SORAFENIB BLOCKS TUMOUR GROWTH, ANGIOGENESIS AND METASTATIC POTENTIAL IN PRECLINICAL MODELS OF OSTEOSARCOMA
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1 SORAFENIB BLOCKS TUMOUR GROWTH, ANGIOGENESIS AND METASTATIC POTENTIAL IN PRECLINICAL MODELS OF OSTEOSARCOMA Ymera Pignochino, PhD Division of Medical Oncology IRCC - Candiolo - TO
2 CLINICAL FEATURES OF OSTEOSARCOMA (OS) OS is the most common primary malignant bone tumour in children and young adults OS is characterized by an aggressive clinical course Since 1980s multiagent chemotherapy significantly increased 5-year survival of localized OS patients from 20 % to approximately 65% Pulmonary metastases, central presentation and local non-resectable relapse still cause a fatal outcome Both novel chemotherapeutic drugs and radiometabolic therapy based on samarium failed to improve overall survival The search for alternative agents in OS is therefore mandatory.
3 OSTEOSARCOMAS EXPRESS ALTERED LEVEL OF PATHWAYS ACTIVATIONS P-ERK / 30 (67%) MCL-1 24 / 30 (80%) P-ERM 21 / 30 (70%)
4 MOLECULAR TARGETS OF SORAFENIB Raf-1 BRAF wild type and V600E mutated Pro-angiogenic RTKs : VEGFR-2 e 3 PDGFR-β RTKs involved in tumor progression: FLT-3 c-kit FGFR-1 Ret Mcl-1 anti-apoptotic protein expressed in different cancer cell lines
5 SORAFENIB INHIBITS OSTEOSARCOMA CELL LINE PROLIFERATION IN A DOSE AND TIME DEPENDENT MANNER SJSA-1 viability test N viab le cells / co n tro l Dose-effect curve 0 0h 24h 48h 72h time point e f f e c t µM 7.5µM 5µM 2.5µM peg % inhibtion of Normal HOB at doses>10µm sorafenib Dose
6 SORAFENIB INHIBITS CELL CYCLE PROGRESSION AND INDUCES APOPTOSIS IN OS CELL LINES
7 SORAFENIB BLOCKS ERK PHOSPHORILATION AND MCL-1 EXPRESSION IN A DOSE DEPENDENT MANNER
8 SORAFENIB DOWNREGULATES EZRIN PHOSPHORILATION SOR (µm)
9 SORAFENIB DISPLAYED ANTITUMORAL ACTIVITY AGAINST OS XENOGRAFTS IN SCID MICE SOR 100 NT mean volume (mm 3 ) Sorafenib 100 mg/kg/die Sorafenib 30 mg/kg/die Sorafenib 10 mg/kg/die vehicle days after treatment
10 IN VIVO EFFECT OF SORAFENIB Sorafenib reduces: - tumor cellularity Hematoxylin and Eosin - blood vessel number Masson s trichromic staining
11 ANTIANGIOGENETIC EFFECT OF SORAFENIB IN CHORIOALLANTOIC MEMBRANES (CAM) OS cells conditioned supernatants induce angiogenesis in CAM + sorafenib Sorafenib blocks angiogensis induced by OS supernatants in CAM
12 SORAFENIB REDUCES INVASIVENESS AND NEOANGIOGENETIC POTENTIAL Elisa test on 7 OS cell lines
13 SORAFENIB REDUCES LUNG FOCI IN EXPERIMENTAL MODELS OF OS METASTASES NT SOR SJSA-1 cells injected into the tail vein of SCID mice give rise to tumor colonies in lungs after 3 weeks. Additional 16 days of sorafenib treatment strikingly reduces focal area in respect to untreated controls
14 CONCLUSIONS AND PERSPECTIVES: CLINICAL TRIAL Phase II study, open label, non-randomized study of second line treatment with sorafenib (BAY ) in Patients affected by relapsed high-grade osteosarcoma. Test Drug: Sponsor s Name and Address: Sorafenib Italian Sarcoma Group c/o Istituti Ortopedici Rizzoli Via di Barbiano, 1/ Bologna Study chairmen: M. Aglietta, G Grignani Sponsor s Telephone Number: Study HGosteo-BAY versione 1.0 Development Phase: Phase II
15 SORAFENIB DISPLAYED ANTI TUMORAL PARTICIPATING ACTIVITY AGAINST UNITS OS XENOGRAFTS IN SCID MICE Division of Medical Oncology- IRCC Candiolo Prof. M Aglietta, Dr G. Grignani Pediatric Onco-hematology -OIRM Sant anna Dr Franca Fagioli Division of Medical Oncology- Istituti Ortopedici Rizzoli Prof P. Picci, S Ferrari Istituto Tumori di Milano Prof P. Casali
16 STUDY OBJECTIVES Primary: To document the clinical activity of sorafenib as single agent in term of percentage of patients with advanced high-grade osteosarcoma free from progression at 4 months (PFS) Secondary: To evaluate the response rate, the clinical benefit, and overall survival of patients and the correlation between sorafenib-targets (Raf-1, VEGFR and PDGFR-β) expression and clinical outcome. To explore different than RECIST criteria to evaluate response such as Choi criteria
17 INCLUSIONS CRITERIA 1. Patients with histologically documented and not surgically resectable or metastatic highgrade osteosarcoma which progressed after first line or second line treatments. 2. Age > 15 yrs old 3. Other criteria were standard
18 ENROLLMENT AND ACCRUAL 35 PATIENTS planned Accrual as expected Last patients enrolled in December 2009 Toxicity as expected Analysis of the data for activity planned in June 2010
19 ACKNOWLEDGEMENTS Massimo Aglietta Giovanni Grignani Sandra Aliberti Manuela Motta Bruno Torchio Loretta Gammaitoni Giuliana Cavalloni Carmine dell Aglio Marco Basiricò Alessia Bottos Federico Bussolino Stefania Bruno Marta Tapparo Giovanni Camussi Piero Picci Stefano Ferrari Marco Alberghini Franca Fagioli
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