MICROLIVERMATURATION · Gene network-based maturation of embryonic stem cell-derived hepatocytes in a microfabricated array
FP7 — People (Marie Curie Actions)
- Duration
- 2011-03-01 → 2015-02-28
- EU contribution
- €100,000
- Participants
- 1
- Scheme
- MC-IRG
Lines connect the coordinator with its partners.
Results in brief
Gene network-based maturation of embryonic stem cell-derived hepatocytes in a microfabricated array
Embryonic and genetically-induced pluripotent stem cells (hPSCs) have a limitless capacity for self-renewal and a remarkable ability to differentiate, promising a limitless source of cells for cell therapies and tissue engineering. Organ scarcity and lack of proliferative ability make hPSC-derived hepatocytes one of the few potential sources of cells for clinical applications and pharmaceutical drug discovery. However, current techniques for hepatocyte differentiation result in mixed cell populations, minimal metabolic function, and long-term de-differentiation. The aim of this project is to develop and optimize a protocol for the differentiation of hPSCs to hepatocytes following developmental changes in the hepatic transcription factor network. A fundamental discovery of this project was the role of nutrients in modulating key decision points in stem cell differentiation. Our work showed that enhanced glycolysis in hPSC drives the production of Ac-CoA and subsequent histone acetylation. Mitochondrial activation in the first hours of differentiation shuts down this pathway, driving de-acetylation and stem cell commitment (Moussaieff et al. Cell Metabolism 2015). Our second discovery was the role of gut microbiome in the production of key metabolites controlling late stage hepatocyte maturation. We show that the post-partum transition from placental to enteral nutrition exposes the fetal liver to new microbiome-derived metabolites that induce hepatocyte to acquire a mature phenotype. We showed that microbiome-derived lithocholic acid and vitamin K2, act synergistically to induce PXR and activate nascent drug metabolism in hPSC-derived hepatocytes (Avior et al. Hepatology 2015). Our work was the first to demonstrate fully functional hPSC-derived hepatocytes for drug toxicity screening, demonstrate drug toxicity of 12 compounds with accuracy above 95% and R2 of 0.90. 1. Y. Avior, G. Levy, M. Zimerman, D. Kitsberg, R. Schwartz, R. Sadeh, A. Moussaieff, M. Cohen, J. Itskovitz-Eldor, Y. Nahmias. Microbial-Derived Lithocholic Acid and Vitamin K2 Drive the Metabolic Maturation of Pluripotent Stem Cells-Derived and Fetal Hepatocytes, Hepatology epub (2015) 2. A. Moussaieff, M. Rouleau, D. Kitsberg, S.S. Shen-Orr, I.Laevsky, M. Amit, D. Bomze, T. Scherf, M. Nissim-Rafinia, E. Meshorer, J. Itskovitz-Eldor, D. Aberdam, Y. Nahmias. Glycolysis-mediated Changes in Acetyl-coA and Histone Acetylation Control the Early Differentiation of Embryonic Stem Cells, Cell Metabolism 21(3): 392 - 402 (2015) 3. P. Roelandt, S. Obeid, J. Paeshuyse, J. Vanhove, A. Van Lommel, Y. Nahmias, F. Nevens, J. Neyts, C.M. Verfaillie. Human pluripotent stem cell derived hepatocytes support complete replication of hepatitis C virus. J Hepatol. 57(2):246-51 (2012) 4. E. Keinan, E. Ezra, Y. Nahmias. Frame Rate Free Image Velocimetry for Microfluidic Devices, Appl. Phys. Lett. 103, 063507 (2013)
Data: CORDIS, © European Union
Project objective
Embryonic stem cells (ESC) have a limitless capacity for self-renewal a remarkable ability to differentiate, promising a limitless source of cells for cell therapies and tissue engineering. Organ scarcity and lack of proliferative ability makes ESC-derived hepatocytes one of the few potential sources of cells for clinical applications and pharmaceutical drug discovery. However, current techniques for hepatocyte differentiation result in mixed cell populations, minimal metabolic function, and long-term de-differentiation. These differences between mature and differentiated hepatocytes are due to insufficient development of the hepatic transcriptional network during differentiation. Preliminary evidence suggests that modifying the nutritional state of ESC-derived hepatocytes by lowering glucose and increasing fatty acids, mimicking suckling, results in a more mature hepatic phenotype, possibly through the induction of the fatty acid-responsive nuclear receptors, PPARa and HNF4a. This result is interesting as the role of nutrients in differentiation has yet to be elucidated in spite of well known nutritional changes during the pre- and post-natal periods. The hepatic nuclear receptor network is especially important in this context as these ligand-activated transcription factors respond to glucose, sterols, fatty and bile acids offering an epigenetic point of regulation in the development of liver metabolism. The technology which allows us to elucidate and control transcriptional events during differentiation is a microdevice termed the Living Cell Array (LCA). The LCA is a microfabricated device containing multiple GFP reporter cells under microfluidic control allowing high-throughput screening of changes in transcriptional activity during ESC differentiation. The ability of the LCA to monitor transcriptional activity dynamics in high-throughput makes it a unique tool for the study of hepatocyte differentiation under varying nutritional stimulation.
Original text from CORDIS.
Participants
- THE HEBREW UNIVERSITY OF JERUSALEM · JerusalemCoordinatorIsrael
Links
Data: CORDIS, © European Union