HCS Pharma is a biotechnology company focused on high-content screening (HCS) and cellular imagery to facilitate drug discovery and repurposing, especially for brain disorders. Their innovative methods allow for the identification of new drug applications and mechanisms of diseases through the analysis of cellular and subcellular activities. The company aims to shorten drug discovery timelines and reduce costs while utilizing advanced cellular models and technologies.

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

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Drug discovery process
10-16 years
300k - 1 million
coompounds
1 drug

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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

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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
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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 !

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High content screening: principle
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96- or 384-well plates

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What can be studied with HCS at the
cellular/subcellular level?
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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)

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Example of our high content screening
platform
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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

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How HCS could help to fight brain
diseases?
By finding ‘new’ drugs

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HCS for finding drugs against brain
diseases?
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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

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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

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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

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How HCS could help to fight brain
diseases?
By unraveling mechanisms of diseases (finding
new therapeutical targets)

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HCS for unraveling mechanisms of diseases?
Example of high-risk neuroblastoma (NBL)
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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

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HCS for unraveling mechanisms of
diseases: example of high-risk NBL
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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.

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Importance of the cellular models in HCS
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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?

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How HCS can help to fight LSDs?
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• By finding efficient drugs though repositioning
• By finding new/relevant cellular targets (better understanding
underlying mechanisms)
• Benefits of new cellular models ? iPSCs ?

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Our ongoing projects (CNS)
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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)

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BIOMIMESYS®: a natural HA-based hydroscaffold to
better mimic the microenvironment (ECM)
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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

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Our scientific advisory board and academic
collaborations
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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

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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. www.hcs-pharma.com
Biotechnology company specialized in cellular imagery
which provides HCA/HCS services
Thank you for your attention !
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