NFZ – Cezary Szczylik 15.25 – 15.45 TKI-everolimus sequence – PFS and OS. Data from polish NFZ database / Leczenie sekwencyjne TKI-Everolimus – PFS i Os. Analiza danych z programu terapeutycznego leczenia raka nerki NFZ – Andrzej Śliwczyński 15.45 – 16.05 The role of mRCC immunotherapy in targeted agents age / Rola immunoterapii w dobie leków celowanych – Przemysław Langiewicz 16.05 – 16.15 Prognostic factors for overall survival in patients with metastatic renal cell carcinoma treated with first-line tyrosine-kinase inhibitors / Czynniki prognostyczne dla przeżycia całkowitego u pacjentów z rozsianym rakiem nerwokomórkowym leczonych w pierwszej linii inhibitorami kinaz tyrozynowych – Paweł Chrom 16.15 – 16.35 Discussion / Dyskusja 16.35 – 17.00 Coffee break / Przerwa kawowa 20.00

Gala dinner / Uroczysta kolacja

Saturday / Sobota 24.10.2015 r. Session III: Prostate cancer I / Sesja III: Rak prostaty I Session moderators / Moderatorzy sesji: Cezary Szczylik, Piotrz Radziszewski, Iwona Skoneczna 09.00 – 09.20 Descriptive models for cancer treatment in uro-oncology. Results for Poland / Deskryptywne modele leczenia nowotworów urologicznych – wyniki dla Polski – Barbara Więckowska 09.20 – 09.40 Active surveillance (AS) in my everyday practice / Baczna obserwacja w mojej codzienne praktyce praktyce – Andrzej Borkowski

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09.40 – 10.00 Brachyterapy in the treatment of locally advanced prostate cancer / Brachyterapia jako metoda leczenia miejscowo zaawansowanego raka gruczołu krokowego – Anna Kulik 10.00 – 10.15 Coffee break / Przerwa kawowa 10.15 – 10.35 Current radiation oncology in the treatment of locally advanced and oligometastatic prostate cancer / Nowoczesna radioterapia w leczeniu miejscowo zaawansowanego i oligometastatycznego raka stercza – Sergiusz Nawrocki 10.35 – 10.55 What`s going on in radical prostatectomy? / Czy coś nowego w prostatektomii radykalnej? – Tomasz Borkowski 10.55 – 11.15 Life saving prostatectomy / Ratunkowa prostatektomia – Wojciech Rogowski 11.15 – 11.35 State of the art prostate biopsy / Biopsja gruczołu krokowego: nowe techniki – Bob Djavan 11.35 – 11.50 Coffee break / Przerwa kawowa

Session IV: Prostate cancer II / Sesja IV: Rak prostaty II Session moderators / Moderatorzy sesji: Andrzej Borkowski 11.50 – 12.10 Novel systemic therapy of prostate cancer and how they are able to improve results? / Nowe zastosowania terapii systemowych w leczeniu raka stercza- czy mogą poprawić wyniki leczenia? – Iwona Skoneczna 12.10 – 12.30 Are we ready for personalized therapy for prostate cancer?/ Czy jesteśmy gotowi na personalizację leczenia w raku stercza? – Piotr Radziszewski 12.30 – 12.50 Functional complications after radical prostatectomy / Powikłania czynnościowe prostatektomii radykalnej – Piotr Dobroński 12.50 – 13.10 Proctologic complications in patients with prostate carcinoma / Powikłania proktologiczne u pacjentów leczonych z powodu raka prostaty – Małgorzata Kołodziejczak 13.10 – 14.00 Debate: high-risk prostate cancer and how to optimize its treatment / Debata: rak stercza wysokiego ryzyka – optymalizacja leczenia Moderators / Moderatorzy: Iwona Skoneczna, Piotr Radziszewski. Debate participants / Uczestnicy Panelu: Piotr Dobroński, Andrzej Borkowski, Wojciech Rogowski, Tomasz Borkowski Anna Kulik, Sergiusz Nawrocki 14.00 – 15.00 Lunch / Obiad 14.00 – 15.00 Poster session / Sesja posterowa

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Session V: Molecular aspects in kidney cancer and prostate cancer treatments Sesja V: Molekularne aspekty w leczeniu raka nerki i raka prostaty Session moderators / Moderatorzy sesji: Anna M. Czarnecka, John Petros 15.00 – 15.30 Metastatic renal cell carcinoma through the eyes of the international mRCC database consortium / mRCC widziany oczyma the International mRCC Database Consortium – Daniel Heng 15.30 – 16.00 MtDNA mutations and prostate cancer bone metastases / Mutacje mitochondrialnego DNA a przerzuty do kości raka prostaty – John Petros 16.00 – 16.30 Role of oncogenic autophagy in clear cell renal cell carcinoma / Rola autofagii w raku jasnokomórkowym nerki – Maria Czyżyk-Krzeska 16.30 – 16.50 Hypoxia alleviation: new treatments to normalize stably tumor vessels as they adjuvant therapeutic means / Osłabienie hipoksji jako nowy sposób leczenia, normalizujący naczynia krwionośne, w terapii adiuwantowej – Claudine Kieda 16.50 – 17.10 What thyroid hormones have to do with renal cancer? / Hormony tarczycy a rak nerki – Agnieszka Piekiełko-Witkowska 17.10

Conference closing / Zakończenie konferencji

17.15 – 17.30 Coffee break / Przerwa kawowa

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Friday / Piątek 23.10.2015

Session I: Kidney cancer I Sesja I: Rak nerki I Session moderators / Moderatorzy sesji: Cezary Szczylik

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Optimal treatment sequence for mRCC Optymalna sekwencja leczenia dla mRCC Enrique Grande Medical Oncology Department, Ramón y Cajal University Hospital, Madrid. Spain

Since 2005, a large number of different systemic targeted agents have been approved for the treatment of metastatic renal cell carcinoma (mRCC), without any relevant clinic or molecular biomarker that may help us to choose the most optimal first-line and second-line sequence. In first line, sunitinib remains the reference standard of care worldwide in the first line setting and new trials with novel checkpoint inhibitors are all taking sunitinib as the comparator arm. Less optimal alternatives include pazopanib, bevacizumab plus interferon and temsirolimus in patients with poor expected tolerability to sunitinib, intravenous need for administration or poor-risk features. In second line, axitinib data from the AXIS trial are substantially more robust given the inclusion of more patients considered as true second-line, and validly justify the choice of axitinib over everolimus. Nonetheless, the RECORD-1 trial of everolimus may be considered as reference for use as third-line or subsequent lines of therapy. The optimal therapy management of the different alternatives that we have and the subsequent use of these agents are clearly impacting in the clinical benefit and overall survival of our patients with disseminated RCC.

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NFZ mRCC TKI treated patients, Polish database – Final Report Baza danych NFZ chorych na mRCC leczonych inhibitorami TKI Marcin Świerkowski1, Andrzej Śliwczyński2,3, Tomasz Czeleko2, Zbigniew Teter4, Gabriel Wcisło1, Anna M. Czarnecka1, Aneta Ptak-Chmielewska5, Daniel Heng6, Cezary Szczylik1 1

Department of Oncology, Military Institute of Medicine, Warsaw, Poland, 2Department of Drugs Management, National Health Fund Center, Warsaw, Poland; 3Public Health Division, Department of Health Science, Medical University of Lodz, Poland, 4National Health Fund Center, Warsaw, Poland, 5Institute of Statistics and Demography, Warsaw School of Economics, Poland; 6Department of Medical Oncology, Tom Baker Cancer Center, University of Calgary, Calgary, Canada.

Systemic treatment in patients with clear-cell metastatic renal cell cancer (cmRCC) has been changed from cytokine-based therapy to drugs targeting angiogenesis and mammalian target of rapamycin pathway for the last decade. In Poland, tyrosine kinases inhibitors (TKIs) are available as first-line (Sunitinib, Pazopanib) and second-line (Axitinib, Sorafenib) therapy for cmRCC and are reimbursed from public health insurance. National Health Fund (Narodowy Fundusz Zdrowia - NFZ) is the only state institution that reimburses targeted therapies in Poland. Since 2009 patients with cmRCC have been treated with TKIs in one of the Therapeutic Health Care Programs (THCP) established and monitored by NFZ. The primary objective of the THCPs is to prolong overall survival and progression free survival of cmRCC patients by providing the target therapies for the patients. There are strictly defined inclusion, exclusion criteria in the THCP and uniform scheme of efficacy assessment and safety reporting for all patients participating in the programs. We retrospectively analyzed the efficacy and safety of sunitinib, pazopanib, axitinib and sorafenib in cmRCC patients enrolled in THCPs. Results of these analyses will be presented at the symposium.

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Hypoxia action on tumor microenvironment: participation to resistance to treatment Wpływ hipoksji na mikrośrodowisko guza nowotworowego: udział w generowaniu odporności na leczenie Bouchra El Hafny-Rahbi1, Guillaume Collet1, Anna Tejchman1, 2, Krzysztof Klimkiewicz1,3, Claudine Kieda1 Centre de Biophysique Moléculaire, UPR 4301 CNRS, Rue Charles Sadron, 45071 Orléans CEDEX2, France; 2Glycobiology and intercellular interaction team, Institute of Immunology and Experimental Therapy, PAN, Ul. Rudolfa Weigla 12, 53-114 Wroclaw, Poland; 3Jagiellonian University, Faculty of Biochemistry, Biophysics and Biotechnology, Ul. Gronostajowa 7, 30-387 Kraków, Poland. 1

Renal cell carcinoma is particularly angiogenic and resistant to treatments. It is largely treated by antiangiogenic therapies. Tumor response to targeted agents is often heterogeneous and most responsive patients develop resistance. Moreover in RCC as in other type of cancer in development, the microenvironment is such that infiltrating cells from the immune system as well as non-immune cells as fibroblasts, participate to tumor development. RCC immunogenicity and angiogenicity combine to make it resistant and metastatic. RCC is immunogenic, infiltrated with immune cells, including macrophages and T lymphocytes. But, in this case tumor-infiltrating lymphocytes (TIL) counts correlate with poorer prognosis. Mechanism and pathophysiologic significance of TILs inactivation remains unclear and related to immune checkpoints, regulatory T cells (Treg), and myeloid derived suppressor cells (MDSC) suppressing T-cell responses. Thus, in RCC immunosuppression is preponderant over efficient immune response. This conditions microenvironmentlinked reactions thus treatment efficacy. Angiogenesis is hypoxia-conditioned, which is responsible for its pathological features sustaining hypoxia and above-mentioned deleterious mechanisms of aggressiveness. Carreau, A., et al. J. Cell. Mol. Med. 15, 1239-1253, (2011).Collet, G. et al. Vasc. Pharmacol.56, 252-261,(2012).Collet, G. et al. Cancer Ther. 13, 165-178, (2014). Kieda, C. et al. J. Mol. Med. 91, 883-899, (2013).

