Successfully reported this slideshow.
We use your LinkedIn profile and activity data to personalize ads and to show you more relevant ads. You can change your ad preferences anytime.
1.  L'institut du thorax, Inserm UMR1087 – CNRS UMR 6291, Université de Nantes, Nantes, France.
2.  HCS Pharma, Lille. 3. ...
Upcoming SlideShare
Loading in …5
×

Poster – Development and automation of 3D innovative hiPSC-based liver organoids including the microenvironment for phenotypic screening - Application on metabolic diseases

268 views

Published on

We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.

Published in: Health & Medicine
  • Be the first to comment

  • Be the first to like this

Poster – Development and automation of 3D innovative hiPSC-based liver organoids including the microenvironment for phenotypic screening - Application on metabolic diseases

  1. 1. 1.  L'institut du thorax, Inserm UMR1087 – CNRS UMR 6291, Université de Nantes, Nantes, France. 2.  HCS Pharma, Lille. 3. Plateforme de Spectrométrie de Masse, CRNHO, Inra UMR 1280, Nantes France Development and automation of 3D innovative hiPSC-based liver organoids including the microenvironment for phenotypic screening - Application on metabolic diseases Méryl Roudaut1,2, Amandine Caillaud1, Aurélie Thédrez1, Wieneke Dijk1, Aurore Girardeau1, Matthieu Pichelin1, Lucie Arnaud1, Mikaël Croyal3, Cédric Le May1, Elodie Vandenhaute2, Zied Souguir2, Nathalie Maubon2, Bertrand Cariou1, Karim Si-Tayeb1 INTRODUCTION We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations. Therefore, to overcome these burdens, we choose to : 1.  Differentiate hiPSCs into HLCs in a 3D environment to enhance their maturation; 2.  Adapt our 3D differentiation process to a 96 wells format to make it compatible for drug screening. CONCLUSIONS Karim SI-TAYEB, L’unité de recherche de l’institut du thorax karim.si-tayeb@univ-nantes.fr + 33 (2) 28080176 Comparaison between 2D and 3D hepatic differentiation by RNAseq www.umr1087.univ-nantes.fr A 3D environment for hiPS cells differentiation Liver Porosity & stiffness ECM compounds (Zanger and Schwab, 2013) (Brunton L and al., 2018) Testosteron disappearence 6ß-OH-Testosteron apparition Functional test - Cytochrome P450 activities and induction of hiPSC-derived liver organoids(MS) LiverBile Fatty acid CholesterolLipoprotein GlucoseInsulin CytochromeXenobiotic Hepatocyte ADH Hepatocyte FHBL Control Control ADH FHBL A newly designed differentiation protocol Undifferentiated hiPScells Hepatocyte differentiation 2D3D2D3D Undifferentiated hiPS cells Hepatocyte differentiation 2D3D2D3D Hepatocyte differentiation Undifferentiated hiPS cells 2D3D 2D3D 2D3D Volcano plot 3D hepatocytes (red) vs 2D hepatocytes (green) Benjamini & Hochberg method (7448 /57905 DE genes) Log2(fold change) CYP1A2 (caffein) Phenacetin -> Acetaminophen CYP2D6 Dextromethorphan -> Dextrorphan CYP2C9 (ibuprofen) Diclofenac-> 4OH-Diclofenac CYP2E1 (ethanol) Chlorzoxazone -> 6OH-Chlorzoxazone CYP3A4 Testosteron -> 6ßOH-Testosteron Basale activity Induced activity Basale activity Induced activity Basale activity Induced activity The BiomimesysTM Liver provides a suitable model for hiPSC differentiation into liver organoids that displayed mature features at the gene expression and functional level.A 384 well format is being optimized for high throughput phenotypic screening. Toward liver organoids 20 µm 20 µm20 µm 20 µm20 µm DAPI Albumin Desmin Albumin Desmin Albumin Desmin DAPI - Amiodarone + Amiodarone Functional test - Amiodarone-induced lipid accumulation in liver organoids

×