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Cited by 98Pediatric Hepatobiliary Diseases and TreatmentsLiver physiology and pathologyDrug Transport and Resistance Mechanisms

Biliary organoids uncover delayed epithelial development and barrier function in biliary atresia

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Amarachintha, Surya P., Mourya, Reena, Ayabe, Hiroaki, Yang, Li, Luo, Zhenhua, Li, Xiaofeng, Thanekar, Unmesha, Shivakumar, Pranavkumar +1 more·2022·Hepatology
DOI 10.1002/hep.32107PMC PMC9983428
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Abstract

Background and Aims: Biliary atresia is a severe inflammatory and fibrosing cholangiopathy of neonates of unknown etiology. The onset of cholestasis at birth implies a prenatal onset of liver dysfunction. Our aim was to investigate the mechanisms linked to abnormal cholangiocyte development. Approach and Results: We generated biliary organoids from liver biopsies of infants with biliary atresia and normal and diseased controls. Organoids emerged from biliary atresia livers and controls and grew as lumen‐containing spheres with an epithelial lining of cytokeratin‐19 pos albumin neg SOX17 neg cholangiocyte‐like cells. Spheres had similar gross morphology in all three groups and expressed cholangiocyte‐enriched genes. In biliary atresia, cholangiocyte‐like cells lacked a basal positioning of the nucleus, expressed fewer developmental and functional markers, and displayed misorientation of cilia. They aberrantly expressed F‐actin, β‐catenin, and Ezrin, had low signals for the tight junction protein zonula occludens‐1 (ZO‐1), and displayed increased permeability as evidenced by a higher Rhodamine‐123 (R123) signal inside organoids after verapamil treatment. Biliary atresia organoids had decreased expression of genes related to EGF signaling and FGF2 signaling. When treated with EGF+FGF2, biliary atresia organoids expressed differentiation (cytokeratin 7 and hepatocyte nuclear factor 1 homeobox B) and functional (somatostatin receptor 2, cystic fibrosis transmembrane conductance regulator [CFTR], aquaporin 1) markers, restored polarity with improved localization of F‐actin, β‐catenin and ZO‐1, increased CFTR function, and decreased uptake of R123. Conclusions: Organoids from biliary atresia are viable and have evidence of halted epithelial development. The induction of developmental markers, improved cell‐cell junction, and decreased epithelial permeability by EGF and FGF2 identifies potential strategies to promote epithelial maturation and function.

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  • Liver organoids derived from LGR5-positive liver progenitor cells can be stably maintained for months and can differentiate into functionally competent hepatocytes and cholangiocytes.

    “Liver organoids derived from leucine-rich repeat–containing G protein–coupled receptor 5 (LGR5)–positive liver progenitor cells (LPC) can be stably maintained for months and can differentiate into functionally competent hepatocytes and cholangiocytes [14–16].”

    From High-throughput human microfluidic organoid-on-a-chip platform for modeling liver diseases and screening nanotherapeutics. (2026) · cites this paper as [16]

    The Results section on EGF/FGF-induced differentiation reports improved cholangiocyte markers and a gain in chloride-transport function in differentiated biliary-atresia organoids. The paper does not establish that these organoids came from LGR5-positive progenitors, were stably maintained for months, or differentiated into functional hepatocytes; its Methods describe derivation from liver tissue and 14 days of cholangiocyte differentiation. (LLM, conf=0.86)

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Authors
Amarachintha, S. P., Mourya, R., Ayabe, H., Yang, L., Luo, Z., Li, X., Thanekar, U., Shivakumar, P., & Bezerra, J. A.
Affiliations
  • Cincinnati Children's Hospital Medical Center (US)
  • The First Affiliated Hospital, Sun Yat-sen University (CN)
  • Third Affiliated Hospital of Sun Yat-sen University (CN)
Corresponding author
  • Shivakumar, Pranavkumar
  • Bezerra, Jorge A.
ORCID
Venue
Hepatology
Publisher
Ovid Technologies (Wolters Kluwer Health)
Published
2021-12-04
Volume
75
Issue
1
Pages
89-103
ISSN
  • 0270-9139 (ISSN-L)
  • 1527-3350
DOI
10.1002/hep.32107
PMC
PMC9983428
OpenAlex
W3196032083
Cited by
98
References
33
Citations by year
2026: 20 · 2025: 20 · 2024: 25 · 2023: 19 · 2022: 12 · 2021: 2
FWCI
13.62
Type
journal article
Language
en
Open access
Licence
Locations
  • Hepatology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health (7r01dk083781-14)
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK 83781, R01-DK083781, DK78392, DK 64008, R01DK064008, P30DK078392)
Topics
  • Health Sciences › Medicine › Surgery › Pediatric Hepatobiliary Diseases and Treatments
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
biliary atresia, epithelial barrier function, epithelial permeability, tight junctions, zonula occludens-1, EGF signaling, FGF2 signaling, CFTR, epithelial polarity, cholestasis, neonatal liver disease
MeSH terms
Adolescent, Bile Ducts, Biliary Atresia, Biopsy, Cells, Cultured, Child, Child, Preschool, Cholestasis, Epithelial Cells, Humans, Infant, Infant, Newborn, Organoids, Case-Control Studies, Tight Junctions, Primary Cell Culture, Healthy Volunteers

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