MDC Connects: Go native… Characterising therapeutic effect in primary cellular models
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This document discusses the advantages of using primary cellular models in drug development. Primary cells better mimic native biology and target expression compared to cell lines. Several case studies are presented where primary cellular models helped validate drug targets and identify potential safety issues prior to clinical trials. The document emphasizes characterizing fit-for-purpose models to generate relevant data at each stage of development from discovery to clinical translation. Thorough assay development and validation are important to support clinical applications.
MDC Connects: Go native… Characterising therapeutic effect in primary cellular models
- 1. Go Native … Amanda Woodrooffe 03 June, 2020 Characterising therapeutic effect in primary cellular models
- 2. 2 ▪ Target validation – where do cellular models fit ? ▪ Primary cells – advantages, disadvantages, opportunities & positioning ▪ Case Studies ▪ Why go native ? ▪ The importance of assay development, characterisation and appropriate validation ▪ Applications from discovery to clinical – support translation Overview Going native…
- 3. 3 ▪ Attrition during clinical development remains an issue ▪ Often attributed to inadequate pre-clinical target validation and/or suitable clinical translation data ▪ Development and application of relevant preclinical cell based models can help ! ▪ The most important considerations: ▪ What is your question ? ▪ What are the relevant data to enable progress ? ▪ How to characterise the relevant biology ? ▪ What model is most fit-for-purpose ? ▪ Complexity of disease: involvement of different cells ? Target Validation Use of cellular models There Remains A Need To Address Clinical Attrition and Efficacy Challenges • Thorough understanding of complex disease and target biology • Better human / clinical predictive models and biomarkers
- 4. 4 ▪ Advantages ▪ Most “in vivo like” ▪ Native target(s) ▪ Interindividual variation ▪ Plethora of culture formats / media Going Native : primary cells When, Why, How ? ▪ Disadvantages ▪ Availability / sourcing ▪ Scale ▪ Interindividual variation ▪ Maintenance of cellular phenotype ▪ Positioning ▪ Tertiary screen / candidate selection ▪ Higher throughput screen (e.g. PBMCs) ▪ Clinical translation / biomarker(s) ▪ Opportunities ▪ High volume isolations ▪ Cryopreservation ▪ Pooling donors Human bronchial epithelial cells (2D) Human bronchial epithelial cells (3D) Human stellate cells (2D) Human liver (3D co culture)
- 5. 5 ▪ Increase in intracellular free Ca2+ in response to acetylcholine in NLEC and HLE-B3 cell line ▪ Consistent in cell line and native cells Case Study 1 : Be more in vivo like - cell line vs native Is it the right biology ? From Collison et al., Invest Ophthalmol Vis Sci. 2000 0 25 50 75 100 125 0.001 0.01 0.1 1 10 Responseamplitude(%) Antagonist concentration (uM) 0 25 50 75 100 125 0.001 0.01 0.1 1 10 Responseamplitude(%) Antagonist concentration (uM) ▪ But, responses are antagonised differently ◆ Pirenzipine (M1 antagonist) AF-DX384 (M3 antagonist) Native cells - NHLEC Cell line - HLE-B3 0 2000 4000 6000 8000 M1 M2 M3 M4 M5 Muscarinic sub-type Copynumber 0 500 1000 1500 2000 M1 M2 M3 M4 M5 Muscarinic sub-type Copynumber M1 M2 M3 M4 M5 M1 M2 M3 M4 M5 0 0.25 0.5 0.75 1 1.25 0.001 0.01 0.1 1 10 100 1000 RelativePeakAmplitude Agonist Concentration (uM) NLEC HLE-B3
- 6. 6 ▪ Model : Recapitulates native islet morphology in prolonged culture ▪ Target(s) : Maintenance of key pharmacological targets e.g. GPR40 ▪ Relevant function(s) : GSIS & responsive to standard secretagogues ▪ Reliable : Robust, reproducible methodology – data from over 80 individual donors Assay development and characterisation islet ORGANDOT 3D culture Pancreatic islets in situ islet ORGANDOT cultures Insulin + ProInsulin Insulin + C peptide Insulin + Glucagon Insulin + Somatostatin Insulin GLUT2 glucokinase GLP1 R GPR40 GPR119 glucagon
- 7. 7 ▪ A potent and selective GPR40 agonist is theorized to be a safe and effective antidiabetic drug with little or no risk of hypoglycaemia. ▪ TAK-875 (Fasiglifam) was effective at potentiating insulin secretion in islet ORGANDOT cultures ▪ TAK-875 was withdrawn from Phase III clinical development due to drug-induced liver injury (DILI) ▪ Retrospective analysis in BioIVT’s C-DILI model demonstrated the hepatic safety issues observed clinically ▪ Effective use of primary cellular models, in combination, to validate target, evaluate drug effect & potential safety liability Case Study 2 : GPR40 agonist for diabetes Getting the balance right… islet ORGANDOT data C-DILI data (courtesy of BioIVT)
- 8. 8 Case Study 3 : From discovery to clinical Development and full validation to support Marketing Authorisation Application ▪ Human primary cell-based assay for screening anti-TNFa biosimilars ▪ 2D, synovial fibroblasts, RA donors ▪ Assay development ▪ Reference standard inhibitor ▪ Titration of stimulus ▪ Selection of sentinel marker ▪ Assay validation ▪ Linearity and range ▪ Relative potency ▪ Parallelism ▪ Specificity – formulation effect ▪ Precision – intra / inter assay ▪ Formulation effect ▪ Biosimilar screen ▪ Relative potency versus Humira Assay being used to screen small molecules in discovery too…
- 9. 9 ✓ Modelling native biology at native receptors in simple or complex cellular systems can be achieved ✓ Positioning of your assay – consider format, complexity, throughput, source materials – suitable for secondary or tertiary screen ? ✓ Characterise a fit for purpose model – what actionable data are required ? Make sure your model delivers. ✓ Level of validation needed – multiplexed endpoints, clinically relevant, triage to selected clinical markers ? ✓ Consider the bigger picture – efficacy and safety… Summary Keep thinking about clinical translation…
- 10. THANK YOU! we look forward to staying in touch Credit: Patrick Foley, Cheyenne, Wyoming, USA From: The Pathologist, April 2020 Keep those primary cells smiling… ! amanda.woodrooffe@precisionformedicine.com