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Treating metastatic RCC today means sequencing Obecne leczenie mRCC – rola leczenia sekwencyjnego Camillo Porta1,2, Ilaria Toscani1 1

Medical Oncology, I.R.C.C.S. San Matteo University Hospital Foundation, Pavia, Italy; 2Italian Group of Nephro-Oncology/Gruppo Italiano di Oncologia Nefrologica (G.I.O.N.)

The treatment of mRCC has dramatically improved since the development of targeted agents, directed against either the VEGF pathway, or the serine-threonine kinase mTOR. Due to the present availability of 7 registered active agents, the natural history has changed for the best, even though a cure is still out of sight. Consequent to this abundance of active agents, more and more patients are, and will be, treated beyond first-line. This also means that sequencing is key for maximizing benefit to mRCC patients. To date, treatment choice in second- and third-line is still largely empirical, based on few randomized controlled trials, and other studies, prospective and retrospective, often highly biased. At present, with regard to choosing the second line treatment after the failure of therapy with VEGFR-TKIs, the continued inhibition of the VEGF/VEGR pathway, or else the switch to an mTOR inhibitor, is recommended. These two options are characterized by partly different targets, completely different toxicity profiles, but a comparable efficacy. As regards third line treatment, where a sequence of two VEGFR-TKIs has been used beforehand, the choice is represented by the mTOR inhibitor everolimus, whilst if a VEGFR-TKI followed by everolimus has been chosen, a return to VEGF pathway inhibition is suggested. Notably, we should not neglect the fact that in treatment lines beyond the first, when the expected survival benefit cannot be huge, toxicity and comorbidity concerns should be regarded as major determinants of the therapeutic choice. From a practical viewpoint, treatment choices should be aimed at trying to match each drug’s profile (in terms of mechanism of action, efficacy results achieved, and especially safety profile) with each patient’s characteristics and needs, taking into account that the overall quality of care, not just the single agent chosen, makes the differences. We badly need biomarkers to guide our clinical decisions, and every effort should be made to find putative ones, and then validate and translate them into clinical practice. In this regard, recent data suggesting that mutations in mTOR pathway-related genes may predict long-term benefit from mTOR inhibitor therapy is of the utmost importance, even though these preliminary data still need to be prospectively validated on an adequate sample size. Likely, in the next 2 to 3 years the scenario will dramatically change with the registration of cabozantinib and nivolumab, given the announced positive results of their pivotal phase III trials. Future potential changes in the first-line standard of care could completely turn this scenario upside-down again in 3 to 5 years. Safety will continue to represent an extremely important issue, expecially in treatment lines where the expected survival

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benefit is limited, whilst quality of life retains its huge importance. The so-called law of diminishing returns may no longer apply to the mRCC field, in light of the above novelties and of the increasing activity of novel agents. Also for novel agents (cabozantinib, nivolumab and others which will realistically come), the quest for biomarkers will be crucial. Will MET status or PD1/PDL1 expression help us to select patients more likely to respond to these agents? Finally, economical considerations will play more and more a key role, limiting the availability of agents in many countries around the world, not necessarily in low income ones.

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Renal cell carcinoma – cancer stem cells – do they exist? Komórki macierzyste raka nerki – istnieją czy nie istnieją? Anna M. Czarnecka1, Mohammed I. Khan1, Zofia F. Bielecka1,2, Damian Matak1,2, Wojciech Solarek1,2, Anna Kornakiewicz1,2, Cezary Szczylik1 1

Laboratory of Molecular Oncology, Department of Oncology, Military Institute of Medicine, Warsaw; School of Molecular Medicine, Medical University of Warsaw, Warsaw, Poland

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Cancer is initiated by unique population of cells known as cancer stem cells (CSCs) or tumor initiating cells (TIC). CSCs are responsible not only for tumor development but also for disease progression, disease recurrence, aggressiveness, metastatic spread, and finally chemo- and radio-therapy resistance. The existence of CSCs has been proven in different cancer types. We aim to confirm the presence of cancer including renal cell cancer. It was previously shown that RCC-TICs candidates express of a wide array of cell surface markers such as CD105, CD133, ALDH1, CD44, or CXCR4. We aim to investigate presence of CD105/133 subpopulations in RCC in vitro model and further characterize these cells. We have shown that CD105 cells represent as much as 5.9±0.9% of the total tumor cell numbers in RCC. The presence of CD105/CD133 subpopulations was measured in primary and metastatic RCC cell lines. RCC-CSC candidates were screened for established mesenchymal and cancer stem cell markers including: CD146, CD90, CD73, CD45, CD44, CD34, CD24, CD19, CD11b, HLA-DR and alkaline phosphatase. Activity of tyrosine kinase inhibitors (sunitinib, sorafenib, axitinib) against RCC-CSCs was measured. Gene expression profile of CD105 cells isolated form primary and metastatic tumors was compared. Proteome of tyrosine kinase inhibitor resistant cells was investigated.

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Role of hypoxia in shaping cell-mediated cytotoxicity: from immune resistance to immune suppression Rola hipoksji w „kształtowaniu” cytotoksyczności: od immuno-odporności do immunosupresji Salem Chouaib, Messai Yosra, Noman Zaeem, Bernard Escudier INSERM-U1186, Institut Gustave Roussy, Villejuif-France

Cytotoxic T lymphocytes (CTL) and NK cells are important effector cells in tumor rejection and play a crucial role in host defense against malignancies. However, these cells are engaged in an active and complex molecular cross-talk with tumor stroma components including hypoxia that has serious implications for immunological recognition of tumor cells in shaping stroma reactivity. We obtained data indicating that hypoxic stress-induced tumor target adaptation, compromises the effectiveness of killer cell activity through a multiparametric process conferring a resistant phenotype. We provided in particular evidence indicating a critical role of HIF-2a in the regulation of Renal cell carcinoma to NK-mediated cell lysis. We will illustrate how hypoxic stress is associated with the emergence of immune suppression and the regulation of some critical immune check-points molecule ligands (PDL1). How hypoxia, a major hallmark of most solid tumors, in the tumor microenvironment plays a central role in the evolution of immune escape mechanisms by tumor cells will be discussed.

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Friday / Piątek 23.10.2015

Session II: Kidney cancer II Sesja II: Rak nerki II Session moderators / Moderatorzy sesji: Gabriel Wcisło, Camillo Porta

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Transmembrane protein genes (TMEMs) and their role in the development of ccRCC Geny kodujące białka transmembranowe (TMEMs) i ich udział w rozwoju ccRCC Tomasz Wrzesiński, Joanna Wesoły Laboratory of High Throughput Technologies, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, Poznań

VHL inactivation is the most established molecular characteristic of clear cell renal cell carcinoma (ccRCC), with only a few additional genes implicated in development of this kidney tumor. In recently published ccRCC gene expression meta-analysis study we identified a number of deregulated genes with limited information available concerning their biological role, represented by gene transcripts belonging to transmembrane proteins family (TMEMs). TMEMs belong to a heterogeneous, poorly characterized group of predicted in silico membrane-integral proteins with limited sequence homology between the family members and comprise of approximately 300 members. Although TMEM expression data in many cancers is available in public repositories, only a few TMEMs were subjected to detailed functional analysis, and those do not include TMEMs deregulated in ccRCC. The mechanism of deregulation and the function of majority of ccRCC deregulated TMEMs remains unclear, including their contribution to the development and progression of ccRCC. Recently we confirmed the deregulation of 10 TMEMs in an independent study of 76 ccRCC tumors and associated the changes in TMEM expression in tumors with the clinical outcome. Additionally we performed bioinformatics predictions of TMEM topology, function and localization and using genomic technologies we attempted to understand the mechanisms underlying TMEM deregulation in ccRCC.

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First line treatment of metastatic renal cell carcinoma (mccRCC) Pierwsza linia leczenia zaawansowanego raka nerki Cezary Szczylik Department of Oncology, Military Institute of Medicine, Warsaw, Poland

Renal cell carcinoma accounts for 2-3% of adult malignancies and represents the seventh most common cancer in men and the ninth among women. In Poland more than 5000 cases of RCC occurred per year. The incidents of all stages of RCC has increased over the past several years. For medical oncologist the group of mccRCC remains the most challenging. Since 2007 first active substances have been described and tested in phase III clinical trials, but the questions which of those novel drugs should be given as a first line remains a question. Based on polish national health care (NFZ) database we are presenting pooled data of all polish patients with mccRCC given TKI in a first line.

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Long term survivors after first line sunitinib treated mRCC patients. Polish NFZ database analysis Chorzy długo żyjący z mRCC po leczeniu pierwszą linią w oparciu o sunitynib. Dane w oparciu o analizę materiałów NFZ Cezary Szczylik Department of Oncology, Military Institute of Medicine, Warsaw, Poland

Among mccRCCC patients a subset achieves long-term responses with TKI therapy. We are presenting a retrospective analysis from National Health Care (NFZ) database which includes patients who responded with complete responses, partial remissions and stable disease for more than 36 months. Sunitinib treated patients are the largest in polish database and data of this group contained same criteria usually used in phase III clinical trials. Sunitinib achieves long term responses in more than 20% subset of polish patients. Treatment of a such big population for a long time raises many important questions – how long we need to treat this subset of patients?, is it possible to stop therapy after achieving such long response and then observe? and then in case of disease recurrence restart sunitinib? These questions we address to health care providers offering a strict cooperation.

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TKI-Everolimus sequence – PFS and OS. Data from Polish NFZ database Leczenie sekwencyjne TKI-Everolismus – PFS i OS. Analiza danych z programu terapeutycznego leczenia raka nerki NFZ Andrzej Śliwczyński1,2, Marcin Świerkowski3, Tomasz Czeleko1, Zbigniew Teter4, Gabriel Wcisło3, Anna M. Czarnecka3, Aneta Ptak-Chmielewska5, Daniel Heng6, Cezary Szczylik1 Departament Analiz i Strategii, Centrala Narodowego Funduszu Zdrowia w Warszawie; 2Oddział Zdrowia Publicznego, Wydział Nauk o Zdrowiu, Uniwersytet Medyczny w Łodzi; 3Klinika Onkologii, Wojskowy Instytut Medyczny w Warszawie; 4Centrala Narodowego Funduszu Zdrowia w Warszawie; 5 Instytut Statystyki i Demografii Szkoły Głównej Handlowej w Warszawie; 6 Department of Medical Oncology, Tom Baker Cancer Center, University of Calgary, Calgary, Canada 1

There are limited published data on the clinical outcome of clear-cell metastatic renal cell cancer (cmRCC) patients treated with TKIs-Everolimus sequence in real-world clinical practice at a national level. In Poland, treatment with the target agents is currently available and monitored in the Therapeutic Health Care Programs (THCPs) established by the state institution – National Health Fund (NFZ). The primary aim of the THCPs is to prolong overall survival and progression free survival of cmRCC patients by providing the target therapies for the patients. Inclusion, exclusion criteria and the scheme of toxicity and efficacy assessment are defined in each the THCP. About sixty health care providers, which enroll patients in the THCPs, collect the hard copies of patients’ medical records for local NFZ monitoring which constitutes the fundamental method of the program auditing. Some defined patients’ medical records are entered electronically into the central NFZ databases from the beginning of treatment and then every few weeks. Based on the central NFZ databases we performed retrospective analysis of efficacy of the TKIs-Everolimus sequence in cmRCC patients. The results of the analysis will be presented at the symposium.

