Mini liver organoid models emulating metabolic dysfunction-associated steatotic disease development: gradually emerging beneath the iceberg

Abstract

Tissue spatial interaction in the liver, including the interplay among immune cells, blood vessels, sinusoidal endothelial cells, bile ducts, stellate cells, hepatic progenitor cells, and hepatocytes, is crucial not only for normal organogenesis but also for disease development. Metabolic dysfunction-associated steatotic liver disease (MASLD) has emerged as the most prevalent liver disease, with heterogeneous mechanisms involving cross-talk between hepatocytes and stromal cells under metabolic stress, inflammation, and fibrosis. Developing 3D coculture liver organoid platforms provides novel insights into the complex links of cell communication during disease progression. Thus, we recapitulate liver organoids as models for indicating reconstructed inter-tissue interactions of MASLD, highlighting the potential of transplanting liver organoids as a therapeutic application.

Keywords

MASLD / liver organoid / 3D coculture / disease progression

References

[1] Wentworth BJ.Metabolic dysfunction-associated steatotic liver disease throughout the liver transplant cycle: a comprehensive review.Metab Target Organ Damage2025;5:2

[2] Zheng Q.Epigenetic and epitranscriptomic regulations of metabolic dysfunction-associated steatotic liver disease.Metab Target Organ Damage2024;4:43

[3] Targher G,Tilg H.MASLD: a systemic metabolic disorder with cardiovascular and malignant complications.Gut2024;73:691-702

[4] Stefan N,Neuschwander-Tetri BA.Metabolic dysfunction-associated steatotic liver disease: heterogeneous pathomechanisms and effectiveness of metabolism-based treatment.Lancet Diabetes Endocrinol2025;13:134-48

[5] Ajayi T,Chen S,Oseini A.Surging liver transplantation for nonalcoholic steatohepatitis from 2000-2022: a national database study.South Med J2024;117:302-10 PMCID:PMC11160975

[6] Noureddin M,Bresee C.NASH leading cause of liver transplant in women: updated analysis of indications for liver transplant and ethnic and gender variances.Am J Gastroenterol2018;113:1649-59 PMCID:PMC9083888

[7] Younossi ZM,Ong J.Nonalcoholic steatohepatitis is the most rapidly increasing indication for liver transplantation in the United States.Clin Gastroenterol Hepatol2021;19:580-9.e5

[8] Hu Y,Wang L.Research progress and application of liver organoids for disease modeling and regenerative therapy.J Mol Med2024;102:859-74 PMCID:PMC11213763

[9] Yuan X,Sun Z.Preclinical efficacy and safety of encapsulated proliferating human hepatocyte organoids in treating liver failure.Cell Stem Cell2024;31:484-98.e5

[10] Zhang W,Du Y.Liver cell therapies: cellular sources and grafting strategies.Front Med2023;17:432-57

[11] Lotto J,Hoodless PA.Fetal liver development and implications for liver disease pathogenesis.Nat Rev Gastroenterol Hepatol2023;20:561-81

[12] Nevzorova YA,Cubero FJ.Animal models for liver disease - a practical approach for translational research.J Hepatol2020;73:423-40

[13] Farrell G,Leclercq I.Mouse models of nonalcoholic steatohepatitis: toward optimization of their relevance to human nonalcoholic steatohepatitis.Hepatology2019;69:2241-57

[14] Saxton SH.2D and 3D liver models.J Hepatol2023;78:873-5

[15] Shirini K,Meier RPH.Xenotransplantation literature update July-December 2024.Xenotransplantation2025;32:e70027 PMCID:PMC11856842

[16] Segovia-Miranda F,Kücken M.Three-dimensional spatially resolved geometrical and functional models of human liver tissue reveal new aspects of NAFLD progression.Nat Med2019;25:1885-93 PMCID:PMC6899159

[17] Villanueva MT.Organoids illuminate NAFLD pathogenesis.Nat Rev Drug Discov2023;22:269

[18] Marsee A, Roos FJM, Verstegen MMA, et al; HPB Organoid Consortium. Building consensus on definition and nomenclature of hepatic, pancreatic, and biliary organoids. Cell Stem Cell. 2021;28:816-32. PMCID:PMC11699540

