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HCS for brain disorders / HCS Pharma at B4B mars 2018

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How phenotypic screening with innovative cellular models can help to unravel mechanisms or repurpose drugs in the frame of brain disorders

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HCS for brain disorders / HCS Pharma at B4B mars 2018

  1. 1. www.hcs-pharma.com Biotechnology company specialized in cellular imagery which provides HCA/HCS services How phenotypic screening with innovative cellular models can help to unravel mechanisms or repurpose drugs in the frame of brain disorders (?) Friday, March 9th Elodie VANDENHAUTE, PhD
  2. 2. www.hcs-pharma.com 2 Drug discovery process 10-16 years 300k - 1 million coompounds 1 drug
  3. 3. www.hcs-pharma.com 3 Approaches for identifying new drugs in R&D processes: target-based screening High-Throughput Screening (HTS) = method of experimentation comprising the screening of large compound libraries for activity against defined biological targets via the use of automation, miniaturized assays, and large-scale data analysis. Source: Bellomo et al., 2017 in J Inherit Metab Dis - Much data very rapidly - One target/parameter at a time - Target out of its cellular context - Some targets not known… - Low cost
  4. 4. www.hcs-pharma.com Approaches for identifying new drugs in R&D processes: phenotypic screening Source: Bellomo et al., 2017 in J Inherit Metab Dis Phenotypic Screening = testing a large number of compounds in a target-agnostic assay that monitor phenotypic changes in animals or cells 4 High-Content Screening (HCS) = method to automatically screen and analyse the temporal and spatial activities and functions of cells and their constituents using microscopy - Time-consuming - High storage capacities - Multi-parametric - Cellular context - Target unknown - Single-cell level - Unexpected discoveries !
  5. 5. www.hcs-pharma.com High content screening: principle 5 96- or 384-well plates
  6. 6. www.hcs-pharma.com What can be studied with HCS at the cellular/subcellular level? 6 Blue: Hoechst ; Green: GAPDH ; Yellow: Il-8 ; inducer: TNFα Cells : HeLa ; Affymetrix ViewRNA ISH assay kit mRNA analysis Blue: Hoechst ; Yellow: AQP3 Cells: NHEK Blue: Hoechst ; yellow: 53BP1 ; red: γH2AX ; Cells: NHEK Protein expression and localization Blue: Hoechst ; red: ROS ; Cells: NHEK A) Menadione B) Menadion e + NAC A B Stress: ROS, mitochondrial potential … Blue: Hoechst Cells: HeLa Cell number, cell cycle analysis Blue: Hoechst ; red: EdU Cells: NHEK Cell number, proliferation Proliferation, apoptosis, differentiation Blue: Hoechst ; Green: Neutral Lipids Cells: HepaRG Blue: Hoechst ; Red: ROS Yellow: mitochondria pot Cells: NHEK Vacuoles/vesicles Cell structure : organelles, cytosquelette, … Cell motility : tracking (video)
  7. 7. www.hcs-pharma.com Example of our high content screening platform 7 Incubator 37°C/CO2 Storage 4°C Plate washer & sealer (behind) Pipetting platform with 96/384-well head Plate hotel (RT) 2 ImageXpress Micro confocal automated imaging systems (MolDev) Platform integrated by + Server
  8. 8. www.hcs-pharma.com 8 How HCS could help to fight brain diseases? By finding ‘new’ drugs
  9. 9. www.hcs-pharma.com HCS for finding drugs against brain diseases? 9 How to shorten the timeline for drug discovery and reduce costs? 10-16 years 300k - 1 million coompounds 1 drug Find new applications for existing drugs = drug repositioning Ref: Rossi et al., 2016 in Dermatol Ther Until recently, drug repositioning < accidental observations, known side effects, mechanisms of action of individual compounds. Ex: minoxidil = antihypertensive drug Observation: ↑ hair growth Now used to treat androgenic alopecia  HCS = new way to identify already approved drugs to be repurposed against rare diseases, in unbiased experimental settings
