iLite - innovation in Liver tissue engineering (2016-2022)
iLite (2016 - 2021)
The goals of this project are to build an external bioartificial liver, a liver-on-chip as well as a bioengineered liver. In the present project, bioconstruction of the liver will be performed by the assembly of building blocks –liver organoids, vascular networks and biliary networks- built separately and then integrated into a scaffold.
Funding
iLite (2016 - 2021)
The goals of this project are to build an external bioartificial liver, a liver-on-chip as well as a bioengineered liver.
In the present project, bioconstruction of the liver will be performed by the assembly of building blocks –liver organoids, vascular networks and biliary networks- built separately and then integrated into a scaffold. Vascular and biliary networks will be produced by a mix of technologies comprising organotypic culture, micropatterning and stereolithography.
Liver organoids will be produced by bioprinting as sheets or spheroids, involvinghepatocytes, endothelial cells and cholangiocytes, or as liver buds obtained from stem cells.We will focus particularly on producing a functional biliary network which has been a missing element in all previous studies. The integration step should lead to a functional complete vascularized transplantable liver with a bile duct.
The different steps of bioconstruction will be modeled in silico. Function of the different constructs will be assessed in vitro by functional assays and in vivo after implantation in animal models, which will also favor a complete maturation of the construct. Taking into account the poor yield of mature human liver cells, most cells will be obtained from differentiation of pluripotent stem cells, either embryonic or induced, which are now well mastered by our teams. Final assessment of the liver will be performed in an immunodeficient rat model of acute liver insufficiency.
Progress in the construction of functional liver organoids will be transposed to the construction of an external bioartificial liver for the temporary treatment of acute liver failure and in liver- on-chip microfluidic devices for the study of drug metabolism and predictive toxicology. Studies in microfluidic devices should also allow us to improve maturation of the liver bioconstruction thanks to fluid pressure and flow. By the end of the 5-year project, we aim to provide a prototype of an external bioartificial liver to be used for phase I and II clinical studies as well as a prototype liver-on-chip device ready for use by pharmaceutical companies producing new drugs. All steps in the project will take into consideration GMP procedures.
These goals can be reached by an interdisciplinary approach bringing together researcher from several Universities, Research Institute and small companies who master innovative technologies of bioconstruction.
WP1 - Imaging, image analysis, in silico models, and software generation
- Imaging techniques and protocols for liver tissue sub-types
- Image processing and analysis of tissue and cell morphologies from normal and artificial liver
- Mathematical models of the hepatocyte and cholangiocyte gene regulatory networks
- Systems biology computational models of functional unit components and of corresponding fluids and metabolites fate
WP2 - Liver tissue/organoids
- Sheets of hepatocytes/endothelial cells
- Pluricellular spheroids, organization and functionality
- Spontaneously organized or 3D micropatterned/bioprinted pluricellular organoids, organization and functionality
- Functional organoids in a mouse model of acute liver failure
WP3 - Construction of a biliary duct network
- 2D and 3D biliary networks obtained by micropatterning and bioprinting
WP4 - In vitro construction of a vascular network
- 2D and 3D vascular networks
- Liver-on-chips including a functional vascular network
WP5 - Integration of liver organoids into vascular and/or biliary networks and scale-up
- Liver organoids with a connected vascular network
- Liver organoids with a connected vascularized biliary network
- Integration of both of the above towards an implantable liver substitute
WP6 - Liver-on-chip
- Devices for drug screening and/or predictive toxicology with endothelium/hepatocyte co-culture
- Microfluidic devices as surrogates for canal of Hering and sinusoid networks
- Microfluidic devices with bio-printed liver organoids
WP7 - External bio-artificial liver
- EBAL prototype, validated by preclinical studies and ready for industrialization
- Bio-hydrid tissues (cells and materials) to be inserted in EBAL
- Treatment protocol to be challenged in further clinical studies
WP8 - Safety / GMP / Industrialization
- Single core elements for automated, standardized, traceable, cost-effective, safe and regulatory- compliant manufacturing of ATMPs: master bank of cGMP hESC-derived hepatocytes, large-scale production of hESC-derived hepatocytes in a closed system
- Transfer 3D bioprinting technologies, products and knowhow to existing or de novo startups
- Industrialization of anEBAL
- Industrialization of a liver-on-chip device
Publications
Publications under the iLite project (ANR-16-RHUS-0005), the aid of which is financed by the “Investissements d’Avenir” program (Investment Program for the Future) launched by the State and implemented by the ANR (National Research Agency)
A versatile microfluidic tool for the 3D culture of HepaRG cells seeded at various stages of differentiation
Advanced techniques and awaited clinical applications for human pluripotent stem cell differentiation into hepatocytes
Analysis of hiPSCs differentiation toward hepatocyte-like cells upon extended exposition to oncostatin.
