new approaches to establishing safety and conducting .../media/files...new approaches to...
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New Approaches to Establishing Safety and Conducting Toxicology Studies
Thomas Hartung & team
Center for Alternatives to Animal Testing
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Co-founder - organoids
Consultant Computational Toxicology
Green Chemistry Advisory Panel
Licensed Pyrogen Test Consultant
Conflict of Interest Statement
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• 20% drug failure because of toxicity not predicted
• 8% drugs withdrawn from market because of side-effects
• Occasional problems first in humans
• Information too late in process • Poor prediction of drug
interactions (killing 1:100 hospitalized patients)
• Sorting out of substances because of meaningless (animal and mutagenicity) tests
We are
not 70kg
rats !!!
Some problems of toxicology in pharma
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Toxicology has the likely most reproducible animal tests (GLP, standardized protocols, some validated, high doses, substance effects in healthy animals) – their reproducibility:
• Cancer bioassay: 57% (repeat or mouse vs. rat) • Reproductive Tox: 60% between species • Uterotrophic assay: 26% contradictory • Skin sensitization: 77% guinea pig vs. mouse • Severs eye irritation: 70% reproducible • Acute fish tox: up to 6 log orders different • Chronic tox: no correlation between
mouse and rats or genders
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Irreprodu-cell-bility Cell tests have not less problems!
• Ca. 25% of cell lines misidentified • 15-25% mycoplasma infected • Genetic instability • Culture artifacts
Pronounced genetic differences in frozen cells of the same lot from a cell bank Kleensang (2016) Sci Rep 6, 28994
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Workshops 2015
Jun, Baltimore
Dec, Konstanz
International Good Cell
Culture Practice
Collaboration
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Curated Legacy Data Read-Across
(Multi-)Omics techologies
High-Throughput Screening
High-Content Imaging
The changing approach in toxicology
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Mapping the Human
Toxome by Systems
Toxicology
In vitro model
omics data generation
Pathways of Toxicity
Software tools
Validation tools
Human Toxome
Database
NIH Transformative Research Grant 15 publications, 2 under review http://humantoxome.com
Pathways of toxicology from multi-omics
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Mining REACH data made machine-readable by natural language processing
Largest toxicological data-base = “goldmine” for computational toxicology
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11 Science, 12 Feb 2016
CAAT Read-Across Collaboration • Inter national, cross-industry and
multi-stakeholder steering group • One workshop and two symposia;
2017 regulator summit • Good Read-Across Practice and
guidance on biological support data
• Application to Green Toxicology (frontloading of toxicology)
• Automated tool development • Collaboration with UL making
tool and services available • Expansion to other data-bases
including pharma • Additional uses e.g. drug
contaminants
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http://en.wikipedia.org/wiki/Organ-on-a-chip Stem cells &
Organo-typic
culture & High-
content
21st century
science starts with
21st century cell
culture
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Concept articles, workshops and symposia Transatlantic Think Tank for Toxicology
Promotion of organotypic cultures and
Integrated testing Strategies
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Our mini-brain project
• Stable size, no necrosis • Stable composition (10%
oligodendrocytes, 20% astrocytes, 5+ types of neurons)
• 40% myelinated axons • Electrophysiologically active
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Announced at AAAS 12
Feb 2016
80+ articles
Huge expectations
Some quote me
correctly:
But, right now, he
cautions, “I’m selling
hopes,” as the mini-
brains are brand new
and haven't proven
useful yet.
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Meperidine addiction led to Parkinson via MPTP contamination
Standard animal model Similar: pesticide Rotenone
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Parkinson model
MPTP and Rotenone selectively kill dopaminergic neurons
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Diseases like autism cannot be
explained by genetics or exposure alone
Disposition to
DNToxicants? Test on a genetic
background that allows DNT
0.00
10.00
20.00
30.00
40.00
10 100 10000
20
40
60
80
100
10 100 1,000
CHD8+/- CHD8+/+
Chlorpyrifos,μM
NPC 8weekorganoids
%cytotoxicity
%cytotoxicity
A B
Synergy of
CHD8 and
chlorpyrifos,
i.e. two ASD
risk factors
ALTEX 2014, 31:129-156
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Use for viral infection Examples: JC virus Zika HIV
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Goals: • Not Brainome but
Organome • Get Organoids out
of academia to practical use
• Help standardization, QA and validation
• Make visible
Nature Medicine 6 Apr 2016
http://www.organome.co
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NF/TH/DAPI NF/TH/DAPI
Control Frozen
Contr
ol
Froze
n
0
100
200
300
Mitotracker
condition
Me
an
gra
y v
alu
e
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Cryopreservation of mini-brains - Neurite outgrowth -
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Academic model in cell incubator
Weekly production 3 month model
Several cell lines continuous production
Stockpiling of frozen mini-brains
The advantage of “brain on the rocks”
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Get frozen mini-brains where they are needed
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Are our cell cultures good enough?
?
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I cannot say whether things will get better if we change; what I can say is they must change if they are to get better. Georg Christoph Lichtenberg (1742 - 1799)