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The role of mRCC immunotherapy in targeted agents age Rola immunoterapii w dobie leków celowanych Przemysław Langiewicz Department of Oncology, Military Institute of Medicine, Warsaw, Poland

Non-specific immunotherapy was the first effective therapeutic option introduced for the mRCC treatment in the late 70s. Efficacy and toxicity profile of both INF alfa alone and combined with IL-2, chemotherapeutic agents or hormones were analyzed in many prospective clinical studies. The review by R.J. Motzer, which include the analysis of several phase III clinical trials (from 1982 to 1996), has revealed that pooled median OS and median TTP for immunotherapy were 13 and 4,7 months, respectively. Treatment optimization require adequate mRCC patients selection based on the MSKCC scoring system. Efficacy of immunotherapy was confirmed in AVOREN study (2004-2008 OS analysis), in which IFN alfa-2a combined with Bevacizumab treatment resulted in 10,2 months of median PFS and 23,3 months of median OS, compared with 5,4 and 21,3 months for IFN alone. Currently immunotherapy is focused on inhibition of PD-1 receptor (Nivolumab) and CTLA-4 pathway (Ipilimumab), what leads to the activation of anticancer Tlymphocyte response. Those new agents are under study in mRCC therapy used both alone and in combination with TKI or mTOR inhibitors.

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Prognostic factors for overall survival in patients with metastatic renal cell carcinoma treated with first-line tyrosine kinase inhibitirs Czynniki prognostyczne dla przeżycia całkowitego u pacjentów z rozsianym rakiem nerkowokomórkowym leczonych w pierwszej linii inhibitorami kinaz tyrozynowych Paweł Chrom, Rafał Stec, Cezary Szczylik, Aleksandra Semeniuk-Wojtaś, Lubomir Bodnar Department of Oncology, Military Institute of Medicine, Warsaw, Poland

Background. The study investigated which demographical, clinical and pathological features may have a prognostic role for overall survival (OS) in patients with metastatic renal cell carcinoma (mRCC) treated with first-line tyrosine-kinase inhibitors (TKI). Methods. Patients with mRCC, who started treatment with first-line TKI between January 2010 and December 2014 in Department of Oncology, Military Institute of Medicine, Warsaw, Poland, were included. Cox proportional hazards regression was used to find independent factors for OS. Two bootstrap procedures with calculation of bias-corrected concordance index (c-index) were used to validate computed model. Results. Overall, 266 patients were included. Median OS for the whole cohort was 24.8 months. Six factors were found to be independently associated with poor survival: Eastern Cooperative Oncology Group performance status > 0 (p < .0001), Fuhrman grade 3 to 4 (p < .0001), hemoglobin < lower limit of normal (p < .0001), lactate dehydrogenase > upper limit of normal (p = .0011), neutrophil-to-lymphocyte ratio (NLR) ≥ 4 (p < .0001) and > 2 metastatic sites (p = .0012). Bias-corrected c-index was .751. Conclusion. In addition to factors previously reported in the literature, Fuhrman grade and NLR independently impacted survival. Despite satisfactory prognostic ability of the model, it needs to be validated externally using larger dataset.

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Saturday / Sobota 24.10.2015

Session III: Prostate Cancer I Sesja III: Rak prostaty I Session moderators / Moderatorzy sesji: Cezary Szczylik, Piotr Radziszewski, Iwona Skoneczna

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Descriptive models for cancer treatment in uro-oncology. Results for Poland Deskryptywne modele leczenia nowotworów urologicznych – wyniki dla Polski Barbara Więckowska

Departament Analiz i Strategii, Ministerstwo Zdrowia; Katedra Ubezpieczenia Społecznego, Szkoła Główna Handlowa

Descriptive models for cancer treatment are widely used in different countries in the decision-making process regarding plans of health benefits that would satisfy needs of population in the area of oncological treatment. Ministry of Health with the team of experts, including oncologists, econometricians, statisticians and IT specialists made an attempt to create such models for different 25 groups of cancers, from which 3 were dedicated to urology. The present study aims to show the results of descriptive models of urological cancers including bladder cancer (C67 according to ICD-10 classification), prostate cancer (C61) and kidney cancer (C64, C65, C66) in the field of surgery, chemotherapy and radiotherapy. On the basis of data from National Cancer Register (KRN) and National Health Fund the estimation of cancer incidence in Poland was provided. Also the distribution of treatment patters was presented - such data could may be a valuable source of information concerning the standards of oncological patients’ treatment in Poland and provide the basis for identifying areas of health care needed to be improved in the nearest future.

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Active surveillance (AS) in my everyday practice Baczna obserwacja w mojej codziennej praktyce Andrzej Borkowski Urological Department Warsaw Medical University

That is urgent need for a treatment option to manage the high proportion of prostate cancer (PC) with clinically low risk of progression. Active surveillance (AS) has been proposed with regard to this aim. There is no consensus on the optimal method of selecting men for AS with traditional markers (different protocols) and the role of new imaging modalities and new histologic, proteomic or genetic markers. Only limited data (4-7 years of follow up) are currently available. The longest observation from Toronto show that 15 years progression to metastatic disease was 2.5 % at 15 years. In recently published meta-analysis of 26 longest series (Simpkin AJ, Eur Urol 2015, 67: 993-1025) mean follow up is no longer than 3.5 years. We know that in most low risk tumors progression occur after 10-15 or even 20 years. So for a time being we have not strong arguments for a long term safety of AS programs. On the other hand AS allow to avoid overtreatment and the complications of treatment in many men with low risk disease and results in a decrease in number needed to treat. Taken together into the consideration I do not propose AS for healthy and younger men. The exception of the rule is conscious decision of well-informed patient to postpone for some years (not to avoid) the active treatment due to some important reason. I consider AS relatively save only in well selected men with low risk PC and a high risk of medium term competing mortality or life-expectancy < 10 y. Patient decided to follow AS should sign special “agreement form” where all disputable problems are presented.

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Brachytherapy in the treatment of locally advanced prostate cancer Brachyterapia jako metoda leczenia miejscowo zaawansowanego raka gruczołu krokowego Anna Kulik Zakład Brachyterapii, Centrum Onkologii – Instytut im. Marii Skłodowskiej-Curie w Warszawie

In patients with clinically localized prostate cancer, brachytherapy is commonly used radical treatment options. In that clinical cases brachytherapy as monotherapy (permanent low dose rate - LDR or interstitial high dose rate - HDR), or in combination with EBRT (external beam radiation therapy) are commonly performed. These modalities have shown biochemical and clinical control rates comparable to those in patients after radical prostatectomy. In view of the good oncological results and tolerability of cancer treatment, brachytherapy is increasingly popular method of treatment. According to data from the NFZ, in 2014 the number of BRT patients was higher than radical prostatectomy. However, in the absence of randomized long follow up clinical trials, the optimal therapeutic approach remains controversial. EBRT and BRT dose escalation for organ-confined prostate cancer improves local disease control. So far, published results show that the BRT both alone and in combination with EBRT is well tolerated and with acceptable level of side effects.

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Current radiation oncology in the treatment of locally advanced and oligometastatic prostate cancer Nowoczesna radioterapia w leczeniu miejscowo zaawansowanego i oligometastatycznego raka stercza Sergiusz Nawrocki Katedra Onkologii i Radioterapii, Śląski Uniwersytet Medyczny w Katowicach

Wskazania do radioterapii raka gruczołu krokowego są obecnie bardzo szerokie. Radioterapia jest równoważną do radykalnej prostatektomii metodą leczenia wczesnego raka prostaty i leczeniem z wyboru choroby zaawansowanej miejscowo. Radioterapia jest obecnie stosowana również po radykalnej prostatektomii u chorych z czynnikami nawrotu miejscowego. Coraz szerzej obok klasycznego frakcjonowania przy pomocy technik IMRT i IGRT stosuje się radykalne hypofrakcjonowanie, które ze względu na postulowaną bardzo niską wartość parametru “alfa/beta” dla komórek raka prostaty pozwala na uzyskanie wyjątkowo korzystnego współczynnika terapeutycznego (wysoka skuteczność/niska toksyczność) przy tym sposobie leczenia. Radykalne hypofrakcjonowanie jest jednocześnie wygodne dla chorych i wiąże się z lepszą jakością życia (w tym funkcjonowaniem w sferze seksualności) w porównaniu do klasycznie frakcjonowanej radioterapii (wyniki własne badania prospektywnego). Wiele kontrowersji budzi zastosowanie protonoterapii w leczeniu raka gruczołu krokowego. Jego wyższość w porównaniu do terapii wiązką fotonową polega na niższej dawce w otaczającej zdrowej tkance; mankamentem natomiast jest wyższe ryzyko tzw. “ błędu geograficznego” i efektywność kosztowa. Ciekawym pomysłem jest (1) radykalna radioterapia chorych z chorobą oligometastatyczną i zastosowanie (2) radioterapii w skojarzeniu z immunomodulacją (odblokowanie tzw. immune checkpoints) w celu uzyskania tzw. efektu abskopalnego. Obie koncepcje są obecnie w fazie ewaluacji w badaniach klinicznych i dają nadzieję na istotny postęp w leczeniu choroby zaawansowanej.

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What`s going on in radical prostatectomy? Czy coś nowego w prostatektomii radykalnej? Tomasz Borkowski

Department of Urology, Medical University of Warsaw, Poland

Prostatektomia radykalna w leczeniu raka gruczołu krokowego wykonywania jest od 1905 roku, jednakże dopiero ostatnie 20 lat przyniosło znaczące zmiany zarówno jeśli chodzi o wskazania, technikę operacyjną a także wyniki zarówno onkologiczne jak i czynnościowe. W ostatnich latach odnotowuje się tendencję do ograniczenia wskazań do leczenia operacyjnego pacjentów z rakiem bardzo niskiego i niskiego ryzyka na korzyść aktywnej obserwacji a coraz więcej wskazania do leczenia operacyjnego znajduje się wśród chorych na raka stercza wysokiego ryzyka, co do tej pory było domeną radio-hormonoterapii. Mimo, iż prospektywne badania z randomizacją porównujące radioterapię z leczeniem operacyjnym się toczą, to jednak mamy coraz więcej pośrednich dowodów na to, że leczenie zabiegowe jest równie skuteczne jak naświetlania. Co więcej, okazuje się, że usunięcie prostaty znacząco poprawia przeżycie ogólne u chorych z zajętymi węzłami chłonnymi w porównaniu do grupy pacjentów, u których takiego leczenia zaniechano. Pojawiają się pierwsze doniesienia o zasadności zastosowania prostatektomii radykalnej nawet u pacjentów z chorobą rozsianą (tzw. przerzuty oligometastayczne – czyli nie więcej niż 3), jednakże takie postępowanie nie jest jeszcze rekomendowanie przez Europejskie ani Amerykańskie Towarzystwo Urologiczne do czasu ukończenia stosowych badań kliniczny.Od kilku lat obserwuje się też poprawę wyników czynnościowych, ograniczenie liczby powikłań okołooperacyjnych i odległych oraz burzliwy rozwój technik mało inwazyjnych.Pierwsze doświadczenia z prostatektomią radykalną z użyciem robota (RAPL – robot assisted radical prostatectomy) pochodzą z 2000 roku natomiast w 2015 roku w USA ponad 95% pacjentów zdecydowanych na leczenie operacyjne wybiera taką formę zabiegu (a nie klasyczną operację otwarta). RARP wiąże się z minimalną utratą krwi, krótkim czasem hospitalizacji oraz ograniczeniem powikłań czynnościowych, głownie pod postacią wcześniejszej ustąpieniem lub braku zaburzeń wzwodu. W samej technice zabiegu testuje się nowe możliwości wizualizacji 3D oraz wykorzystania tzw augmented realty (rozszerzona rzeczywistość) jak wykonywanie zabiegu całkowicie podpowięziowo z

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zaoszczędzeniem kompleksu więzadeł łonowo-pęcherzowych i powięzi miedniczej czy też dostęp przezotrzewnowy z zaoszczędzeniem przestrzeni Retziusa. Mimo, że od wielu lat wieszczy się szybki koniec leczenia operacyjnego raka gruczołu krokowego, to jednakże dziś nic nie wskazuje na to aby w najbliższych latach prostatektomia radykalna miała zostać zapomniana a wiele wskazuje na to, że zabieg ten będzie stale udoskonalany.