  10. 10. www.hcs-pharma.com HCS for repurposing drugs: example of Zika virus infection and induced neural cell death 10Ref: Xu et al., 2016 in Nature Medecine Clinical observations of ZIKV in fetal brains (infected women): Zika Virus efficiently targets human neural progenitor cells (hNPCs) and attenuate their growth. Cell cultures: hNPCs and human astrocytes derived from iPSCs + glioblastoma SNB-19 cells
  11. 11. www.hcs-pharma.com HCS for repurposing drugs: example of Zika virus infection and induced neural cell death 11Ref: Xu et al., 2016 in Nature Medecine  Emricasan suppresses cell death of ZIKV-infected human astrocytes and hNPCs in 2D monolayer cultures and in 3D brain organoids. 2D 3D
  12. 12. www.hcs-pharma.com 12 How HCS could help to fight brain diseases? By unraveling mechanisms of diseases (finding new therapeutical targets)
  13. 13. www.hcs-pharma.com HCS for unraveling mechanisms of diseases? Example of high-risk neuroblastoma (NBL) 13 Imaging-based RNAi screen for proliferation and differentiation of NBL cells Ref: Veschi et al., 2017 in Cancer Cell SETD8 (KMT5A) encodes lysine- methyltransferase enzyme
  14. 14. www.hcs-pharma.com HCS for unraveling mechanisms of diseases: example of high-risk NBL 14 Effects of in vitro treatment of SK-N-AS cells with an inhibitor of SETD8 (UNC0379) Ref: Veschi et al., 2017 in Cancer Cell  Epigenetic-focused HCS RNAi (+ chemical) screens identified SETD8 as an essential factor for NBL survival and maintenance in an undifferentiated state.
  15. 15. www.hcs-pharma.com Importance of the cellular models in HCS 15 Basis of HCS technology = cellular models - Cell lines - Primary cell cultures - Cells derived from stem cells (progenitor cells, or induced pluripotent stem cells) Nature? Origin? - Animal cells (drosophila, mammalian etc.) - Human cells Complexity? - Monocultures in 2D/3D - Cocultures in 2D/3D Predictability?
  16. 16. www.hcs-pharma.com How HCS can help to fight LSDs? 16 • By finding efficient drugs though repositioning • By finding new/relevant cellular targets (better understanding underlying mechanisms) • Benefits of new cellular models ? iPSCs ?
  17. 17. www.hcs-pharma.com Our ongoing projects (CNS) 17 The blood-brain barrier (BBB) 1. Miniaturization (12 puits  96 puits) 2. Combination with neuronal and glial cells, to consider the BBB in toxicity and efficiency tests (healthy + pathological models) ? . . . . . ...... .. . . . . PhD grant (CIFRE, Véronique De Conto)
  18. 18. www.hcs-pharma.com BIOMIMESYS®: a natural HA-based hydroscaffold to better mimic the microenvironment (ECM) 18 BIOMIMESYS® Adipose tissue Decellularized human adipose tissue BIOMIMESYS® Liver Decellularized liver ECM BIOMIMESYS® Brain under development BIOMIMESYS® is biofunctionalized with other matrix components (collagens, etc.) and ready-to-use in cell culture plates. 100µm 96-well plate with lyophilised matrix Differentiated Luhmes cells
  19. 19. www.hcs-pharma.com Our scientific advisory board and academic collaborations 19 Pr Romeo Ceccheli, PhD BBB expert Pr Bertrand Cariou, M.D-Ph.D Team leader RHU CHOPIN program Dr Georg Haase & Pr David Devos Neurobiologists Pr Christophe Lefebvre, PhD Scientist expert on microglia Karim Si-Tayeb, PhD Scientist in Institut du Thorax iPSC & metabolic diseases expert Pr Dominique Collard, Ph.D SMILL-E Team leader BIOMEMS expert Dr Fabrice SONCIN, DR Angiogenesis expert
  20. 20. www.hcs-pharma.com HCS Pharma team 2020 CEO, CSO Nathalie MAUBON PhD in biology Ph.D Student in metabolic diseases Méryl ROUDAUT MSc in toxicology Study director Pierre-Jean FERRON PhD in toxicology Technician in cellular biology Stéphanie MOLEZ Study director Elodie VANDENHAUTE PhD in neurobiology and physiology Cell culture leader Kathleen JARNOUEN MSc in cell biology Technician Marie Lesaffre MSc in cell biology Ph.D Student (BBB modelling) Véronique De Conto MSc in cell biology Business Developer Nathalie ACCORNERO PhD in molecular & cellular biology CDO, CIO Grégory MAUBON PhD in astrophysics Sr researcher biomaterial Zied SOUGUIR PhD in biomaterial
  21. 21. www.hcs-pharma.com Biotechnology company specialized in cellular imagery which provides HCA/HCS services Thank you for your attention ! 21

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