Analysis of the behavior of 2D monolayers and 3D spheroid human pancreatic beta cells derived from induced pluripotent stem cells in a microfluidic environment.
Analysis of the transcription factors and their regulatory roles during a step-by-step differentiation of induced pluripotent stem cells into hepatocyte-like cells.
Bioengineering Organs for Blood Detoxification.
Biophysical Control of Bile Duct Epithelial Morphogenesis in Natural and Synthetic Scaffold.
Channeled polysaccharide-based hydrogel reveals influence of curvature to guide endothelial cell arrangement in vessel-like structures.
Characterisation of the liver zonation like transcriptomic patterns in HLCs derived from hiPCs in a microfluidic biochip environment
Characterization of the proteome and metabolome of human liver sinusoidal endothelial-like cells derived from induced pluripotent stem cells
Construction of functional biliary epithelial branched networks with predefined geometry using digital light stereolithography
Convergence of microengineering and cellular self-organization towards functional tissue manufacturing.
Cryogel-Integrated Biochip for Liver Tissue Engineering
Development of a pancreas-liver organ-on-chip coculture model for organ-to-organ interaction studies.
Development of 3D Hepatic Constructs Within Polysaccharide-Based Scaffolds withTunable Properties.
Generation and Quantitative Characterization of Functional and Polarized Biliary Epithelial Cysts.
HepaRG Self-Assembled Spheroids in Alginate Beads Meet the Clinical Needs for Bioartificial Liver.
Influence of cell mechanics in embryonic bile duct lumen formation: insight from quantitative modeling.
Influence of CPM-dependent sorting on the multi-omics profile of hepatocyte-like cells matured in microscale biochips
Integration of an oxygen sensor into a polydymethylsiloxane hepatic culture T device for two-dimensional gradient characterization.
Integration of metabolomic and transcriptomic profiles of hiPSCs-derived hepatocytes in a microfluidic environment.
Integration of metabolomic and transcriptomic profiling to compare two protocols of differentiation of human induced pluripotent stem cells into hepatocyte.
Investigation of the hepatic respiration and liver zonation on rat hepatocytes using an integrated oxygen biosensor in a microscale device.
Quantitative agent- based modeling reveals mechanical stress response of growing tumor spheroids is predictable over various growth conditions and cell lines.
Quantifying the mechanics and growth of cells and tissues in 3D using high resolution computational models.
Microwell-based pancreas-on-chip model enhances genes expression and functionality of rat islets of Langerhans.
Multi-omics analysis of hiPSCs-derived HLCs matured on-chip revealed patterns typical of liver regeneration.
Nanotechnologies et ingénierie du foie bioartificiel. Une autre idée de la « convergence technologique ».
Pluripotent stem cell-derivedcholangiocytes andcholangiocyte organoids.
Pluripotent-Stem-Cell-Derived Hepatic Cells: Hepatocytes and Organoids for Liver Therapy and Regeneration.
Preclinical characterization of alginate‐Poly‐L‐Lysine encapsulated HepaRG for extracorporeal liver supply.
Profiling of derived-hepatocyte progenitors from induced pluripotent stem cells using nanoCAGE promoter analysis.
Simulation of a detoxifying organ function: focus on hemodynamics modeling and convection-reaction numerical simulation in microcirculatory networks.
The endothelium, a key actor in organ development and hPSC-derived organoid vascularization
Transcriptome profiling of hiPSC-derived LSECs with nanoCAGE.