Life saving prostatectomy Ratunkowa prostatektomia Wojciech Rogowski

Clinical Department of Urology and Urological Oncology, Central Clinical Hospital of the Ministry of Interior in Warsaw, Poland

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State of the art prostate biopsy Biopsja gruczołu krokowego: nowe techniki Bob Djavan New York University (NYU) and Vienna; Director Vienna Urology Foundation, Vienna, Austria; Chairman Regional Office of the European Association of Urology (EAU); Executive Board Member European Society of Oncological Urology (ESOU)

Prostate biopsies are still today the pillars but also the controversial issue of prostate cancer diagnosis. Random biopsies are far from efficient in detecting all tumors and even less efficient in detecting all significant cancer spots. To improve sensitivities and specificities, increasing the biopsy core numbers, targeting more lateral aspects and encouraging repeat biopsies have been recommended. Recently, histoscanning and template biopsies have been introduced to further improve biopsy quality and efficiency. The latest innovations include the fusion of MRI pictures with the transrectal US image to offer optimal targeting of suspicious areas. And yet, these efforts are far from solving the main problem. How can we perform a biopsy and be confident to detect most of the cancers, ie significant malignant areas. New and old markers such as bPSA, pro PSA, p2PSA among others (1), as well as improved imaging techniques based on fusion of TRUS- and MRIguided images will certainly improve optimal patient selection and lesion targeting and by such improve specificity and reduce the number of men undergoing unnecessary prostate biopsies. However, the entire prostate cancer detection path is handicapped by an ailing TRUS biopsy procedure, which has little evolved since Thomas Stamey’s first report on sextant prostate biopsies. Most of todays research efforts rely on the identification of new and better serum markers for prostate cancer. In general the biopsy procedure and its outcome will judge the statistical performance of an investigated test and This is indeed a major source of concern, since the biopsy procedure itself is not standardized among the studies. Biopsy targets, core-numbers, and 3D planing of the needles may vary. Therefor, the conclusions of these investigations are more closely related to the technical performance of the biopsy procedure itself rather than the performance of the marker being investigated. Indeed Sensitivity of the biopsy procedure itself in detecting cancer did not exceed 33%. We have to acknowledge that the success of TRUS biopsy still relies primarly on sampling, which reflects the fact that increasing the number of cores will increase cancer detection rate and complications. Increasing the biopsy cutting length and depth/number of anterior-directed cores (14-18 cores) improved the detection rate for tumors ≥ 0.5cc in the 12 core scheme. The most optimal feature was the Utilization of a 22 mm cutting length and a 12 core scheme with additional volume-adjusted anteriordirected cores. Using this combination, 100% of ≥ 0.5cc tumors in prostates < 50cc in volume and 94.7% of ≥ 0.5cc tumors in prostates > 50cc in volume were detected. The

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ProtecTA study found significant inter-departmental differences in the use of local anaesthetic, antibiotic prophylaxis, pain levels, UTI related complications, cancer detection and strategies toward repeat biopsies, suggesting that changes to practice could markedly improve outcomes. Noting the flaws and inter-departmental variabilities, regarding prostate biopsies, one other major caveat is obvious: lack of standardization. Complication rate of TRUS guided biopsies are low (4.3% in the current study, and 2.7% in the literature). This is mainly due to the wide range use of antibiotics before, during or after the procedure. Clear recommendations have not been given by the AUA nor the EAU, pending adequate trials. However, Fluoroquinolones (FQ) remain one of the standard drugs in the prophylactic regimen with 97.3% of men remaining infection and sepsis free according to the literature. TRUS biopsy complications are important factors in the current controversy addressing PSA screening and early detection as well as the need for repeat and saturation biopsies. Those rejecting Screening point to the high complication rates to be encountered if screening policies would be implemented and equally question the validity of never-ending repeat biopsies and saturation protocols. These concerns are highlighted by the lack of adequate randomized studies and trials. While we are accumulating an enormous database of prostate biopsies worldwide, valid conclusions and lessons are rare. Although 50% of patients who will harbor urosepsis or febrile UTIs will be diagnosed with FQ-resistent E.coli , more studies are needed to adress the impact of FQ resistance. It is however wrong to use TRUS biopsy-related morbidity as an argument against screening/early detection. Larger randomized trials are still lacking to evaluate optimal regimen, dosage and timing of antibiotic regimen but with a 97% infection free rate today, those studies are not expected in the near future. One of the major caveats remains the individual biopsy planning, number of cores, direction and target identification, which is far from standardized, reflecting the large variation in cancer detection rates observed (rates ranging from 23% (CI 14 to 36) to 53% (CI 40 to 65) across centers).Tools such as The Vienna Nomogramm or prostate cancer risk Calculators (PCPT or SWOP) may help individualise prostate cancer diagnosis. Multiparametric MRI based biopsies and specifically TRUS-mpMRI fusion protocols are the future. In times of Active surveillance(AS) and focal therapy, prostate cancer diagnosis needs to be individualized. The options range between TRUS/MRI fusion biopsies, targeted biopsies to template biopsies and the decision depends on the individual patient and the clinical setting. TRUS mpMRI fusion biopsies could be enhanced by involving 14-18 cores (or more if volume increases), involve the anterior zones of the prostate and employ a 22mm cutting length of the biopsy core versus a 15-17mm core as is used currently. Whether, transperineal biopsies are to be involved or added, still need s to be evaluated. In conclusion, TRUS biopsies have evolved over the past years and specially in the last few years based on significant improvements of imaging techniques. Whereas, MRI was only marginal recently, it has become a core element of prostate cancer diagnosis.The future will see biopsies being based on mpMRI and the procedure itself based on a software fusion of the MRI picture and the TRUS image.

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Saturday / Sobota 24.10.2015

Session IV: Prostate cancer II Sesja IV: Rak prostaty II Session moderators / Moderatorzy sesji: Andrzej Borkowski

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Novel systemic therapy of prostate cancer and how they are able to improve results ? Nowe zastosowania terapii systemowych w leczeniu raka sterczaczy mogą poprawić wyniki leczenia? Iwona Skoneczna Department of Urological Oncology, M. Skłodowska-Curie Oncology Center, Warsaw, Poland

Na przestrzeni ostatniego roku jesteśmy świadkami kolejnej ewolucji leczenia systemowego zaawansowanego raka gruczołu krokowego. Po wprowadzeniu do arsenału nowych leków o potencjale wydłużenia przeżycia w mCRPC (metastatic Castration Resistant Prostate Cancer) takich jak abirateron, enzalutamid, sipuleucelT, kabazytaksel czy Rad 223, przyszedł czas na nowe oblicze starej chemioterapii docetakselem. W oparciu o wyniki badań CHAARTED, STAMPEDE i GETUG 15 w 2015 roku zmienione zostały zalecenia NCCN i ESMO. Aktualnie rekomendują wczesną chemioterapię docetakselem łącznie z hormonoterapią u chorych z zaawansowwanym hormonowrażliwym rakiem prostaty z przerzutami ( mHSPC – metastatic Hormone Sensitive Prostate Cancer), którzy są w dobrym stanie ogólnym i kwalifikują się do chemioterapii. Optymalna kolejność, moment włączenia i długość stosowania poszczególnych leków to prawdziwy State of the Art, który zostanie przedyskutowany.

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Are we ready for personalized therpay for prostate cancer? Czy jesteśmy gotowi na personalizację leczenia w raku stercza? Piotr Radziszewski Department of General, Oncological and Functional Urology, Medical University of Warsaw, Warsaw, Poland

Personalized therapy brings the highest clinical efficacy, reduce side effect, decrease costs. It is the ideal treatment, however it requires markers allowing personalization. Do such markers exist for prostate cancer? Each year our knowledge about prostate cancer biology increases. We identified most common gene mutations responsible for cancer pathways. There are more than 70 single nucleotide polymorphisms (SNPs) reported to be associated with prostate cancer risk. With the implementation of new techniques, including new generation sequencing, there have been created tools which can predict prostate cancer aggressiveness (e.g.Prolaris®, OncoType®, OurView®). Other tests like NADiA® and Decipher® can predict clinical and biochemical relapse after radical treatment. But the real progress represents the identification of androgen receptor splice variant 7 (AR-7) identification in circulating tumor cells. With AR-7 measurement it will be possible to predict the clinical response on enzalutamide or abiraterone therapy. Another example of already introduced to clinical practice personalization is the use of PET/CT imaging enhanced with prostate surface membrane antigen (PSMA), which allows identification of metastatic sides with high accuracy and sensitivity. Answering the question, yes we are ready and we are already implementing the personalized therapy in prostate cancer.

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Functional complications after radical prostatectomy Powikłania czynnościowe prostatektomii radykalnej Piotr Dobroński Department of General, Oncological and Functional Urology, Medical University of Warsaw, Warsaw, Poland

Radical prostatectomy (retropubic), endoscopic (most often laparoscopic extraperitoneal) and robotic (RARP) are the most often performed urologic procedures. Their early and late oncologic outcomes in clinically organ confined cancer are comparable though functional outcomes (continence and erectile function) may differ and depend upon variety of factors. They are related to preoperative sexual and continence function as well as BMI. In the past 5 years data appeared suggesting that surgical technique, mainly type / grade of nerve sparing i.e. the choice of surgical plane (intrafascial vs interfascial) during prostate dissection. There is also data on better continence and sexual function recovery after RARP. New techniques are emerging to visualize and identify neurovascular bundles and cavernous nerve branches e.g. optical coherence tomography, fluorescence induced guided surgery (FIGS) and Doppler US identification of cavernosal arteries. Prediction of long term functional status (2 yrs) upon early results (3-12 months) is feasible. New drugs (angiotensin receptor blockers) and techniques (penile vibratory stimulation) are tested in penile postoperative rehabilitation, besides 5-PDI, ICI, MUSE and vacuum devices. Female partner sexual preop function seems also to play role in patients compliance with postop penile rehabilitation program.

