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The EurOPDX Consortium: Objectives, Achievements & Future Directions in developing PDXs and derivatives Emilie Vinolo 1* , Els Hermans 2 , Laura Soucek 3,4,5 , Denis G. Alférez 6 , Frederic Amant 1 , Daniela Annibali 1 , Joaquín Arribas 3,4,5 , Mohamed Bentires-Alj 7 , Cristina Bernadó 3,4,5 , Andrea Bertotti 8 , Andrew V. Biankin 9 , Alejandra Bruna 10 , Eva Budinsk 11 , Annette T. Byrne 12 , Carlos Caldas 10 , Oriol Casanovas 13 , David K. Chang 9 , Robert B. Clarke 6 , Simona Corso 8 , Georges Coukos 14 , Virginie Dangles-Marie 15 , Didier Decaudin 15 , Jeroen Depreeuw 1 , Zdenka Dudová 11 , Olivier Elemento 16 , Silvia Giordano 8 , Eva Gonzalez-Suarez 17 , Manuel Hidalgo 18 , Georgio Ga. Inghirami 16 , Monika Jarzabek 12 , Steven de Jong 19 , Jos Jonkers 20 , Kristel Kemper 20 , Aleš Křenek 11 , Martin Kuba 11 , Luisa Lanfrancone 21 , Pedro P. Lpez Casas 18 , Gunhild Mari Mælandsmo 22 , Elisabetta Marangoni 15 , Enzo Medico 8 , Ian Miller 12 , Kim Moran-Jones 9 , Beatriz Morancho 3,4,5 , Fariba Nemati 15 , Jens Henrik Norum 22 , Héctor G. Palmer 3 , Daniel S. Peeper 20 , Pier Giuseppe Pelicci 21 , Alejandro Piris-Giménez 3 , Miguel Angel Pujana 13 , Sergio Roman-Roman 15 , Oscar M. Rueda 10 , Joan Seoane 3,4,5 , Violeta Serra 3 , Alexandros Sigaras 16 , Sabine Tejpar 1 , Martin Tomas 11 , Livio Trusolino 8 , Ate van der Zee 19 , Marieke van de Ven 20 ; Dominique Vanhecke 14 , Alberto Villanueva 13 , Bea Wisman 19 1 seeding science, Paris, France; 2 University Hospital Leuven, Belgium; 3 Vall d'Hebron Institute of Oncology, Barcelona, Spain; 4 Universitat Autònoma de Barcelona, Bellaterra, Spain; 5 Institució Catalana de Recerca I Estudis Avançats (ICREA), Barcelona, Spain; 6 Manchester Cancer Research Center, University of Manchester, UK 7 University of Basel / University Hospital Basel, Switzerland; 8 Universitof Torino, Candiolo Cancer Institute IRCCS, Candiolo, Italy; 9 Wolfson Wohl Cancer Research Center, Institute of Cancer Sciences, University of Glasgow, UK; 10 Cancer Research UK Cambridge Institute, Cambridge Cancer Centre, UK; 11 Masarykova Univerzita, Brno, Czech Republic; 12 Royal College of Surgeons in Ireland, Dublin, Ireland; 13 Catalan Institute of Oncology, L’Hospitalet de Llobregat, Barcelona, Spain; 14 Lausanne Branch, Ludwig Institute for Cancer Research at the University of Lausanne, Switzerland; 15 Institut Curie, Paris, France; 16 Weill Cornell Medical College, New York, NY, USA; 17 Bellvitge Biomedical Research Institute IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain; 18 Centro Nacional de Investigaciones Oncolgicas, Madrid, Spain; 19 University Medical Center Groningen, University of Groningen, The Netherlands; 20 Netherlands Cancer Institute - Antoni van Leeuwenhoek, Amsterdam, The Netherlands; 21 Istituto Europeo di Oncologia, Milan, Italy; 22 Oslo University Hospital, Oslo, Norway. Started in early 2013, the EurOPDX Consortium now includes 18 academic institutions throughout Europe and in the US studying patient-derived tumours grown as xenografts in mice (PDX), to challenge the extreme inter- and intra-tumour heterogeneity of biological behaviour and treatment response in human cancer. EurOPDX involves world-renowned experts at the forefront of research in basic, preclinical, translational and clinical oncology across multiple pathologies, and displays a wide range of expertise in technological platforms. The distributed PDX collection of the consortium consists of more than 1,500 subcutaneous and orthotopic models for over 30 different pathologies (details of the collection on www.europdx.eu). Despite the great promise for PDX to improve the attrition rate in cancer, generalized use in high-throughput drug screenings is unrealistic, from an animal welfare point of view as well as for financial reasons. Short-term 2D and propagateable 3D (organoids) ex vivo cultures from PDX-derived tissue, retain genomic features and in vivo drug response and can systematically and consistently be generated from PDXs. This non-animal and cost effective drug development pipeline for high-throughput screening prior to in vivo testing using PDX models is currently still under investigation across cancer types and open for improvement. The primary goal of the consortium is to increase the visibility of this collection, and share the established PDX models and derivatives with the academic scientific community. In addition, our objective is to improve the predictability of preclinical data by: (i) improving characterisation of the models (ii) implementing new PDX models to strengthen the representativeness of