VEGFA mRNA-LNP promotes biliary epithelial cell-to-hepatocyte conversion in acute and chronic liver diseases and reverses steatosis and fibrosis
Posted on: 4 August 2023
Preprint posted on 18 April 2023
Article now published in Cell Stem Cell at http://dx.doi.org/10.1016/j.stem.2023.10.008
Steer clear of steatosis – VEGFA lipid nanoparticles (LNPs) drive BEC-hepatocyte conversion, an alternative pathway for liver regeneration.
Selected by Organogenesis and Cancer ProgramCategories: developmental biology, molecular biology
Background
In humans, the liver is capable of regenerating following injury primarily through the proliferation of mature hepatocytes. In the context of end-stage liver disease (ESLD) in steatosis, cirrhosis or liver failure, the regenerative ability of hepatocytes is impaired, and injury becomes irreversible. As a result, many researchers have postulated alternative routes of regeneration that could be harnessed to rescue liver function in the context of disease.
Apart from hepatocyte self-renewal, an alternative route of hepatocyte production is hypothesised to originate from biliary epithelial cell (BECs) (Rodrigo-Torres et al. 2014). Often referred to as liver progenitor cells (LPCs), BEC precursors are well-documented to differentiate into de novo hepatocytes during liver disease. In these scenarios, this mainly occurs in highly proliferative and reactive regions of the bile duct, in a process termed ductular reaction (DR). This process has been characterised by the presence hepatocyte-like cells that have been described as bi-phenotypic cells, or “ductular hepatocytes”, that express both BEC and mature hepatocyte markers (Haque et al., 1996; Yoon et al., 2011; Deng et al., 2018). Furthermore, lineage-tracing experiments performed in mice and zebrafish have demonstrated this de novo hepatocyte formation from BECs in the absence of self-renewing hepatocytes (Lu et al, 2015; Raven et al., 2017; Russell., 2018). Despite this, there are currently no therapeutic options that can promote this response in ESLD.
In this preprint, Rizvi and colleagues explored the therapeutic benefits of utilising nucleoside-modified mRNA encapsulated lipid-nanoparticles (mRNA-LNPs) to treat acute and chronic liver injury models. Here, mRNA-LNPs delivering vascular endothelial growth factor A (VEGFA) were used to study its effect on BEC-to-hepatocyte conversion and hallmarks of disease, such as steatosis and fibrosis, in the liver.
Key Findings
VEGFR signalling regulates BEC-to-hepatocyte conversion during liver regeneration in zebrafish and mice
In this study, the authors utilised zebrafish and mice models to study the mechanism of BEC-to-hepatocyte conversion during liver regeneration. Firstly, the authors genetically ablated hepatocytes with the nitroreductase-metronidazole model and found that inhibition of VEGFR signalling, both pharmaceutically and genetically, led to a significant reduction in the number of hepatocytes found in BEC-positive areas during a critical regeneration window. In support of this, overexpressing human VEGFA specifically in BECs during liver injury was also sufficient to promote hepatocyte differentiation in these regions. Together these findings suggest a mechanism for VEGFR signalling in BEC-to-hepatocyte conversion during liver regeneration in zebrafish.
Next, using a diet-induced liver injury mouse model of non-alcoholic steatohepatitis (NASH), the authors tested the impact of VEGFA activation during liver regeneration. In an elegant experiment, chronically liver-injured mice were subjected to AAV-mediated delivery of p21 (to induce hepatocyte senescence), followed by two injections of mRNA-LNPs containing VEGFA. In mice that received VEGFA mRNA-LNP injections, cross-sections and flow cytometry analysis revealed a greater proportion of newly developed hepatocytes in these livers, compared to livers of mice injected with control LNPs. In addition to this, new hepatocytes in VEGFA mRNA-LNP injected mice were found to be adjacent to KRT7-positive BECs, indicative of their BEC origin. Interestingly, VEGFA mRNA-LNP treatment significantly reduced fibrosis and steatosis in the liver, and in addition, new hepatocytes were found to be as sufficient at storing glycogen when compared to neighbouring pre-existing hepatocytes. VEGFA mRNA-LNP-treated livers were also functionally more efficient at clearing lipids from hepatocytes compared to control LNP livers. Together these findings demonstrate a potential clinical benefit of VEGFA mRNA-LNPs in alleviating symptoms inflicted by NASH.