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Proctologic complications in patients with prostate carcinoma Powikłania proktologiczne u pacjentów leczonych z powodu raka prostaty Małgorzata Kołodziejczak1, Tomasz Szopiński2, Iwona Sudoł-Szopińska3,4 Proctology Centre, St. Elizabeth Hospital in Warsaw, Goszczyńskiego 1; 2Department and Clinic of

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Urology, Collegium Medicum Jagiellonian University, Cracow Grzegórzecka 18; 3Department of Radiology Institute of Rheumatology in Warsaw, Spartańska 1; 4The Department of Diagnostic Imaging, Second Faculty of Medicine, Medical University of Warsaw

Prostate carcinoma is the second most common cancer in men in Poland. There has been a significant increase in the number of patients who can undergo radical cancer therapy, i.e. resection of the prostate or radiotherapy, due to the progress in cancer diagnostics.Each of these therapies involves the risk of various complications, including proctologic ones, which results from the fact that prostate and rectal wall are situated in immediate vicinity. Damage to the rectal wall during surgery can lead to the formation of abscess, rectal bladder fistula or rectourethral fistula. Damage to the pelvic neural plexuses can cause the symptoms of stool and gas incontinence. Such functional disorders require conservative management (diet, behavioural training, biofeedback).Radiation can cause damage to the rectal mucosa and the associated bleeding, diarrhoea, frequent rectal tenesmus as well as stool and gas incontinence. Usually conservative therapy using different types of medications. Surgical treatment is only used in severe cases of radiation proctitis. Chemotherapy can cause diarrhoea and, consequently, rectal irritation and pruritus, which require symptomatic treatment. Patients with prostate carcinoma can also develop proctologic conditions which are independent of cancer. The method of treatment of these diseases depends on the overall condition and the immune status of the patient.

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Saturday / Sobota 24.10.2015

Session V: Molecular aspects in kidney cancer and prostate cancer treatments Sesja V: Molekularne aspekty w leczeniu raka nerki i raka prostaty Session moderators / Moderatorzy sesji: Anna M. Czarnecka, John Petros

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Metastatic renal cell carcinoma through the eyes of the international mRCC database consortium mRCC widziany oczyma The International mRCC Database Consortium Daniel Heng Department of Medical Oncology, Tom Baker Cancer Center, University of Calgary, Calgary, Canada

This talk will highlight some of the population-based analyses performed by the IMDC that help instruct and inform our practices for treating patients with metastatic renal cell carcinoma. The role of prognostic factors, cytoreductive nephrectomy, and outcomes from first/second/third-line therapies will be discussed. Now with new treatments and new mechanisms of action, new questions arise that large databases such as these and the Polish databases are well-suited to answer.

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MtDNA mutations and prostate cancer bone metastases Mutacje mitochondrialnego DNA a przerzuty do kości raka prostaty John A. Petros, Rebecca S. Arnold Emory University School of Medicine, Atlanta GA

We sequenced the mitochondrial genome in 10 prostate cancer patients with bone metastases enrolled in a rapid autopsy program. We then examined the number and location of somatically acquired mitochondrial DNA (mtDNA) mutations in the primary and two metastatic sites in each individual patient. Somatic mutations were significantly more numerous in bone compared to either the primary tumor or soft tissue metastases. A missense mutation at nucleotide position (np) 10398 (A10398G; Thr114Ala) in the respiratory complex I gene ND3 was the most common (7 of 10 patients) and was detected only in bone. Other notable somatic mutations that occurred in more than one patient include a tRNA Arg mutation at np 10436 and a tRNA Thr mutation at np 15928. The tRNA Arg mutation was restricted to bone metastases and occurred in three of 10 patients (30%). The genome was not mutated randomly. This indicates a pervasive selective pressure for bone metastatic cells that had acquired the 10398 mtDNA mutation. Two additional recurrent mutations (tRNA Arg and tRNA Thr) support the concept of bone site-specific “survival of the fittest” as revealed by variation in the mitochondrial genome and selective pressure exerted by the metastatic site.

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Role of oncogenic autophagy in clear cell renal cell carcinoma Rola autofagii w raku jasnokomórkowym nerki Maria F. Czyżyk-Krzeska1, Jarek Meller, David R. Plas 1

Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, 45267, USA

Metastatic ccRCC is largely incurable disease and the existing treatments targeting angiogenesis, tyrosine kinase receptors, and the mTOR pathway are only partially effective. We discovered a network in the regulation of autophagy and ccRCC oncogenesis that can be subject to experimental therapeutics. We found that loss of VHL promotes, in an HIF-independent manner, access to nutrients from intracellular sources through activation of LC3B-mediated autophagy. This autophagic program is necessary for tumor growth. This pathway, activated by the loss of VHL and miR204 tumor suppressors, involves oncogenic activity of the TRPM3 channel and its role in controlling autophagic programs through Ca 2+ and Zn2+ influx. A small molecule inhibitor of TRPM3, MFA, recapitulates the effects of genetic manipulations of TRPM3. This is an important proof-of-concept that TRPM3 represents an actionable target for treatment of ccRCC. Addiction of cancer cells to oncogenic autophagy creates a possibility to target other steps leading to formation of autophagosome by therapeutic interventions.

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Hypoxia alleviation: new treatments to normalize stably tumor vessels as they adjuvant therapeutic means Osłabienie hipoksji jako nowy sposób leczenia, normalizujący naczynia krwionośne, w terapii adiuwantowej Bouchra El Hafny-Rahbi1, Guillaume Collet1, Krzysztof Klimkiewicz1,2, Claudine Kieda1 Centre de Biophysique Moléculaire, UPR 4301 CNRS, Rue Charles Sadron, 45071 Orléans CEDEX2, France ; 2Jagiellonian University, Faculty of Biochemistry, Biophysics and Biotechnology, Ul. Gronostajowa 7, 30-387 Kraków, Poland 1

Oxygen partial pressure (pO2), is a key parameter of organ physiology, characterized by its own “normoxia” called: “physioxia”. It is disturbed in cancer “hypoxia” and turns on the angiogenic switch. O2 delivery remains impaired through the lack of blood flow and efficacy of the pathologic vessels. This compromises the chemotherapeutic treatments as well as radiotherapy. Revisiting the antiangiogenic strategies, counteracting tumor hypoxia to normalize vessels and allow them to be functional is an issue to control cancer microenvironment, avoid stem-like cells selection and cooperate to immunotherapy. Angiogenesis-related therapies favor blood vessels normalization/maturation vs destruction. Among them: tumor targeted genes expressed in a hypoxia-restricted manner to normalize tumor vasculature, restore efficient blood flow and reduce hypoxia. Endothelial precursor cells incorporating blood vessels normalize angiogenesis. Controlled tumor vessels normalization by enhancing O2 delivery using allosteric effectors of hemoglobin and control PI3K/AKT/mTOR pathway allowed stable tumor angiogenesis normalization. Data obtained from these strategies confirm that tumor hypoxia compensation helps radio- and/or chemotherapy, reverts pathologic angiogenesis, blocks metastasis and reverts tumor immune suppression.

Collet, G. et al. Cancer Ther. 13, 165-178, (2014). Kieda, C. et al. J. Mol. Med. 91, 883-899, (2013).

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What thyroid hormones have to do with renal cancer? Hormony tarczycy a rak nerki Agnieszka Piekiełko-Witkowska Centrum Medyczne Kształcenia Podyplomowego, Zakład Biochemii i Biologii Molekularnej. ul. Marymoncka 99/103, 01-813 Warszawa

Carcinogenesis is characterized by severe disturbances in key cellular processes such as proliferation, differentiation, apoptosis and metabolism. Remarkably, all these processes are regulated by thyroid hormones (TH) that act in virtually all human tissues. This implies that dysregulation of TH signalling may contribute to cancerspecific alterations of cellular processes. Intracellular TH concentrations are regulated by iodothyronine deiodinases (DIO1, DIO2, and DIO3), the enzymes that catalyse deiodination of iodothyronines. DIO1 and DIO2 contribute to the synthesis of 3,5,3' triiodothyronine (T3), while DIO3 reduces T3 concentration. The expression of DIO genes is disturbed in different cancer types. In clear cell renal cell carcinoma (ccRCC) the expression of DIO1 is dramatically reduced with concomitant decrease of intratumoural T3 concentration. Re-introduction of DIO1 into ccRCC cells in vitro results in inhibition of proliferation, along with reduced expression of genes controlling cell cycle, as well as inhibition of migration with concomitant changes in expression of adhesion genes, suggesting tumour-suppressive role of DIO1 in ccRCC. Opposite effects have been obtained for T3-inactivating enzyme, DIO3, in mice bearing xenografts of basal cell carcinoma and colon carcinoma. Thus, alterations in intracellular synthesis/degradation of TH actively contribute to the carcinogenic process, possibly offering new opportunities for cancer treatment.

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Saturday / Sobota 24.10.2015

Poster Session Sesja Posterowa

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MicroRNA-dependent regulation of expression of splicing factors in renal cell carcinoma (RCC) Zależna od mikroRNA regulacja ekspresji czynników splicingowych w raku nerkowokomórkowym (RCC) Joanna Bogusławska1, Elżbieta Sokół1, Beata Rybicka1, Katarzyna Rodzik1, Piotr Popławski1, Alicja Czubaty2, Hanna Kędzierska1, Zbigniew Tański3, Agnieszka Piekiełko-Witkowska1 1

The Centre of Postgraduate Medical Education, Department of Biochemistry and Molecular Biology, Warsaw, Poland, 2University of Warsaw, Institute of Biochemistry, Department of Biochemistry and Molecular Biology, Warsaw, Poland, 3Regional Hospital Ostrołęka, Poland

