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Review
. 2023 Nov 2;24(21):15921.
doi: 10.3390/ijms242115921.

Modeling Liver Development and Disease in a Dish

Waqas Iqbal  1   2   3 , Yaru Wang  1   2   3 , Pingnan Sun  1   2   3 , Xiaoling Zhou  1   2   3
Affiliations
Review

Modeling Liver Development and Disease in a Dish

Waqas Iqbal et al. Int J Mol Sci. .

Abstract

Historically, biological research has relied primarily on animal models. While this led to the understanding of numerous human biological processes, inherent species-specific differences make it difficult to answer certain liver-related developmental and disease-specific questions. The advent of 3D organoid models that are either derived from pluripotent stem cells or generated from healthy or diseased tissue-derived stem cells have made it possible to recapitulate the biological aspects of human organs. Organoid technology has been instrumental in understanding the disease mechanism and complements animal models. This review underscores the advances in organoid technology and specifically how liver organoids are used to better understand human-specific biological processes in development and disease. We also discuss advances made in the application of organoid models in drug screening and personalized medicine.

Keywords: human pluripotent stem cell; liver development; metabolic liver disease; organ-on-chip; organoids; primary liver cancer; regenerative medicine; viral infection.

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Conflict of interest statement

Authors declare no competing interest. The funding body played no role in the design, data collection, interpretation, the analysis of the study, and the writing of the manuscript.

Figures

Figure 4
Figure 4
Organoids in translational medicine. (a) Drug-Screening; Patient-derived organoids can be used in a high-throughput drug screening platform to study DILI. These pre-clinical platforms can be used as diagnostic tools and predictive models for drug discovery [177]. (b) Organ-on-chip; Depiction of a mechanically actuable two channel organ chip fabricated using soft lithography from PDMS with two channels parallel to each other separated by a microporous membrane. Cells from different tissues are cultured on top and bottom of the extracellular matrix (ECM)-coated microporous membrane to emulate tissue–tissue interaction where air introduction above the epithelium can take place to recreate air-liquid interface (as observed in lungs) or the channel could be used for fluid perfusion [169]. Human liver organoid-based chips have recently been used for DILI risk assessment [177]. DILI, drug induced liver injury; PDMS, polydimethylsiloxane.
Figure 1
Figure 1
Timeline of important events in liver organoid technology. Rat liver cells successfully formed spheroidal aggregates in non-adherent plastic substratum [5]. Hepatocytes and fibroblasts co-culture formed 3D structures [6]. Biodegradable hydrogels used to generate liver organoids in vitro [7]. Liver parenchymal and non-parenchymal cells isolated and co-cultured to form organoids [8]. 3D Cell and biomaterial complex formation [9]. Development of bioreactor for the generation of liver organoids using bio-artificial liver [10]. Generation of vascularized liver organoids [11]. Generation of organoids from patient-derived cells [12]. Generation of liver ductal organoids to recapitulate SAR-CoV-2 infection [13]. Development of fetal liver organoids for gene editing using CRISPR-Cas9 [14].
Figure 2
Figure 2
Organogenesis and generation of liver organoids from various sources. (a) Schematic depiction of stages involved in organogenesis. Embryo formation is followed by blastocyst stage. The blastocyst consists of an outer layer of trophectodermal cells and ICM. The ICM, consisting of embryonic stem cells, further specializes into either epiblast lineage or primitive endoderm lineage during the late stages of its formation. Blastocyst is followed by gastrulation, where morphological rearrangements transform epiblast into the three germ layers: ectoderm, endoderm, and mesoderm [16,17]. The endoderm becomes patterned into anterior foregut (AF), posterior foregut (PF), midgut (M), and hindgut (H). As illustrated above, the liver is derived from PF domain of the endoderm. (b) Organoids derived from pluripotent stem cells follow a stage-wise differentiation process that recapitulates signaling pathways observed during development. The differentiation process starts by directing iPSCs/ESCs towards endodermal fate when exposed to Act A and Wnt. The cells are embedded in ECM and differentiated into hepatoblasts-like cells (progenitor cells) using FGF and BMP. Hepatoblast-like cells differentiate into hepatocyte-like cells via exposure to OSM [18,19]. Moreover, ductal organoids can be generated by modulating FGF, EGF, and Act A signaling [16]. Hepatoblasts embedded in ECM give rise to hepatic organoids. Several tissue sources have been used to generate patient-derived organoids using a number of different techniques for tissue processing. The variation in techniques used has led to non-standardized patient-derived organoid culture techniques [20]. In general, tissue-derived stem cells are dissociated into single cells and embedded in extracellular matrix to generate organoids [21,22]. (c) Generation of organoids. Organoids generated for various organs derived from the three germ layers. ICM, inner cell mass; Act A, Activin A; BMP, bone morphogenetic protein; FGF, fibroblast growth factor; ECM, extracellular matrix; HGF, hepatocyte growth factor; OSM, Oncostatin M; Dex, dexamethasone; iPSCs, induced pluripotent stem cells; ESCs, embryonic stem cells; hPSCs, human pluripotent stem cells; ASCs, adult stem cells.
Figure 3
Figure 3
Application of liver organoid models. (a) Virus host interaction. HBV; Modeling HBV infection and drug screening. The model developed can successfully recapitulate HBV infection [21]. Depiction of HBV life cycle from attachment to secretion of viral particles. HBV enters hepatocyte via a mechanism involving NTCP receptor. Once internalized, the HBV’s rcDNA genome liberated into the nucleus is converted into cccDNA, which serves as a template for transcription. The dslDNA produced can either integrate into cellular genome or convert into cccDNA. The viral mRNA transported to cytoplasm is translated into viral proteins. The pgRNA and viral polymerase are encapsulated and reverse transcribed into progeny rcDNA within the nucleocapsid. HCV; The importance of new models to study HCV is crucial due to limited animal models [52]. Organoids derived from adult stem cells and hPSCs can be used to further our understanding of HCV infection and develop antiviral drugs. HCV interaction with cell surface receptors initiates viral entry. HEV; Generation of liver-derived organoids support HEV infection and life cycle and could help develop new therapies. SARS-CoV-2; The interaction of spike protein with ACE2 receptor in the presence of TMPRSS2 facilitates the entry of SARS-CoV-2 into the host cell. COVID-19 caused by SARS-CoV-2 could mediate cell damage, dysregulate RAAS leading to decreased cleavage of angiotensin I and angiotensin II, thromboinflammation, and endothelial cell damage, and inhibit interferon signaling, i.e., depletion of T lymphocytes and production of cytokines such as IL-6 and TNFα [76]. (b) Modeling steatohepatitis in vitro. The failure of animal models in identifying translatable therapies highlights the need for improved models. Generation of organoids from patient-derived hPSCs consisting of multiple hepatic cell types successfully emulates liver-in-a-dish and can be utilized to study liver inflammation and fibrosis and identify effective drug treatments [18]. NTCP, Sodium taurocholate cotransporting polypeptide; rcDNA, relaxed circular DNA; cccDNA, closed circular DNA; dslDNA, double-stranded linear DNA; pgRNA, pregenomic RNA; ACE2, Angiotensin-converting enzyme 2; TMPRSS2, transmembrane protease, serine 2; RAAS, renin-angiotensin-aldosterone system; IL-6, interleukin 6; TNFα, tumor necrosis factor alpha.
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