Enhanced Preprints

Novel 3D Approach to Model Non-Alcoholic Fatty Liver Disease using human Pluripotent Stem Cells

  1. Carola Maria Morell
  2. Samantha Grace Tilson
  3. Rute Alexandra Tomaz
  4. Arash Shahsavari
  5. Andi Munteanu
  6. Giovanni Canu
  7. Brandon Tyler Wesley
  8. Marion Perrin
  9. Imbisaat Geti
  10. Subhankar Mukhopadhyay
  11. Francesca Mazzacuva
  12. Paul Gissen
  13. Jose Garcia-Bernardo
  14. Martin Bachman
  15. Casey Allison Rimland
  16. Fotios Sampaziotis
  17. Irina Mohorianu
  18. Ludovic Vallier author has email address
  1. Wellcome-MRC Cambridge Stem Cell Institute, JCBC Cambridge Biomedical Campus, Dept of Surgery, University of Cambridge, Cambridge, UK
  2. Liver Disease Branch, NIDDK, NIH, Bethesda, Maryland, USA
  3. Wellcome Trust Sanger Institute, Hinxton, UK
  4. UCL Great Ormond Street Institute of Child Health, London, London, UK
  5. NIHR Great Ormond Street Hospital Biomedical Research Centre, London, UK
  6. Medicines Discovery Catapult, Alderley Park, UK
  7. Immunopathogenesis Section, Laboratory of Parasitic Diseases, NIAID, NIH, Bethesda, MD
  8. Berlin Institute of Health Centre for Regenerative Therapies, Berlin, Germany
  9. Max Planck Institute for Molecular Genetics, Berlin, Germany

Peer review process

Not revised: This Reviewed Preprint includes the authors’ original preprint (without revision), an eLife assessment, and public reviews.

Read more about eLife’s peer review process.

Editors

  • Reviewing Editor
    Pramod Mistry
    Yale University, New Haven, United States of America
  • Senior Editor
    Pramod Mistry
    Yale University, New Haven, United States of America

Reviewer #1 (Public Review):

There is an undisputable need for better in vitro models recapitulating steatotic liver diseases. This article is from a group of well-known stem cell experts that use human induced pluripotent stem cells (hiPSCs) to build a multicellular steatosis model in vitro. While the model is strong for testing hepatocytes responses, it falls short on translational aspects as well as on non-parenchymal liver cells.

(1) The authors should use the new nomenclature for the disease, MASLD / MASH, as proposed by the scientific societies (Rinella ME, et al. J Hepatol. 2023; 79(6):1542-1556. PMID: 37364790).

(2) There has been a similar approach by the Takebe group (Ouchi R, et al., Cell Metab. 2019; 30(2):374-384, PMID: 31155493). What is different in this model?

(3) The work is very technical and does neither provide any new mechanistic insights nor does it test any new interventions. I do see the clear technical advance in the long-term culture. However, I do not see that this system would allow modelling true "chronic" changes in MASLD, e.g. steatohepatitis and/or fibrosis.

(4) While I am very convinced about the validity of the "hepatocyte" component in this system, the NPC compartment is insufficient. The 3D model does certainly not contain Kupffer cells (which have very distinct characteristics from "M0" macrophages) and does not contain true HSCs (LX-2 is a very insufficient model). Also, the model lacks flow conditions, which does not allow to factor in pathogenic signals from the circulation / portal vein (e.g. gut-liver axis). This will only allow very limited insights into the crosstalk between hepatocytes and NPCs.

(5) The translational value of this model remains unclear to me. The scRNA-seq data should be meticulously compared to sc/snRNA-seq data from human MASLD livers at different stages to understand, what this system is able to model (maybe very early stages of steatosis?).

(6) The study lacks a "use case" to study interventions, e.g. testing resmetirom or any other of the new MASLD drugs in this system.

Reviewer #2 (Public Review):

Summary:

The authors developed a 3D multi-cellular platform mimicking the complex interplays involved in the pathogenesis of NAFLD/NASH by employing hiPSCs-derived parenchymal and non-parenchymal cells in combination of organoids obtained from primary human cholangiocytes and the human hepatic stellate cell line LX2. They show that hiPSC-derived hepatocyte are able to accumulate intracellular lipids in fashion similar to human NAFLD and that prolonged accumulation leads to activation of inflammatory and fibrogenic pathways.

Strengths:

This is an original attempt to create a 3D all-human multicellular cellular platform recapitulating human NAFLD/NASH. The results are very encouraging. It is of particular note the fact that fibrogenic markers in the 3D system are not extremely (artificially) activated as in the classic 2D system. This makes the proposed platform more realistic.

Weaknesses:

The mixture of hiPSC-derived cells and primary or cell-line cells is understandable although potentially adding some variability to the system. The only unclear aspect is the characteristic of the collagen used to create the 3D system. Which type of collagen? Human? Which stiffness?

  1. Howard Hughes Medical Institute
  2. Wellcome Trust
  3. Max-Planck-Gesellschaft
  4. Knut and Alice Wallenberg Foundation