Renal cell carcinoma (RCC) is the most common type of renal cancer, characterized by high mortality and frequent metastasis. Splicing factors are key regulators of alternative splicing, actively contributing to carcinogenic process by influencing proliferation, apoptosis, adhesion, migration and invasion. Our previous studies revealed that expression of seven “classical” SR proteins: SRSF1, SRSF2, SRSF3, SRSF4, SRSF5, SRSF6 and SRSF7 as well as hnRNPA1 is disturbed in RCC. Here we hypothesized that disturbed expression of splicing factors in RCC may be caused by microRNAs: short, non-coding RNA molecules that regulate expression of genes at the posttranscriptional level via degradation of mRNA or inhibition of translation. Using bioinformatic predictions, molecular cloning methods, as well as functional assays (luciferase reporter assays, real-time PCR, ICC, WB) in RCC-derived cell line we found that indeed, the expression of splicing factors is regulated by microRNAs, namely: SRSF1 transcript is a target of miR-200c-3p, miR-214-3p, miR-10b-5p, miR-203 and miR-190a; SRSF2: miR-183-5p, miR-15a-5p, miR-16-5p and miR200c-3p; SRSF3: miR-135a-5p and miR-203; SRSF4: miR-34b-3p, miR-362-3p and miR-335; SRSF5: miR-181a; SRSF6: miR-192-5p, miR-214-3p and miR-200c-3p; SRSF7: miR-203, miR-216b and miR-30a-5p, whereas miR-149-5p, miR-16-5p, miR-135a-5p and miR-1 bind to 3’UTR of hnRNPA1. This regulation is mediated by miR-induced inhibition of translation (SRSF1 by miR-200c-3p and miR-214-3p; SRSF2 by miR-15a-5p and miR-16-5p; SRSF5 by miR-181a; SRSF7 by miR-203 and miR-216b; and hnRNPA1 by miR-16-5p) or degradation of target transcripts (SRSF1 by miR-10b-5p and miR-203; SRSF2 by miR-183-5p; SRSF3 by miR-135a5p and miR-203; SRSF4 by ; miR-34b-3p, miR-362-3p and miR-335-5p; SRSF6 by miR-192-5p, miR-214-3p and miR-200c-3p; SRSF7 by miR-30a-5p; and hnRNPA1 by miR-149-5p, miR-135a-5p, and miR-1). The expression of all these microRNAs was disturbed in RCC (n=33) tumour samples when compared witch paired-matched controls. To our knowledge this is the first study comprehensively describing the role

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of miRNAs in the regulation of splicing factors in RCC. The study was approved by Bioethical Committee of Centre of Postgraduate Medical Education. The project was supported by the programme of Polish Science Foundation: PARENT/BRIDGE cofinanced from European Union funds under Innovative Economy Operational Programme 2007-2013, Centre of Postgraduate Medical Education Grant (506-1-25-0114) and National Science Centre (2012/05/B/NZ5/01541).

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Screening for sphere-promoting culture conditions for RCC Poszukiwanie warunków do hodowli in vitro sfer RCC Klaudia K. Brodaczewska, Kamila Maliszewska-Olejniczak, Cezary Szczylik, Anna M. Czarnecka Department of Oncology with Laboratory of Molecular Oncology, Military Institute of Medicine, Warsaw

Introduction: Cancer stem cells (CSCs) are a population of rare cells that can mediate metastasis. They are an important target for treatment but are difficult to isolate and study in vitro. Aim of the study was to search for culture conditions that will allow to culture CSCs in renal cell cancer (RCC) basing on their ability to form spheres. Materials and methods: Established RCC cell lines: 786-O (primary), ACHN and Caki1 (metastatic) were cultured in standard tissue culture vessels in media that promote stem properties. PSC-400 and HEK293 lines were used as negative and positive controls, respectively. Cells were inspected each second day, for 2 weeks under inverted microscope to follow morphological changes and formation of pseudo-3D structures. Results: 7 out of 15 tested media showed sphere-promoting potential for RCC. Media that promoted creation of pseudo-3D structures were predesigned for culture of mesenchymal stem cells or human pluripotent stem cells with reduced level of serum or serum-free. Conclusions: Sphere-like structures can be induced in RCC cell lines in vitro with the use of commercially available media for culture of mesenchymal stem cells. Acknowledgements: The research was funded by a grant of NCBiR no LIDER/031/625/L-4/12/NCBR/2013. Corresponding author: [email protected]

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Evaluation of RCC subtypes in established cell lines Określanie podtypu raka nerki w liniach nowotworowych Klaudia K. Brodaczewska1, Mohammed I. Khan1, Aleksandra Fruba1, Irena Koch2, Cezary Szczylik1, Anna M. Czarnecka1 1

Department of Oncology with Laboratory of Molecular Oncology,, Military Institute of Medicine, Warsaw; 2Department of Pathomorphology, Institute of Mother and Child, Warsaw

Introduction: Molecular characterisation of renal cell cancer (RCC) allowed to unravel new pathways connected with disease pathology. This led to further subdivision of RCC and now renal neoplasms comprise a broad spectrum of histopathological entities described in 2004 WHO classification and modified by ISUP Vancouver Classification. Aim of the study was to categorise ccRCC (clear cell) cell lines obtained before 2004 into correct subtype as pRCC (papillary) was not distinguished at the time of cell line establishment. Materials and methods: RCC cell lines obtained since 1974 and recognised as ccRCC were used in the study. Expression of surface receptors characteristic for ccRCC and pRCC: CK10 and CK7 was performed by ICC. Mutation analysis of genes characteristic for ccRCC and pRCC: vhl and c-met was performed with genomic DNA. Results: ACHN, Caki1 and Caki2 cell lines expressed features of pRCC while 769-P and SMKT-R3 cells could not be categorised into any subtype with employed methods. Other cell lines were confirmed to be of clear cell histology. Conclusions: Exact subtype of RCC cell lines needs to be verified with modern molecular methods to fit novel disease classification. Acknowledgements: The research was funded by grants of NCN no UMO2014/13/B/NZ1/04010, NCBiR no LIDER/031/625/L-4/12/NCBR/2013 and WIM intramural no 1/8860 (347). Corresponding author: [email protected]

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Comparison of CTC isolation techniques in high-risk prostate cancer patients Porównanie technik izolacji CTC u pacjentów wysokiego ryzyka z rakiem stercza Joanna Budna1, Monika Świerczewska1, Agnieszka Jankowiak1, Wojciech Cieślikowski2, Maciej Zabel1 1

Department of Histology and Embryology, Poznan University of Medical Sciences; 2Department of Urology, Poznan University of Medical Sciences

The therapy of patients with high-risk PCA is a great challenge as over 50% of the surgically treated high-risk patients undergo a biochemical disease recurrence. Characterization of CTCs is promising method to understand biology of residual or relapsing PCA after local therapy. A number of new innovative technologies to improve methods for CTC detection with high sensitivity have recently been developed. Among them, the in vivo detector CANCER01 (Gilupi GmbH) and EPISPOT are of great attention. The aim of the study was to compare CTCs detection rates in both techniques.40 high-risk patients undergoing radiation were enrolled in the study. Detector coated with anti-EpCAM antibody was placed into the patient’s arm vein for 30 mins and selected EpCAM-positive cells were immunostained for presence of PSA, cytokeratins, nucleus and absence of leukocyte marker CD45. Presence of viable CTC was determined by EPISPOT based on PSA-secretion as well as FGF2 to demonstrate that a subset of CTCs also secrete this known stem cell growth factor relevant for the in vitro growth of micrometastatic cells. As we found at least 1 CTC in 50% of patients using detector CANCER01 and in 51% of patients using EPISPOT we conclude that both techniques have a high value and when applied together increase rate of viable CTC detection as prognostic marker in patients with minimal residual disease. Acknowledgements: The research was funded by a grant “ERA-NETTRANSCAN/03/2013”

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Możliwości pakietu Molecular Biology Data Analysis Toolkit (MBDat) udostępnianego w PCSS w ramach projektu PLGRID NG w badaniach biomedycznych Zuzanna K. Filutowska1,2,3, Katarzyna Grądzka1, Katarzyna Jasiewicz1 Uniwersytet Adama Mickiewicza, Wydział Biologii, Wydziałowa Pracownia Technik Biologii Molekularnej, 2Uniwersytet Adama Mickiewicza, Wydział Biologii, Instytut Biologii Molekularnej i Biotechnologii; 3 Instytut Chemii Bioorganicznej PAN, Poznańskie Centrum Superkomputerowo Sieciowe. 1

MBDat jest pakietem aplikacji, służącym do bioinformatycznych analiz biomedycznych, realizowanym w Poznańskim Centrum Superkomputerowo Sieciowym oraz na Wydziale Biologii Uniwersytetu im. Adama Mickiewicza w Poznaniu w ramach projektu PLGrid NG. Pakiet ten ma ona na celu udostępnienie infrastruktury obliczeniowej (gridowo-klastrowej) do analiz bioinformatycznych mających zastosowanie w naukach biomedycznych, zarówno w dziedzinie genomiki jak i proteomiki. białkami, zwłaszcza dla danych z eksperymentów wysokoprzepustowych (NGS). W skład pakietu wchodzą takie narzędzia jak Galaxy, Vector NTI, Chipster i Kepler.

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Analiza porównawcza i profilowanie sekwencyjne pozachromosomowego kolistego DNA (eccDNA) w jądrach ludzkich komórek nowotworowych i ich prawidłowych odpowiedników celem oznaczenia zmienności liczby kopii genów kodujących mikro RNA istotne w procesach nowotworzenia. Zuzanna K. Filutowska1,2,3, Katarzyna Grądzka1, Katarzyna Jasiewicz1 Uniwersytet Adama Mickiewicza, Wydział Biologii, Wydziałowa Pracownia Technik Biologii Molekularnej; 2Uniwersytet Adama Mickiewicza, Wydział Biologii, Instytut Biologii Molekularnej i Biotechnologii; 3 Instytut Chemii Bioorganicznej PAN, Poznańskie Centrum Superkomputerowo Sieciowe. 1

Projekt jest kontynuacją prowadzonych obecnie w Pracowni Technik Biologii Molekularnej UAM badań dotyczących detekcji i znaczenia jądrowego, pozachromosomowego DNA (eccDNA). Formy te są jednym z najważniejszych czynników odpowiedzialnych za zmienność liczby kopii genów (CNV) w komórkach prawidłowych jak również odgrywają one również istotną rolę w procesach nowotworzenia i starzenia się komórek. W wyniku prowadzonych badań wstępnych opracowano metodę izolacji i detekcji kolistych form DNA z tkanek ludzkich opartą na nowoczesnych technikach biologii molekularnej takich jak trawienie egzonukleolityczne, namnażanie za pomocą amplifikacji toczącego się koła (RCA) czy ilościowy PCR (qPCR). Celem niniejszego projektu jest analiza profili eccDNA wybranych linii komórek nowotworów złośliwych, stanowiących jedną z najczęstszych przyczyn zgonu kobiet w Polsce, oraz ich prawidłowych odpowiedników. Projekt zakłada izolację jądrowego eccDNA, z wybranych linii komórek rakowych oraz prawidłowych, pochodzących z szyjki macicy i jajników, za pomocą metody opracowanej wcześniej przez autorów projektu oraz poznanie ich sekwencji przy pomocy sekwencjonowania wysokoprzepustowego. Analiza bioinformatyczna uzyskanych sekwencji pozwoli określić jaki potencjał kodujący mają te formy DNA. Szczególny nacisk zostanie położony na wykrycie w sekwencjach eccDNA tych, które kodują mikroRNA istotne dla proliferacji, wzrostu i inwazyjności badanych typów nowotworów. Ostatecznym rezultatem projektu będzie weryfikacja hipotezy o wpływie zmienności genetycznej generowanej w postaci eccDNA na liczbę kopii genów odpowiedzialnych za kodowanie mikroRNA oraz utworzenie profili eccDNA w badanych liniach komórkowych.

