Presented by Dr Malcolm Haddrick on 10th June 2020

1. WHO WHY HOW WHAT WHERE WORK WITH US
Medicines Discovery Catapult Connects Webinar
Wednesday 10th June 2020
Dr Malcolm Haddrick
Lead Scientist
MDC
Challenges and Opportunities of Complex
Cell Models for Toxicity Testing

2. WHO WHY HOW WHAT WHERE WORK WITH US
Complex Cell models for toxicity:
Opportunities and Challenges
• Standard Cell Culture, simple models
• Immortalised
• Single Cell Type
• 2D
• Limited biological stimuli
• Limited physical stimuli
• Limited translation to patients
• Targets for toxicity
• Absence of a ‘system’
• Next Generation Models
• Stem, Primary, CRISPr
• Multiple Cell Types
• 3D, Organoids
• Mimic biological environment
• Accommodate flow or ECM
• Improved translation to patients
• Targets for toxicity
• Limited ‘system’
What’s the problem ? What’s the solution ?

3. WHO WHY HOW WHAT WHERE WORK WITH US
Develop and enable access to well validated humanised isolated and connected
(Organ on a chip, OOAC) cell models for hypothesis testing, efficacy and safety
determination meeting the needs of UK Drug Discovery
Bring industrial experience to exhaustively characterise the cell model
system, it’s robustness and it’s limitations to exemplify use. Working
collaboratively
Establish on-chip solutions for single and multi-organ models. Prototype new
technology. Impact NC3Rs
Implement increasing complex models

4. CMEF Cardiac Model
WHO WHY HOW WHAT WHERE WORK WITH US
Establish a 3D cardiac model for assessment of cardiotoxicity
Day 4 Day 19
• iPSC derived cardiomyocytes with primary cardiac fibroblasts &
endothelial cells (CMEF)
• Spheroid formation in ULA plates, spontaneous beating from ~ 1 week
• Determination of beat rate and amplitude on the electrode ofa
Cardioexcyte instrument

5. CMEF Cardiac Model
WHO WHY HOW WHAT WHERE WORK WITH US
Day 4
• Exemplar data for characterisation on CE96
• Off chip advanced cell model – predictivity with mixed cell types.
Beat rate & amplitude
Electrical activity
Spontaneous beating3D cardiac microtissue
Day 19
Pre cpd
Post cpd
Dose
dependent
Dofetilide
effect
T2T
Cardiac
hypertrophy

6. WHO WHY HOW WHAT WHERE WORK WITH US
CMEF Predictivity with known human cardiotoxins
Ravenscroft et
al., Tox Sci
152(1), pg 99-
112.
• Validation of the model using known clinically relevant human cardiotoxins
• Improved predictivity as a consequence of multiple cell types, e.g. detection of dobutamine
• Amenable to screening at discovery scale, label free, continuous and real time measurements
• Ongoing collaborations for CRISPr introduced cardiac mutations and endothelial cell optimisation

7. Blood Brain Barrier Model- The
opportunity
• BBB – Blood vessels of the CNS that regulate the movement of molecules,
ions. Consists of endothelial cells lining the vessels held together by tight
junctions, surrounded by astrocytes and pericytes within a matrix
environment
• Current standard in vitro BBB models use Caco-2 or MDCK-MDR1 cell lines in
transwell plates along side in vivo rodent models for compound permeability
and neurotoxicity
• Need for more physiologically representative and human relevant in vitro
blood brain barrier models

8. Blood Brain Barrier Model - OOAC
● Mimetas OOAC- 384 well plate SBS format
● 96-chip or 40-chip format, ability to form a tubule, simple flow
● Compartmentalised channels, allows cross talk between cells
● primary endos, pericytes, astrocytes and investigation of iPSC sources
Endothelial
tubule

9. BBB On Chip Tubule formation
Primary human Endothelial cells
(hBMECs)
Cell Source
Tubule Formation
Viability & Cell health
Barrier integrity & permeability
DAPI Actin DAPI Actin
Tight Leaky

10. Liver Models
WHO WHY HOW WHAT WHERE WORK WITH US
• Primary hepatocytes need flow to maintain ex vivo functionality
• Physiomimix OOAC platform from CN Bio Innovations
• Primary hepatocytes remain viable and functional (urea, albumin, CYP)
Day 4 Day 19

11. Liver Models – NASH/NAFLD
WHO WHY HOW WHAT WHERE WORK WITH US
• Hepatocytes, stellate and Kupffer cells co-culture and fat loaded to establish a NASH/NAFLD model
• Ability to determine hepatotoxicity in a ‘diseased’ human model
aSMA Col I DAPI
• 3D channel containing the 3 cell types

12. WHO WHY HOW WHAT WHERE WORK WITH US
• Multiple providers, enabling model viability and biological communication. Excitement and hype !
OOAC – connecting multiple cell models
Edington et al., Nat Sci Rep 2018 8:4530
• A connected cardiac & liver model
• Reactive metabolite mediated toxicity
• Generated in liver, toxic to heart
• 2, 3, 4, 7 connected chambers…..
• Towards human on a chip
• For investigational toxicity, can these systems ever be good enough to cover all targets & systems ?

13. WHO WHY HOW WHAT WHERE WORK WITH US
Complex Cell Models – Challenges
• Isolated on chip models
• Definition
• Complex enough ?
• What is it
replacing ?
• What are the
limitations ?
• Validation
• Connected on chip models
• Connected organs
• Deficiencies
• Relevance of
communication
• Missing organs
• Coverage of targets
for toxicity
• Translation to man
• How to derive
• Scaling
• Comparable data
• Predictivity
• 3Rs benefits
• Reason to believe
• Reason to invest $
• Regulatory ?
• Measurements
• Label / label free
• Real time
• Continuous, end pt
• Viability
• MS, genomics
• Compound effects
• Market – demand
• Tech & Biology
• Equipment
• Service / CRO
Community innovation, share issues and promote adoption