VEGFA promotes BEC-to-hepatocyte conversion in APAP-induced acute liver injury in mice
To model human acute liver injury, acute acetaminophen (Paracetomol) toxicity (APAP) was induced in mice. Strikingly, after two consecutive injections of VEGFA mRNA-LNPs, the livers of these mice showed a 20-fold increase in the proportion of newly developed hepatocytes, compared to the livers of mice injected with control LNPs. As well as being situated close to bile ducts, these new BEC-derived hepatocytes were also found to be as sufficient at storing glycogen as the pre-existing hepatocytes. Together with the chronic liver injury model, these results demonstrate the potential clinical benefit of VEGFA mRNA-LNPs as a tool to promote liver regeneration.
VEGFA-KDR signalling may have a conserved role in BEC-derived hepatocyte formation
The authors next investigated KDR, the receptor for VEGFA, in liver regeneration in the context of both chronic diet-induced injury and acute APAP-induced injury. The identification of a BEC subset that express the VEGFA receptor, KDR, led to the hypothesis that that these cells may be precursors of BEC-derived hepatocytes. Using a lineage tracing tool, the authors reported that transient KDR expression in BECs correlates with improved BEC-driven hepatocyte repopulation following VEGFA mRNA-LNP treatment in both injury models. Furthermore, in the acute APAP-injury model, identification of cells co-expressing hepatocyte markers and a reporter for KDR expression supported this hypothesis. In mice, these results demonstrate that VEGFA overexpression in conjunction with BEC-KDR expression during liver injury allow for improved hepatocyte recovery.
Finally, the authors asked whether VEGFA-KDR signalling has a conserved role in BEC-hepatocyte conversion using samples from end-stage liver disease (NASH or alcoholic cirrhosis) and terminal liver failure (Child-Pugh B and C cirrhosis). From these samples, the authors identified bi-phenotypic BECs at the fibrotic septa that expressed not only BEC markers but also hepatocyte markers and vice versa. Importantly, KDR expression was indicated in BECs adjacent to newly generated hepatocytes. This data is an important proof of concept for the hypothesis that KDR-expressing BECs may give rise to intermediate hepatocytes, which represents a novel therapeutic avenue for preventing and treating liver disease.
What I like about this preprint
The elephant in the room is the lipid-nanoparticle (LNP) delivery system. Over recent years it has been gaining attention as a method of mRNA delivery for COVID vaccines and now it is more and more being used in experimental protocols. This preprint is the second study I’ve seen using LNPs to deliver a growth factor (Szöke D, Kovács G, Kemecsei É, et al Nat Commun 2021) and this work is exploring a tool which has an extraordinary range of applications – from animal models to therapeutic delivery.
This preprint combines both zebrafish and mice models with human tissue samples to address a simple question about the VEGFA-KDR pathway in BEC-hepatocyte conversion. Both pharmacological and genetic experiments in zebrafish as well as lineage tracing in clinically-relevant mice models of acute and chronic liver injury allowed the authors to infer the delineating presence of KDR-expressing BECs to KDR-expressing hepatocytes.
Questions for the Author(s)
- Have the authors looked at the local vasculature response to VEGFA-LNP treatment either in healthy or injured mice livers and whether this could be contributing to hepatocyte repopulation?
- Do you believe that the results could be replicated in injury models not specific to hepatocytes in mice (I.e. liver resection or cryoinjury) and would it be beneficial to enhance BEC-hepatocyte conversion in the presence of healthy hepatocytes with the capacity to regenerate?
References:
Rodrigo-Torres, D., Affo, S., Coll, M., Morales-Ibanez, O., Millan, C., Blaya, D., Alvarez-Guaita, A., Rentero, C., Lozano, J.J., Maestro, M.A., et al. (2014). The biliary epithelium gives rise to liver progenitor cells. Hepatology 60, 1367-1377.
Haque, S., Haruna, Y., Saito, K., Nalesnik, M.A., Atillasoy, E., Thung, S.N., and Gerber, M.A. (1996). Identification of bipotential progenitor cells in human liver regeneration. Lab Invest 75, 699-705.
Yoon, S.M., Gerasimidou, D., Kuwahara, R., Hytiroglou, P., Yoo, J.E., Park, Y.N., and Theise, N.D. (2011). Epithelial cell adhesion molecule (EpCAM) marks hepatocytes newly derived from stem/progenitor cells in humans. Hepatology 53, 964-973.
