WELCOME TO BIONEER International Conference on Perspectives in Precision Medicine

Transcription

1 WELCOME TO BIONEER International Conference on Perspectives in Precision Medicine Bioneer site visit March 1 th 2018

2 AGENDA Arrival at Bioneer approx Introduction to Bioneer Lovisa Sunesson, PhD, Sales & BD Manager Advanced in vitro Human Disease Models Immune models Monika Gad, PhD, R&D Manager Patient-specific Human Disease Models Bjørn Holst, PhD, R&D Manager Molecular histology Boye S. Nielsen, PhD, R&D Manager A short tour of the laboratory Departure at 15:20 - arrival at the Maersk Tower approx. 16:00 2 Bioneer site visit - International Conference on Perspectives in Precision Medicine/

3 BRIEF INTRODUCTION TO BIONEER A Danish Research & Technology Organization (RTO) - part of the Advanced Technology Group in Denmark, a so-called GTS-institute Owned by 2500 m 2 facilities; 45+ employees (30 MScs/PhDs, 12 Techs) TO 2017: ~7 mio ; 60 % commercial, 40 % R&D Key customer segments: pharma, biotech, medtech and ingredients 3 Bioneer site visit - International Conference on Perspectives in Precision Medicine/

4 BRIEF INTRODUCTION TO BIONEER, cont. Bioneer site visit - International Conference on Perspectives in Precision Medicine/

5 R&D HIGHLIGHTS Protein production Benzonase for downstream processing Mid 80 s - Hexose oxidase enzyme Identified and patented enzyme for baking Early 00 s P170 expression system Patented L. lactis protein production system 90 s-early 00 s Mammalian protein CHO/HEK cell platform 2008/2010 Oral drug models Simulated media platform 2007/2011 DGM model In-licensing of unique dynamic gut model 2013 mirna ISH Center of Excellence for Exiqon (LNA-probes) 2011/2017 ipsc technology Reprogramming of somatic cells to induced pluripotent stem cells 2014 Single-cell analysis Platform development for single cells (SNPs and mrna) 2015 RNAscope technology Certified provider for ACD on RNA-ISH 2016/ Gensplejsningsgruppen Vaccine Pig virus subunit vaccine 80 s-early 90 s Sequencing Part of Yeast genome Start 90 s Microbial diagnostics In situ detection of live bacteria Start 90 s mirna discovery Microarray and qpcr based platform 2007/2009 E-c system Effective E. coli based production platform 2009/2010 Human immune models Models that simulate critical steps in cancer and auto-immunity Gene editing Genetic engineering of ips and somatic cells 2016 Ventana Discovery Ultra Automated molecular histology platform 2016/2017 Alzheimer s disease model Full set of human ApoE isotype ipsc lines Bioneer site visit - International Conference on Perspectives in Precision Medicine/

6 WE WELCOME COLLABORATION International projects FP7 H2020 Heart disease, novel markers (6 partners/3 companies) mirna Biomarkers (6 partners/2 companies) Neuronal disease (6 partners/1 company) Cystic fibrosis treatment (7 partners/2 companies) IMI Oral delivery of biologics (23 partners/10 companies) Stem cell banking and disease models (23 partners/12 companies) Eurostars Ab purification technology (3 partners/2 companies) Immune disease (3 partners/2 companies) National projects Innovation Fund BrainStem: Stem cells in disease (4 Partners/2 companies) Infant/prenatal formula ingredients (6 partners/2 companies) Maturation of EEL (8 partners/5 companies) Cell therapy heart disease (4 partners/2 companies) 6 Bioneer site visit - International Conference on Perspectives in Precision Medicine/

7 Advanced in vitro Human Disease Models- Immune models International Conference on Perspectives in Precision Medicine Bioneer, March 1 th 2018 Monika Gad, PhD, R&D Manager, Immunology

8 Screening of drugs to meet early requirements for precision medicine In order to meet the requirements for precision medicine, screening platforms for potential drugs needs to be developed based on materials from healthy donors. Do we see the intended effect? Do we see adverse (immune toxicity) effects in general? This early approach will allow us, at a later stage, to predict more accurately which treatment for a particular disease will work in which groups of patients. Bioneer site visit - International Conference on Perspectives in Precision Medicine/

9 Bioneers immune in vitro models simulating disease conditions Bioneer has developed primary human in vitro cell assays simulating disease conditions (e.g. autoimmunity and cancer) in order to evaluate the effect of potential drug compounds. The Immune disease models are based on interactions between Dendritic cells (DC) and T-cells. The models are optimized in order to: Induce relevant cytokine profiles Induce relevant surface markers Induce relevant T-cell stimulation/proliferation, when cocultured with disease-conditioned DCs Bioneer site visit - International Conference on Perspectives in Precision Medicine/

10 Dendritic cell (DC) based Disease Models- Cytokines as Biomarkers for Immune Pathways Models mimics in vivo function under inflammatory or immune suppressed conditions

11 Generation of Monocyte Derived Dendritic Cells- Subtypes evaluated by multiple biomarkers Bioneer site visit - International Conference on Perspectives in Precision Medicine/

12 Dendritic Cell Surface (activation) markers as indicators of Immune Stimulatory Capacity C D C D 4 0 DC10 (Tolerogenic) M F I M F I M F I M F I M F I D C 1 0 D C 1 D C 1 0 D C C D P D L DC1 (Immunogenic) M F I D C 1 0 D C C D D C 1 0 D C P D L D C 1 0 D C 1 0 D C 1 0 D C 1