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Inhibition of human renal cell carcinoma cell lines growth by cannabinoid receptor agonists Inhibicja ludzkich linii komórkowych raka nerkowokomórkowego przez agonistów receptorów kanabinoidowych. Mohammed I. Khan1*, Anna A. Sobocińska1,2, Igor Helbrecht1, Magdalena Król3, Cezary Szczylik1, Anna M.Czarnecka1 1

Department of Oncology with Laboratory of Molecular Oncology, Military Institute of Medicine, Warsaw, Poland; 2Faculty of Biology, Warsaw University, Warsaw, Poland; 3Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.

Introduction: Cannabinoids are the active components of Cannabis sativa and their derivatives. Cannabinoids involvement have been reported in inhibition of tumor growth in laboratory animals by inducing apoptosis of tumor cells and inhibiting tumor angiogenesis. Many of their anti-tumor effects are mediated through the cannabinoid receptors (CB1 and CB2) which comprises the endocannabinoid system (ECS). In this study, we evaluated the expression analysis of cannabinoid receptors using mRNA and protein level in renal cell carcinoma (RCC) cell lines. Selective cannabinoid receptor agonists was used for inducing cell apoptosis and cell proliferation was analysed using alamar blue cell viability assay. Materials and methods: Primary and metastatic RCC cell lines were used. Selective CB1 and CB2 agonists were used for inducing cell apoptosis. Cell proliferation was analysed using alamar blue cell viability assay. Results: Our preliminary observation shown that CB1 receptor is down-regulated in metastatic RCC cell lines in comparison to CB2 receptor. Furthermore, in primary RCC cell lines both CB1 and CB2 receptors expression were observed. Conclusion: Cannabinoid receptors agonist are highly selective for exerting antiproliferative effects on RCC cell lines. Acknowledgments: This work was funded by the National Science Centre (NCN) grant “PRELUDIUM”, grant no. UMO-2013/09/N/NZ5/02809. *Corresponding author: [email protected]

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Gene expression profiling of mesenchymal stem cells CD105+ as CSCs/TICs isolated from primary and metastatic renal cell carcinoma Profilowanie ekspresji genów z mezenchymalnych komórek macierzystych CD105+ jako CSCs/ TICs wyizolowane z pierwotnego i przerzutów raka nerki Mohammed I. Khan1*, Anna M. Czarnecka1, Sławomir Lewicki2, Igor Helbrecht1, Klaudia Brodaczewska1,4, Robert Zdanowski2, Magdalena Król3, Cezary Szczylik1 1

Department of Oncology with Laboratory of Molecular Oncology, Military Institute of Medicine, Warsaw, Poland; 2Department of Regenerative Medicine, Military Institute of Hygiene and Epidemiology, Warsaw, Poland; 3Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences - WULS, Warsaw, Poland; 4Department of Parasitology, University of Warsaw, Warsaw, Poland.

Introduction: Recent advancement in cancer research has shown those cancers are composed of heterogeneous population of cancer cells and there is small population called "cancer stem cells/ tumor initiating cells (CSCs/TICs)" hiding in the vicinity of such tumor mass. To get insight into renal cell carcinoma (RCC) pathogenesis and possibly propose novel targeted therapies using CSCs/TICs gene expression profiling, isolation and effective propagation of RCC-cancer stem cells is essential. This research was designed to optimize cell culture techniques in the field of renal cancer and characterize biology and gene expression of renal cancer stem cells (RCC-CSC/TICs). Materials and methods: CD105+ cells from primary (CAKI-2) and metastatic (ACHN) RCC cell lines were isolated using FACS. Gene expression profiling was performed on isolated CD105+ cells using Agilent microarrays. Ingenuity pathways analysis (IPA), Gene Ontology (GO) and KEGG pathway enrichment was performed on differentially expressed genes from CD105+ cells. Results: 1411 genes were in common differentially (up-/down-regulated) expressed between CD105+ cells from primary and metastatic RCC. RCC-CSCs consist of different altered signaling pathways. The common altered pathways in CD105+ cells are TGF-β, Wnt/β-catenine and Epithelial-Mesenchymal Transition (EMT). TGFB1, ERBB2, and TNF are most significant transcriptional regulators in RCC-cancer stem cells. Conclusion: These data may be useful for further signalling pathway studies and provide insight into the molecular mechanism of RCC using CSCs/TICs as a platforms for pre-clinical drug application analysis. Acknowledgments: This work was funded by the Foundation for Polish Science TEAM project TEAM/2010-6/8. *Corresponding author: [email protected]

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Developing experimental model approach for overcoming resistance of renal cell carcinoma cells to mTOR and tyrosine kinase inhibitors

Anna Kornakiewicz1,2, Anna M. Czarnecka1, Cezary A. Szczylik1 1

Military Institute of Medicine, Warsaw, Poland; 2Postgraduate School of Molecular Medicine, Warsaw, Poland

Introduction: To break resistance of renal cancer cells with mTOR inhibitors simple experimental model for selecting resistant cancer cells is necessary for assessment of dynamic changes in mTOR network components expression. Materials and Methods: Primary tumor and metastatic cell lines and stem-like cells line were analyzed with Alamar Blue Assay at the beginning of experiment and changes in IC50 were continuously assessed in the course of everolimus treatment with the dose near IC50. Different computational and mathematical models were analyzed to redefine experimental model. Results: Points of time at which receptors expression will be measured in further experiments were chosen. Higher IC50 was characteristic not only for metastatic lines, but also for cRCC lines with CD133+ marker. Similar tendency in case of IC50 values in different cell lines was observed in the course of everolimus treatment. Here, we show one of approaches to select resistant cells with the dose of drug close to IC50 and compare effect of mTOR inhibitor: everolimus with TKIs: suntinib, axitinib and sofrafenib on primary tumor and stem-like renal carcinoma cells. Conclusions: We indicate possible usefulness of the model of tumor progression based on complex automata driven by particle dynamics to redefine experimental model. In turn, the experimental model may improve computational model of tumor growth and angiogenesis. Acknowledgements: The research was funded within the program under the name "Diamond Grant" no. DI2012 007842 and supported by the Foundation’s for Polish Science TEAM project no. TEAM/2010-6/8.

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Planowanie brachyterapii raka prostaty z wykorzystaniem Tomografii Komputerowej u chorych po pierwotnej całkowitej amputacji odbytu Łukasz Kowalik1, Jarosław Łyczek1, Damian Kazalski1, Marcin Sawicki1, Mirosław Kolbusz2 Zakład Brachyterapii, Szpital Specjalistyczny w Brzozowie, Podkarpacki Ośrodek Onkologiczny im. Ks. B. Markiewicza; 2Zakład Radioterapii, Szpital Specjalistyczny w Brzozowie, Podkarpacki Ośrodek Onkologiczny im. Ks. B. Markiewicza 1

Materiały: W grupie 9 chorych pierwotnie leczonych radykalnie z powodu raka odbytu z zastosowaniem teleradioterapii wysokodawkowej oraz radykalnej operacji sposobem Milesa, u których rozpoznano miejscowo zaawansowanego raka prostaty przeprowadzono brachyterapię HDR. Metody: U chorych zastosowano frakcjonowaną brachyterapię HDR podając w trakcie leczenia 3 x 12Gy w odstępie 21 dni. Do lokalizacji gruczołu krokowego, aplikatorów oraz do planowania leczenia wykorzystano tomograf komputerowy (CT). Ze względu na ruchomość gruczołu krokowego w zależności od ułożenia pacjenta, wybrano pozycję na boku zmieniając położenie przy każdej frakcji. Aplikacji dokonywano w znieczuleniu podpajęczynówkowym. Wstępna tomografia pozwalała na lokalizację gruczołu krokowego, pęcherza moczowego oraz struktur kostnych. Powtarzana po założeniu kolejnych 3 igieł pozwalała na określenie ich położenia oraz zaplanowanie umiejscowienia i kolejnych igieł. Po pełnej implementacji badanie CT wykorzystane zostało do planowania leczenia w systemie Oncentra Brachy 4.1. Wyniki: Zastosowanie techniki wykorzystania Tomografii Komputerowej w planowaniu leczenia brachyterapii HDR nowotworu gruczołu krokowego pozwoliło na objęcie narządu izodozą V100% w zakresie 87 – 98% (mediana 91%). W dwóch pierwszych przypadkach nie uwidoczniono dostatecznie dobrze przebiegu cewki moczowej, w pozostałych po zastosowaniu kontrastów jodowych w cewniku Foleya można było zdefiniować dokładny jej przebieg. We wszystkich wypadkach pęcherz moczowy był dostatecznie widoczny. Wnioski: Brachyterapia HDR z wykorzystaniem tomografii komputerowej oraz systemu planowania leczenia Oncentra Brachy 4.1 pozwala na leczenie pacjentów z nowotworem prostaty po usunięciu odbytnicy. Optymalizacja planu leczenia pozwala na zachowanie zaleceń ICRU. Wykorzystanie tomografii komputerowej do planowania brachyterapii HDR raka prostaty powinno być techniką zalecaną u chorych po amputacji brzuszno-kroczowej odbytu.

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Phenotypic and molecular changes under reciprocal interaction of normal and renal cancer cells Fenotypowe i molekularne zmiany pod wzajemnym wpływem komórek zdrowych i nowotworowych nerki Paweł Krasowski1,2, Katarzyna Kaminska1, Anna M. Czarnecka1, Michał Marcinkowski2, Elżbieta Grzesiuk2, Cezary Szczylik1 1

Military Institute of Medicine, Oncology Clinic, Warsaw, Poland; 2Institute of Biochemistry and Biophysics PAS, Department of Molecular Biology, Warsaw, Poland

Introduction: Interleukin-6 (IL-6) - pleiotropic cytokine with potential antitumor activity, was suggested to have a stimulatory growth effect in renal cell cancer (RCC) tumors. Obtaining primary data suggesting that IL-6 is produced at high levels by renal cell carcinoma cell lines. We aim to investigate the molecular mechanisms involved in its possible role as an autocrine growth factor. We hypothesized that in metastatic clear cell RCC the complex of interleukin-6 and its soluble receptor (IL6sR; complex IL- 6/IL-6sR) play a key role in this process using signal transduction pathway of gp130/STAT3. The aim of the study was identification of the contribution IL-6/IL- 6sR complex and its signal transduction pathway in the communication of renal cell carcinoma and cells of the target tissue in metastases. Materials and methods: Cell lines of healthy lungs (NL-20) were cultured with conditioned medium obtained from renal cancer cell lines (Caki-1, Caki-2, ACHN). Results: Results shown that IL-6 secreted from cancer cell lines and proteins of its signaling pathway influence the biology of healthy cells. In turn IL-6 creates formation of microenvironment favoring tumor niche. Nevertheless alternative signal transduction pathway other than gp130/STAT3 is responsible for this phenomenon. Acknowledgements: This research has been supported with the National Science Centre project no. UMO-2011/01/B/NZ4/01602. Special thanks for FODiK for buying necessary reagents.