Deng, X., Zhang, X., Li, W., Feng, R.X., Li, L., Yi, G.R., Zhang, X.N., Yin, C., Yu, H.Y., Zhang, J.P., et al. (2018). Chronic Liver Injury Induces Conversion of Biliary Epithelial Cells into Hepatocytes. Cell Stem Cell 23, 114-122.
Lu, W.Y., Bird, T.G., Boulter, L., Tsuchiya, A., Cole, A.M., Hay, T., Guest, R.V., Wojtacha, D., Man, T.Y., Mackinnon, A., et al. (2015). Hepatic progenitor cells of biliary origin with liver repopulation capacity. Nat Cell Biol 17, 971-983.
Raven, A., Lu, W.Y., Man, T.Y., Ferreira-Gonzalez, S., O’Duibhir, E., Dwyer, B.J., Thomson, J.P., Meehan, R.R., Bogorad, R., Koteliansky, V., et al. (2017). Cholangiocytes act as facultative liver stem cells during impaired hepatocyte regeneration. Nature 547, 350-354.
Russell, J.O., Lu, W.Y., Okabe, H., Abrams, M., Oertel, M., Poddar, M., Singh, S., Forbes, S.J., and Monga, S.P. (2018). Hepatocyte-specific beta-catenin deletion during severe liver injury provokes cholangiocytes to differentiate into hepatocytes. Hepatology. 10.1002/hep.30270.
Szőke, D., Kovács, G., Kemecsei, É., Bálint, L., Szoták-Ajtay, K., Aradi, P., Styevkóné Dinnyés, A., Mui, B.L., Tam, Y.K., Madden, T.D., Karikó, K., Kataru, R.P., Hope, M.J., Weissman, D., Mehrara, B.J., Pardi, N., and Jakus, Z. (2021). Nucleoside-modified VEGFC mRNA induces organ-specific lymphatic growth and reverses experimental lymphedema. Nat Commun 12, 3460
doi: https://doi.org/10.1242/prelights.35254
Read preprintSign up to customise the site to your preferences and to receive alerts
Register hereAlso in the developmental biology category:
Germplasm stability in zebrafish requires maternal Tdrd6a and Tdrd6c
Justin Gutkowski
Cellular signalling protrusions enable dynamic distant contacts in spinal cord neurogenesis
Ankita Walvekar
Actin-based deformations of the nucleus control multiciliated ependymal cell differentiation
Ryan Harrison
Also in the molecular biology category:
Germplasm stability in zebrafish requires maternal Tdrd6a and Tdrd6c
Justin Gutkowski
Platelet-derived LPA16:0 inhibits adult neurogenesis and stress resilience in anxiety disorder
Harvey Roweth
Green synthesized silver nanoparticles from Moringa: Potential for preventative treatment of SARS-CoV-2 contaminated water
Safieh Shah, Benjamin Dominik Maier
preListsdevelopmental biology category:
in theBSDB/GenSoc Spring Meeting 2024
A list of preprints highlighted at the British Society for Developmental Biology and Genetics Society joint Spring meeting 2024 at Warwick, UK.
List by | Joyce Yu, Katherine Brown |
GfE/ DSDB meeting 2024
This preList highlights the preprints discussed at the 2024 joint German and Dutch developmental biology societies meeting that took place in March 2024 in Osnabrück, Germany.
List by | Joyce Yu |
‘In preprints’ from Development 2022-2023
A list of the preprints featured in Development's 'In preprints' articles between 2022-2023
List by | Alex Eve, Katherine Brown |
preLights peer support – preprints of interest
This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.
List by | preLights peer support |
The Society for Developmental Biology 82nd Annual Meeting
This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.
List by | Joyce Yu, Katherine Brown |
CSHL 87th Symposium: Stem Cells
Preprints mentioned by speakers at the #CSHLsymp23
List by | Alex Eve |
Journal of Cell Science meeting ‘Imaging Cell Dynamics’
This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.
List by | Helen Zenner |
9th International Symposium on the Biology of Vertebrate Sex Determination
This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.
List by | Martin Estermann |
Alumni picks – preLights 5th Birthday
This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.
List by | Sergio Menchero et al. |
CellBio 2022 – An ASCB/EMBO Meeting
This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.
