Abstract : a single procedure to generate functional hi ps cs-derived liver organoids - towards an innovative tools suitable for drug screening

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HCS Pharma

Abstract for poster for the "VIIIème Colloque de Génomique Fonctionnelle du Foie" - Rennes (France) 2020

Santé & Médecine

VIII
ème
Colloque
de Génomique Fonctionnelle du Foie
A single procedure to generate functional hiPSCs-derived liver
organoids - Towards an innovative tools suitable for drug screening
Méryl Roudaut 1, 2, Amandine Caillaud 1, Aurélie Thédrez 1, Wieneke Dijk 1,
Aurore Girardeau 1, Matthieu Pichelin 1, Lucie Arnaud 1, Mikaël Croyal 1, 3, Cédric
Le May 1, Élodie Vandenhaute2, Zied Souguir 2, Nathalie Maubon 2, Bertrand
Cariou 1, Karim Si-Tayeb 1
1°) Institut du thorax, INSERM U1087, CNRS 6291, IRS-UN, CHU-Nantes, Nantes, France
2°) HCS Pharma, Lille, France
3°) Plateforme de Spectrométrie de Masse, CRNHO, Inra UMR 1280, Nantes, France
We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable
model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. With a
non-invasive approach, hiPSCs can be generated from urine samples of patients and HLCs
have been used to model cholesterol metabolism regulation, by the study of LDLR- and
PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-
mediated familial hypobetalipoproteinemia (FHBL). This model provides promising
advantages with a direct link to the patient and with an unlimited source of HLCs. But like
all models, there are limitations, mainly by the neonatal characteristic of HLCs lead to
difficulties for pharmacological investigations.
Therefore, to overcome these burdens, we chose to 1. Differentiate hiPSCs into HLCs in
an innovative 3D modified Hyaluronic Acid hydroscaffold, BIOMIMESYS® produces by HCS
Pharma to enhance their maturation. 2. Adapt our 3D differentiation process to a 96-well
format to make it compatible for drug screening. 3. Characterization of the 3D HLCs
model by metabolism tests and compare to primary human hepatocyte (PHH).
We gathered 3’ SRP data all along the differentiation process and RNAseq has been
performed by comparing 2D and 3D differentiation conditions to characterize hiPSCs
differentiation into liver organoïds. We observed an enhanced expression of most hepatic
genes and genes expressed by non-parenchymal cells such as stellate cells.
Immunofluorescence data confirmed the co-localization of albumin-positive hepatocytes,
desmin-positive stellate cells and LYVE1-positive endothelial cells in liver organoïds.
Finally, at a functional level, several CYP activities including CYP3A4 were detected at the
basal level and successfully induced. Liver organoïds responded to pharmacological
treatments as shown by their ability to accumulate lipids upon amiodarone treatment or
uptake LDL-bodipy upon statin treatment.
Altogether, our development gave rise to functional liver organoïds generated with a
unique and common procedure, in a process of automating for future high throughput
screening.
E-mail correspondant/corresponding author : karim.si-tayeb@univ-nantes.fr

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