[19] Tang XY,Wang D.Human organoids in basic research and clinical applications.Signal Transduct Target Ther2022;7:168 PMCID:PMC9127490

[20] Cao W,Liu J.LGR5 marks targetable tumor-initiating cells in mouse liver cancer.Nat Commun2020;11:1961 PMCID:PMC7181628

[21] Cao W,Bolkestein M.Dynamics of proliferative and quiescent stem cells in liver homeostasis and injury.Gastroenterology2017;153:1133-47

[22] Lin Y,Liu HJ.HGF/R-spondin1 rescues liver dysfunction through the induction of Lgr5⁺ liver stem cells.Nat Commun2017;8:1175 PMCID:PMC5660090

[23] Huch M,Boj SF.In vitro expansion of single Lgr5⁺ liver stem cells induced by Wnt-driven regeneration.Nature2013;494:247-50 PMCID:PMC3634804

[24] Prior N,Rost F.Lgr5⁺ stem and progenitor cells reside at the apex of a heterogeneous embryonic hepatoblast pool.Development2019;146:dev174557 PMCID:PMC6602348

[25] Schneeberger K,Ye S.Large-scale production of LGR5-positive bipotential human liver stem cells.Hepatology2020;72:257-70 PMCID:PMC7496924

[26] Tonooka Y,Toyoshima M,Kitamoto S.Suppression of the epithelial-mesenchymal transition and maintenance of the liver functions in primary hepatocytes through dispersion culture within a dome-shaped collagen matrix.Biol Pharm Bull2024;47:1241-7

[27] He Y,Zhu X.Large-scale formation and long-term culture of hepatocyte organoids from streamlined in vivo genome-edited GGTA1^-/- pigs for bioartificial liver applications.Xenotransplantation2024;31:e12878

[28] Luo M,Zhang E.Rapid self-assembly mini-livers protect mice against severe hepatectomy-induced liver failure.Adv Sci2024;11:e2309166 PMCID:PMC11151005

[29] Mathapati S,Impellizzeri AA.Small-molecule-directed hepatocyte-like cell differentiation of human pluripotent stem cells.Curr Protoc Stem Cell Biol2016;38:1G.6.1-18

[30] Septiana WL,Antarianto RD.Induced pluripotent stem cells (Ipscs) based liver organoid: the benefits and challenges.Cell Physiol Biochem2023;57:345-59

[31] Hidalgo-Álvarez J,Vera Cruz D,Bianco AC.Localized T3 production modifies the transcriptome and promotes the hepatocyte-like lineage in iPSC-derived hepatic organoids.JCI Insight2023;8:e173780 PMCID:PMC10795825

[32] Reza HA,Reza AA.Synthetic augmentation of bilirubin metabolism in human pluripotent stem cell-derived liver organoids.Stem Cell Reports2023;18:2071-83 PMCID:PMC10679658

[33] Septiana WL,Gunardi H.Liver organoids cocultured on decellularized native liver scaffolds as a bridging therapy improves survival from liver failure in rabbits.In Vitro Cell Dev Biol Anim2023;59:747-63

[34] Tsuzuki S,Okumura T,Ueno Y.PDGF receptors and signaling are required for 3D-structure formation and differentiation of human iPSC-derived hepatic spheroids.Int J Mol Sci2023;24:7075 PMCID:PMC10138705

[35] McCarron S,Conlon DM.Functional characterization of organoids derived from irreversibly damaged liver of patients with NASH.Hepatology2021;74:1825-44

[36] Salas-Silva S,Kim TH.Human chemically-derived hepatic progenitors (hCdHs) as a source of liver organoid generation: application in regenerative medicine, disease modeling, and toxicology testing.Biomaterials2023;303:122360

[37] Hess A,Saad AB.Author correction: single-cell transcriptomics stratifies organoid models of metabolic dysfunction-associated steatotic liver disease.EMBO J2024;43:6792-5 PMCID:PMC11649770

[38] Ietto G,Gritti M.Multicellular liver organoids: generation and importance of diverse specialized cellular components.Cells2023;12:1429 PMCID:PMC10217024

[39] Jo S,Kim H.hiPSC-derived macrophages improve drug sensitivity and selectivity in a macrophage-incorporating organoid culture model.Biofabrication2024;16:035021