List by | Nadja Hümpfer et al. |
2nd Conference of the Visegrád Group Society for Developmental Biology
Preprints from the 2nd Conference of the Visegrád Group Society for Developmental Biology (2-5 September, 2021, Szeged, Hungary)
List by | Nándor Lipták |
Fibroblasts
The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!
List by | Osvaldo Contreras |
EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)
A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.
List by | Alex Eve |
EMBL Conference: From functional genomics to systems biology
Preprints presented at the virtual EMBL conference "from functional genomics and systems biology", 16-19 November 2020
List by | Jesus Victorino |
Single Cell Biology 2020
A list of preprints mentioned at the Wellcome Genome Campus Single Cell Biology 2020 meeting.
List by | Alex Eve |
Society for Developmental Biology 79th Annual Meeting
Preprints at SDB 2020
List by | Irepan Salvador-Martinez, Martin Estermann |
FENS 2020
A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020
List by | Ana Dorrego-Rivas |
Planar Cell Polarity – PCP
This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.
List by | Ana Dorrego-Rivas |
Cell Polarity
Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.
List by | Yamini Ravichandran |
TAGC 2020
Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20
List by | Maiko Kitaoka et al. |
3D Gastruloids
A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.
List by | Paul Gerald L. Sanchez and Stefano Vianello |
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
List by | Madhuja Samaddar et al. |
EDBC Alicante 2019
Preprints presented at the European Developmental Biology Congress (EDBC) in Alicante, October 23-26 2019.
List by | Sergio Menchero et al. |
EMBL Seeing is Believing – Imaging the Molecular Processes of Life
Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019
List by | Dey Lab |
SDB 78th Annual Meeting 2019
A curation of the preprints presented at the SDB meeting in Boston, July 26-30 2019. The preList will be updated throughout the duration of the meeting.
List by | Alex Eve |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
List by | Rob Hynds |
Young Embryologist Network Conference 2019
Preprints presented at the Young Embryologist Network 2019 conference, 13 May, The Francis Crick Institute, London
List by | Alex Eve |
Pattern formation during development
The aim of this preList is to integrate results about the mechanisms that govern patterning during development, from genes implicated in the processes to theoritical models of pattern formation in nature.
List by | Alexa Sadier |
BSCB/BSDB Annual Meeting 2019
Preprints presented at the BSCB/BSDB Annual Meeting 2019
List by | Dey Lab |
Zebrafish immunology
A compilation of cutting-edge research that uses the zebrafish as a model system to elucidate novel immunological mechanisms in health and disease.
List by | Shikha Nayar |
Also in the molecular biology category:
2024 Hypothalamus GRC
This 2024 Hypothalamus GRC (Gordon Research Conference) preList offers an overview of cutting-edge research focused on the hypothalamus, a critical brain region involved in regulating homeostasis, behavior, and neuroendocrine functions. The studies included cover a range of topics, including neural circuits, molecular mechanisms, and the role of the hypothalamus in health and disease. This collection highlights some of the latest advances in understanding hypothalamic function, with potential implications for treating disorders such as obesity, stress, and metabolic diseases.
List by | Nathalie Krauth |
BSCB-Biochemical Society 2024 Cell Migration meeting
This preList features preprints that were discussed and presented during the BSCB-Biochemical Society 2024 Cell Migration meeting in Birmingham, UK in April 2024. Kindly put together by Sara Morais da Silva, Reviews Editor at Journal of Cell Science.
List by | Reinier Prosee |
‘In preprints’ from Development 2022-2023
A list of the preprints featured in Development's 'In preprints' articles between 2022-2023
List by | Alex Eve, Katherine Brown |
CSHL 87th Symposium: Stem Cells
Preprints mentioned by speakers at the #CSHLsymp23
List by | Alex Eve |
9th International Symposium on the Biology of Vertebrate Sex Determination
This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.
List by | Martin Estermann |
Alumni picks – preLights 5th Birthday
This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.
List by | Sergio Menchero et al. |
CellBio 2022 – An ASCB/EMBO Meeting
This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.
List by | Nadja Hümpfer et al. |
EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)
A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.
List by | Alex Eve |
FENS 2020
A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020
List by | Ana Dorrego-Rivas |
ECFG15 – Fungal biology
Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome
List by | Hiral Shah |
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
List by | Madhuja Samaddar et al. |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
List by | Rob Hynds |
MitoList
This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.
List by | Sandra Franco Iborra |