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MCPIP1 and HIF2α interplay and regulation of angiogenesis in clear-cell renal cell carcinomas Współdziałanie MCPIP1 i HIF2α w regulacji angiogenezy w jasnokomórkowym raku nerki Janusz Ligęza*1, Natalia Gach*1, Barbara Lipert1, Katarzyna Miekus1, Wacław Wilk2, Janusz Jaszczynski2, Andrzej Stelmach2, Agnieszka Loboda3,4, Józef Dulak3,4, Wojciech Branicki1, Janusz Rys2, Jolanta Jura1 * This authors equally contributed to this work 1

Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland; 2Centre of Oncology, Maria Skłodowska-Curie Memorial Institute, Krakow, Poland; 3Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland; 4Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland

Hypoxia, a characteristic feature of locally advanced solid tumors is important factor in cancer progression and resistance to therapy. The main family of transcription factors responsible for cell survival and adaptation in hypoxia are the “hypoxiainducible factors” (HIFs). HIFs regulates tumor angiogenesis, proliferation, invasion, metastasis and additionally are involved in inflammation, the “seventh” hallmark of cancer. In our study we analyzed the effect of oxygen supply on expression of selected genes involved in regulation of inflammatory pathways in Clear Cell Renal Cell Carcinoma cell line, Caki-1. One of the genes regulated in low oxygen conditions was ZC3H12A encoding MCPIP1 protein which has RNase properties and regulates the half-life of transcripts coding for pro-inflammatory cytokines. We found that MCPIP1 is downregulated in hypoxia and that HIF2α is responsible for this regulation. Moreover, MCPIP1 negatively regulates HIF2α what creates negative loop between these two molecules. Overexpression of MCPIP1 in Caki-1 cells leads to strong downregulation of transcripts encoding VEGFA, IL-6 and GLUT1. Furthermore, MCPIP1 influences activity of key signaling pathways. These observations suggest that MCPIP1 is involved in regulation of angiogenesis in ccRCC. Acknowledgements: This study was supported by National Science Center grant: 2011/03/B/NZ1/00023. Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University is a partner of the Leading National Research Center (KNOW) supported by the Ministry of Science and Higher Education.

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Obustronny rak nerek. Dwuetapowa resekcja guzów Lech Lipiec, Przemysław Dudek, Jacek Adamczyk, Michał Lipowski Oddział Urologii Szpital Specjalistyczny im. św. Łukasza w Końskich

Wprowadzenie: Nefrektomia radykalna jest nadal złotym standardem postępowania w przypadku nowotworów o średnicy powyżej 4 cm. Operacje nerkoszczędzajace są wskazane jako metoda leczenia w przypadku nerki jedynej lub obustronnych guzów nerek Doniesienia potwierdzają jednak możliwość skutecznego leczenia NSS w przypadku nowotworów o średnicy nawet powyżej 4 cm. Cel pracy: Przedstawienie sposobu postępowania w przypadku chorej z obustronnymi dużymi guzami nerek. Materiały i metody: 75-letnia chora została przyjęta do Oddziału Urologii w maju 2007 r. z powodu złego samopoczucia oraz stopniowej utarty masy ciała. W badaniach obrazowych /USG, CT/ stwierdzono obustronne guzy nerek. W nerce lewej guz o wymiarach 7,5 x 6 x 5,5 cm zlokalizowany w górnym biegunie, ale w pobliżu wnęki nerki. W nerce prawej guz o wym. 6 x 5,5 x 5 cm położony w górnym biegunie oraz dodatkowo guzek nadnercza prawego o śr. 1 cm. Po konsultacji nefrologicznej i endokrynologicznej i wykonaniu badania naczyniowego, zadecydowano o obustronnej dwuetapowej operacji NSS z usunięciem nadnercza po stronie prawej. Pierwszy usunięto guz nerki lewej (zimne niedokrwienie ok. 32 min). Ze względu na niekorzystne położenie resekowano z guzem fragment żyły nerkowej. Drugą operację przeprowadzono po miesiącu – resekowano górny biegun nerki prawej wraz z nadnerczem (niedokrwienie zimne ok. 22 min). Wyniki: Przebieg operacji i pooperacyjny bez powikłań. Po pierwszej operacji wystąpiła przemijająca niewydolność nerek z maksymalnymi wartościami stężeń mocznik: 65 mg/ dl, kreatynina 1,8 mg/dl. Wynik histopat. Ca. calarocellulare NG 4. Po drugiej operacji obserwowano podwyższenie stężenia mocznika do 104,1 mg/dl oraz kreatyniny do 3,24. Przy wypisie mocznik 65 mg/dl, kreatynina 2,98 mg/dl. Wynik histopat. Ca. clarocellulare NG 2. W nadnerczu–incydentaloma. W trakcie 8 letniej obserwacji (USG, CT, RTG klp., LDH) nie stwierdzono cech wznowy miejscowej ani przerzutów odległych. Aktualne stężenie kreatyniny wynosi 2,3 mg/dl. Chora żyje bez dializ w dobrym stanie ogólnym. Wnioski: W wybranych przypadkach można wykonać bezpiecznie resekcję obustronnych guzów nerek o śr. powyżej 4 cm z dobrym wynikiem onkologicznym i czynnościowym nerek. Metodę nerkooszczędzającą należy rozważyć w przypadku obustronnych guzów nerek. Konieczna jest wnikliwa diagnostyka także z badaniem naczyniowym oraz ścisła współpraca z oddziałem nefrologicznym.

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Brachyterapia HDR jako intrygująca propozycja w leczeniu wznowy miejscowego raka prostaty Jarosław Łyczek, Damian Kazalski, Łukasz Kowalik, Iwona Kubicka-Mendak Centrum Onkologii – Instytut im. Marii Curie-Skłodowskiej Warszawa, Szpital Specjalistyczny Podkarpacki Ośrodek Onkologiczny Brzozów, Świętokrzyskie Centrum Onkologii Kielce

W okresie 2000-2013 leczono łącznie 213 chorych z rozpoznaniem wznowy miejscowej raka gruczołu krokowego, po uprzednim leczeniu radykalnym. Ze względu na różnorodność technik stosowanych w leczeniu tej grupy chorych, z analizy wyłączono wszystkich chorych leczonych innym schematem niż 3x10Gy co 21 dni. Do analizy włączono grupę 168 chorych, u których stwierdzono we wstępnej kwalifikacji do leczenia pierwotnego stopień Gleasona 6 u 60% chorych, stopień Gleasona 7 u 22% chorych, stopień zaawansowania T2 u 68% chorych, T1 u 27%. Pierwotne leczenie u 71% chorych opierało się na radykale radioterapii, a u 23% na chirurgii skojarzonej z radioterapią. W analizowanej grupie zdecydowana większość wznów obserwowana była między 22 a 27 miesiącem leczenia. Średni okres obserwacji w analizowanej grupie wyniósł 72 miesiące. 5-letnie przeżycia całkowite wyniosły 67%, a 5-letnie przeżycia wolne od choroby 58%. Analizując poszczególne czynniki, tj. stopień zaawansowania, wyjściowy poziom PSA, czas do wznowy, czy stopień w skali Gleasona stwierdzono silną zależność pomiędzy poziomem odpowiedzi a skalą Gleasona i istotnie statystyczną zależność od czasu, który upłynął od leczenia pierwotnego do wznowy. Zarówno stopień zaawansowania miejscowego, jak i poziom wyjściowego PSA nie wykazały istotności statystycznej. Brachyterapia HDR wydaje się być efektywnym i nie obciążonym powikłaniami, zarówno wczesnymi jak i późnymi, sposobem postępowania u chorych z miejscową wznową raka prostaty po leczeniu radykalnym.

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Limitation of collagenase 1 and 3 content in ccRCC of human kidney Ograniczenie zawartości kolagenazy 1 i 3 w raku jasnokomórkowym ludzkiej nerki Grzegorz Młynarczyk1,2, Lech Romanowicz1, Barbara Darewicz2, Krzysztof Sobolewski1 1

Department of Medical Biochemistry, Medical University of Bialystok; 2 Department of Urology, Medical University of Bialystok

Matrix Metalloproteinases are a group of endopeptidases that contain a zinc and calcium atoms in molecule. They are divided into collagenases, gelatinases, stromelysines, matrylisynes, membrane-type metalloproteinases and some specific enzymes like elastase. Their main function is primary degradation of extracellular matrix components. In carcinogenetic processes metalloproteinases are involved in local invasion of tumor and metastasis. Clear-cell kidney cancer is the most frequent type of renal cell carcinoma. It originates in the lining of the proximal convoluted tubes and gives most frequent metastasis to lungs and bones. The aim of the study was estimation of content and activity of matrix metalloproteinases in human kidney not-changed and changed by carcinogenetic process. The study group comprised 20 clear-cell kidney cancer patients at the G2 and G3 Fuhrman scale who underwent radical surgical treatment. The study material included cutaway of primary tumors and healthy tissue collected during the surgical procedure. The content of metalloproteinases,MMP-1 (collagenase 1), MMP-13 (collagenase 3) and MMP-2 (gelatinase A), was determined with the immunoenzyme ELISA tests. Gelatinolytic activity was detected in sample tissue homogenates using zymography. Significantly lower content of all investigated metalloproteinases in tumor tissues than in healthy tissues was observed. Both control tissues and clear-cell kidney cancer at G2 showed similar activity of gelatinase A. Higher MMP-2 activity in tumor at G3 tissue was found. Much lower content of matrix metalloproteinases in malignant tumors, especially both investigated collagenases, in comparison to healthy part of renal tissue suggests that clear-cell kidney cancer may block an expression of investigated enzymes and thereby may differ from other malignant tumors.

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The Influence of Insulin and IGFs on Renal Cancer Cells Wpływ insuliny i insulino-podobnych czynników wzrostu na komórki raka nerki Wojciech Solarek1,2, Cezary Szczylik1, Anna M. Czarnecka1 Department of Oncology, Military Institute of Medicine, Szaserów 128, 04-141 Warsaw, Poland , 2 School of Molecular Medicine, Medical University of Warsaw, Warsaw, Poland 1

Introduction: Renal cell carcinoma (RCC) is the 10th malignancy worldwide and the most frequent type of kidney cancer in adults. Insulin-like growth factor receptor (IGF-IR), a tyrosine kinase receptor for insulin-like growth factors IGF-I and IGF-II, has been well documented in cell culture models, animal studies, and humans to play a role in malignant transformation, progression and metastasis. Materials and methods: In this study we showed renal cancer cells-insulin molecular patophysiology in metastatic RCC in vitro model. It was verified how insulin or insulin-like growth factors impact RCC cells proliferation, migration or metastatic potential by Alamar Blue assay and Wound Healing Assay. Results: Both IGFs and insulin can affect RCC cells growth and migration potential. IGF-1R inhibition leads to decrease in most of RCC cell lines growth but do not inhibit growth of HEK293 cell line probably through the IR dependent mechanism. RCC cell lines express IGF-1R and are responsive to IGFs stimulation but do not express IR thus insulin in high concentrations may influence their growth through IGF-1R receptor stimulation. Conclusions: IGF-1 and IGF-2 through IGF-1R can affect cell stimulation pathways, what may be one of the mechanism promoting renal cancer growth and progression Acknowledgements: This research was supported by National Science Centre UMO2012/05/D/NZ5/01844

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Notatki

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