the collection, and humanized PDX models (iii) harmonising working practice (iv) integrating complementary models (e.g. ex vivo 2D and 3D assays) in innovative drug development pipelines, and (v) leveraging deep molecular and pharmacogenomic profiling data of the collection of models to discover predictive biomarkers, new targets, and new strategies to overcome drug resistance. ABSTRACT / OBJECTIVES OF EurOPDX A NETWORK OF TRANSLATIONAL AND CLINICAL RESEARCHERS * Contact for further information: [email protected] STANDARDS for REFINEMENT • Establishing common standards increase animal welfare and improve reproducibility of preclinical results. • Workshop Nov. 2014: Common standards for the minimal validation and characterisation of PDX models. • Generalisation of PDX fingerprinting as quality control. • Ongoing Installation of the Laboratory Assistant Suite (LAS) developed at the University of Torino across several EurOPDX centers. LAS is a laboratory information management system for live tracking by digital barcoding of all biobanking information and in vivo data. ACHIEVEMENTS & ACTIVITIES TOWARDS THE 3Rs SHARING MODELS for REDUCTION • Increase visibility of established models, across the Consortium and towards the scientific community, to avoid duplication of efforts and make maximal used of available models. • Models currently accessible on a collaborative basis for the moment. • Ongoing Set-up of a common and public database on cBioPortal (release of pilot database foreseen for early 2017), which will include clinical information and molecular characterisation of the models. Data on PDX cohorts will be included progressively. Initiatives towards the “professionalization” of models sharing (biobanking/trans-national access to the collection). OTHER NETWORKING ACTIVITIES for REDUCTION • Many collaborations initiated since the start of EurOPDX, sharing of negative results, of protocols. • 1 st educational workshop in Weggis, Switzerland on October 3-6, 2016 to open up discussions with the European and international community, in particular young researchers. METHODOLOGY Engraftment Phase (F1) Expansion (F2) and validation (F3) phase www.europdx.eu/ ESTABLISHMENT OF PDXS Transplantation of a patient’s tumour sample in immunocompromised mice Following positive engraftment, expansion over several “passages” in mice BENEFITS OF PDX MODELS PDXs maintain the characteristics of the original patient’s tumour at the histological, genetic, and pharmacological levels Intra-tumour heterogeneity is largely maintained from the original patient’s tumour Collections of PDXs / “xenopatients” allow to represent the extreme inter- and intra-tumour heterogeneity of biological behaviour and treatment response in human cancer Personalised medicine approaches (biomarker analyses, Avatar and proxy approaches…) > Much improved predictability for success in the clinics as compared to cancer cell lines or cell lines xenografts, ex vivo PDX-derived 2D and 3D (organoid) cell cultures for high- throughput drug screening LIMITATIONS OF PDX MODELS Immunocompromised mice lack a functional immune system. > Development of “humanised” PDX models Mid- to High-throughput screening of single drugs or drug-drug combinations in collection of models is not ethical nor feasible. > Development of alternative study designs (1 animal per drug per model) and alternative ex vivo approaches DEVELOPMENT OF EX VIVO 2D/3D ALTERNATIVE APPROACHES for REPLACEMENT • PDX-derived culture (2D and organoids) recapitulate the heterogeneity of the patient’s tumor and have the great potential to reduce animal experimentation. • Tumor organoids are amenable to high-throughput drug screening, and can serve as a robust platform for pre-clinical pharmacogenomic studies. • Patient-derived xenograft organoids allow personalized therapy design. • Although retaining the molecular characteristics, cell cultures are not able to fully represent all biological processes and functions of the whole body including vascularization, immune cell infiltration, … • Ongoing Evaluation of ex vivo PDX-derived models in other cancer types in several laboratories across the EurOPDX member institutions. Collaborations with SMEs in the field (e.g. OcellO, The Netherlands) to define reliable cut-off parameters for drug sensitivity in PDX-derived 2D/3D cell cultures. • Development of a new platform which uses short-term cultured breast cancer PDX cells for ex vivo high-throughput single and drug-drug combination testing (Prof. Carlos Caldas lab, Cancer Research UK Cambridge Institute, in collaboration with other researchers from the EurOPDX network. Bruna et al., Cell 2016)
