This document summarizes research from the Thorax Institute on using induced pluripotent stem cells (iPSCs) to model metabolic diseases like familial hypercholesterolemia (FH). Researchers generated iPSCs from a FH patient and differentiated them into hepatocyte-like cells. These cells reproduced the patient's phenotype of impaired LDL cholesterol uptake and responded to pharmacological treatment. The researchers propose that 3D culture systems could further mature the cells to enable phenotypic screening of diseases like non-alcoholic steatohepatitis and fibrosis.

1. BANDEAU TUTELLES
L’unité de recherche de l’institut du thorax
Inserm UMR 1087 / CNRS UMR 6291
Nantes, France
3D innovative hiPSC-based models including the
microenvironment
for phenotypic screening - Application on metabolic
diseases
Karim Si-Tayeb
ELRIGfr Conference 23/24 mai 2018 Le Bischenberg, Bischoffsheim, France

2. 2
Familial hypercholesterolemia (FH)
Facteurs de risque génétique
Récepteur
LDL
Inconnus
APOB
PCSK9
Hypercholesterolemia:
Impaired LDL-cholesterol uptake
Modified from Norata et al. EHJ 2013
Identification of informative famillies displaying:
- A spontaneously low LDL-C level (FHBL)
- A hypercolesterolemia without cardiovascular disease
(CIC Thorax, Matthieu Pichelin, Marie Marrec)
Absence of mutations in known genes involved in FH
(LDLR, PCSK9, APOB, ANGPTL3)
(Jocelyne Magré, Jean-Jacques Schott)

3. 3
The Scientist Magazine
April 2016
Si-Tayeb et al. Disease Models & Mechanisms 2016
Patient
Urine-derived cells
hiPS cells
in vitro study of familial hypercholesterolemia
Hepatocytes
© l’institut du thorax
Modeling FH in vitro from an urine sample - the power of hiPS cells

4. 4
From hiPS cells toward hepatocytes – back to
developmental biology
He: precardiac mesoderm
End: endoderm
Lb: liver bud
E: endothelial cells
STM: Septum transversum
d0 d5 d10 d15 d20
Hepatocytes
Definitif
Endoderm
Hepatic
progenitors
Specified
Definitif
Endoderm
Human
pluripotent
stem cells
Si-Tayeb et al. Developmental Cell 2010
Si-Tayeb et al. Hepatology 2010

5. 5
Fluorescent LDL uptake analyzed by flow cytometry
Phenotype
reproduced in vitro
Familial hypercholesterolemia in a petri-dish
Phenotype
pharmacologicaly
reversed in vitro
Si-Tayeb, Idriss et al. Disease Models & Mechanims 2016

6. Hypercholestérolémie:
Défaut d’internalisation du LDL-choléstérol

7. 7
A promising model that needs further maturation by adding a third dimension
Nature review 2004

8. 8
Liver
A promising model that needs further maturation by adding a third dimension
Amandine Caillaud Méryl Roudaut

9. 9
A new differentiation protocol for a new environment
Cytokines
d0 d5 d10 d15 d20
Hepatocytes
Definitif
Endoderm
Hepatic
progenitors
Specified
Definitif
Endoderm
Human
pluripotent
stem cells
Méryl Roudaut

10. 10
CYP450 basal activities and induction
CYP450 2C9 (ibuprofen)
Diclofenac -> 4OH-Diclofenac
CYP450 1A2 (caffein)
Phenacetin-> Acetaminophen
CYP450 2A6
Nicotine-> Cotinine
CYP450 2E1 (ethanol)
Chlorzoxazone-> 6-0H-Chlorzoxazone
Méryl Roudaut
(nM)
(nM)
(nM)(nM)

11. 11
CYP450 basal activities and induction
CYP450 3A4
Testosteron-> 6ß-OH-testosteron
0
100
200
300
400
500
0 10 20 30 40 50 60 70
nmol/L
Minutes
CYP3A4 activity (nmol/L/min/well)
Basal activity
Induced activity
0
50
100
150
200
250
Basal activity Induced activityActivitypercentage
CYP3A4 activity
*
Méryl Roudaut
(nM)

12. 12
Conclusion and perspectives
hiPS cells -> Genetic variability
Hepatic cells -> Phenotypic screening
3D
2D
6 puits 96 puits

13. 13
Conclusion and perspectives
Liver NASH Fibrosis

14. www.umr1087@univ-nantes.fr
L’unité de recherche de l’institut du thorax
Inserm UMR 1087 / CNRS UMR 6291
Nantes, France
Daniel Buren et Patrick Bouchain, Les Anneaux, Quai des
Antilles, Nantes, création pérenne Estuaire 2007 © Martin
Argyroglo/LVAN
L’institut du thorax
CARIOU Bertrand
LE MAY Cédric
MAGRE Jocelyne
PRIEUR Xavier
PICHELIN Matthieu
ARNAUD Lucie
AYER Audrey
BLANCHARD Claire
Wargny Matthieu
MOREAU François
CHADEUF Gilliane
DIJK Wienecke
THEDREZ Aurélie
SI-TAYEB Karim
IDRISS Salam
CAILLAUD Amandine
GIRARDEAU Aurore
ROUDAUT Méryl
CANTAT Pierre-Alexandre
LORANT Victoria
HCS Pharma
MAUBON Nathalie
MAUBON Gregory
SOUGUIR Zied
VANDENHAUTE Elodie