[40] Li Y,Yang X.Integration of Kupffer cells into human iPSC-derived liver organoids for modeling liver dysfunction in sepsis.Cell Rep2024;43:113918

[41] Li H,Wang T.Hepatobiliary organoids differentiated from hiPSCs relieve cholestasis-induced liver fibrosis in nonhuman primates.Int J Biol Sci2024;20:1160-79 PMCID:PMC10878143

[42] Wu F,Ren Y.Generation of hepatobiliary organoids from human induced pluripotent stem cells.J Hepatol2019;70:1145-58

[43] Davoodi P,Hassan M,Vosough M.Bioengineering vascularized liver tissue for biomedical research and application.Scand J Gastroenterol2024;59:623-9

[44] Stefan N,Birkenfeld AL,White MF.The role of hepatokines in NAFLD.Cell Metab2023;35:236-52 PMCID:PMC10157895

[45] Correia de Sousa M,Berthou F.Hepatic miR-149-5p upregulation fosters steatosis, inflammation and fibrosis development in mice and in human liver organoids.JHEP Rep2024;6:101126 PMCID:PMC11388170

[46] Kimura M,Iwasawa K.En masse organoid phenotyping informs metabolic-associated genetic susceptibility to NASH.Cell2022;185:4216-32.e16 PMCID:PMC9617783

[47] Gwag T,Li Z.Platelet-derived thrombospondin 1 promotes immune cell liver infiltration and exacerbates diet-induced steatohepatitis.JHEP Rep2024;6:101019 PMCID:PMC10918562

[48] Lee J,Park H.A multicellular liver organoid model for investigating hepatitis C virus infection and nonalcoholic fatty liver disease progression.Hepatology2024;80:186-201

[49] Qi L,Pereira A.Human iPSC-derived proinflammatory macrophages cause insulin resistance in an isogenic white adipose tissue microphysiological system.Small2023;19:e2203725 PMCID:PMC10502939

[50] Qi L,Sharma A.Adipocyte inflammation is the primary driver of hepatic insulin resistance in a human iPSC-based microphysiological system.Nat Commun2024;15:7991 PMCID:PMC11393072

[51] Cherubini A, Ostadreza M, Jamialahmadi O, et al; EPIDEMIC Study Investigators. Interaction between estrogen receptor-α and PNPLA3 p.I148M variant drives fatty liver disease susceptibility in women. Nat Med. 2023;29:2643-55. PMCID:PMC10579099

[52] Hendriks D,Hamer K.Engineered human hepatocyte organoids enable CRISPR-based target discovery and drug screening for steatosis.Nat Biotechnol2023;41:1567-81 PMCID:PMC10635827

[53] Cheng W,Xie Y.Combined effect of polystyrene microplastics and bisphenol A on the human embryonic stem cells-derived liver organoids: the hepatotoxicity and lipid accumulation.Sci Total Environ2023;854:158585

[54] McCarron S,Conlon DM.Functional characterization of organoids derived from irreversibly damaged liver of patients with NASH.Hepatology2021;74:1825-44

[55] Yildiz E,Perino A.Hepatic lipid overload triggers biliary epithelial cell activation via E2Fs.Elife2023;12:e81926 PMCID:PMC10030116

[56] Wang L,Yu B.Recapitulating lipid accumulation and related metabolic dysregulation in human liver-derived organoids.J Mol Med2022;100:471-84

[57] Chen J,Liu Z.Hepatic glycogenesis antagonizes lipogenesis by blocking S1P via UDPG.Science2024;383:eadi3332

[58] Kømurcu KS,Thorne JL.Mass spectrometry reveals that oxysterols are secreted from non-alcoholic fatty liver disease induced organoids.J Steroid Biochem Mol Biol2023;232:106355

[59] Elbadawy M,Yamamoto H.Evaluation of the efficacy of mitochondrial fission inhibitor (Mdivi-1) using non-alcoholic steatohepatitis (NASH) liver organoids.Front Pharmacol2023;14:1243258 PMCID:PMC10600465

[60] Huang HK,Xu CF.Pre-MASLD: should it be defined separately?.Hepatobiliary Pancreat Dis Int2024;23:1-3