13 p g /m l m e a n S D p g /m l p g /m l Immunosuppressive DC (DC10)/T cell model simulating conditions seen in cancer The Mixed Lymphocyte assay as disease model Simplified immune reaction allowing study of receptor/ligand interaction IL 1 0 fr o m D C s P r o life r a tio n in M L R IF N fr o m M L R Screen for identification of single or synergistic targets (e.g. checkpoint inhibitors) D C 1 0 D C 1 0 D C 1 0 D C 1 0 D C 1 0 D C 1 DC10s shows low immune stimulatory capacity

14 C P M (3 H -T d R ) p g /m l p g /m l Pembrolizumab (Anti-PD1) can activate the immune system under immunosuppressive conditions as seen in cancer PD1: immune checkpoint molecule or inhibitory receptor leading to reduced immune effector function DC10 P r o life r a tio n in M L R IF N fr o m D C 1 0 M L R T N F fr o m D C 1 0 M L R P e m b ro liz u m a b Is o ty p e c trl P e m b ro liz u m a b Is o ty p e c trl P e m b ro liz u m a b Is o ty p e c trl m A b ( µ g /m l) m A b ( µ g /m l) m A b ( µ g /m l)

15 Summary Bioneer s in vitro immune disease models mimic critical steps in allergic, autoimmune and cancer diseases, with the aim to evaluate the effect of relevant drug compounds. This approach will allow us to predict which treatment strategies for a particular disease will work with as few adverse effects as possible In general In specific patients Bioneer site visit - International Conference on Perspectives in Precision Medicine/

16 Patient specific Human Disease Models International Conference on Perspectives in Precision Medicine- Bioneer March 1 th 2018 Bjørn Holst, PhD, R&D Manager, Disease Models

17 Precision Medicine Induced pluripotent stem cells Induced pluripotent stem cells combined with gene editing offers the possibility of designing patient specific cell models to pin-point the effect of SNPs 17 Bioneer site visit - International Conference on Perspectives in Precision Medicine/

18 Establishment of ipsc from patients and healthy individuals Gene editing Lucendo-Villarin et al

19 Gene Editing Not just ipsc lines, also imortalized cell lines Bioneer site visit - International Conference on Perspectives in Precision Medicine/

20 Neurodegenerative disease models Familiar Alzheimers disease APP PSEN1 Familiar Frontotemporal dementia MAPT GRN1 CHMP2B Risk genes in AD APOE CD33 TREM2B 20 Bioneer site visit - International Conference on Perspectives in Precision Medicine/

21 Hepatocyte differentiation

22 Organoid Hepatocyte suspension culture 96-well spheroid dish Albumin 22

23 CYP450 enzymes Cytochrome P450 enzymes are essential for the metabolism of many medications Genetic variability (polymorphism) in these enzymes may influence a patient's response to commonly prescribed drug classes CYP450 enzyme polymorphism is responsible for observed variations in drug response among patients Having access to an in vitro liver cell model expressing P450 enzymes combined with gene editing, will enable study of CYP450 SNP s in isogenic cells models Increased activity Decreased activity Absence of enzyme 23 Bioneer site visit - International Conference on Perspectives in Precision Medicine/

24 March 1 st, Precision Medicine, CPH. Bioneer - molecular histology Boye Schnack Nielsen Bioneer A/S, Hørsholm, Denmark

25 Molecular Biomarkers Nucleus Transcription microrna 5 long ncrna mrna Translation 3 UTR

26 Molecular Histology Techniques dna FISH IHC mrna ISH mir/ncrna ISH Diagnostic reagents/assays Diagnostic reagents/assays Challenging technologies, not standard in diagnostic Labs

27 RNA the world of the transcriptome trna rrna mrna lncrna microrna circrna snrna and snorna pirna and more

28 RNA detection techniques RNA sequencing / NGS microarray analyses RT-qPCR profiling NanoString profiling

29 But some biological information is missing fibrosis? vascular biology? inflammation? epithelial compartments? necrosis? apoptosis? RNA in situ hybridization may provide the answers.

30 staining label detecting probe RNA target sequence Soares et al, NAR, 2017

31 branched DNA technology / RNAscope

32 CD31/pecam1 mrna expression in colon cancers / RNAscope

33 Estrogen Receptor mrna expression in breast cancers using RNAscope Thomsen et al, unpublished data

34 Locked nucleic acid (LNA ) Center of Excellence in LNA -based situ hybridization in Certified provider of RNAscope Assays for in situ hybridization

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36 mir-21 and mir-126 in IBD, qpcr vs. qish (I) Thorlacius-Ussing et al, IBD, mir mir p<0.05 p<0.05 p<0.05 RT-qPCR Image analysis p<0.005 p<

37 Discover any RNA mrna microrna lncrna circrna. Your favorite combinations - in your favorite research tissues and cells

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39 Thanks to partners and technology platforms The Agency of Research & Innovation CarTarDis

40 THANK YOU Questions? IMMUNE MODELS Monika Gad, R&D Manager, Immunology DISEASE MODELS Bjørn Holst, R&D Manager, Disease Models MOLECULAR HISTOLOGY Boye Schnack Nielsen, R&D Manager, Mol. Histology MOLECULAR DETECTION Kim Holmstrøm, R&D Manager, Molecular Detection BUSINESS DEVELOPMENT Lovisa M. Sunesson, Sales & Business Dev. Manager 24