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Hepatol Int. 2019 Dec 13. doi: 10.1007/s12072-019-10007-y. [Epub ahead of print]

Liver organoids reproduce alpha-1 antitrypsin deficiency-related liver disease.

Author information

1
Molecular Genetics Unit, Institute of Rare Diseases Research, Institute of Health Carlos III (ISCIII), Ctra. Majadahonda-Pozuelo Km2,200, 28220, Madrid, Spain.
2
General and Digestive Surgery Department, Hospital Doce de Octubre, Madrid, Spain.
3
Bioinformatics Unit, Institute of Health Carlos III (ISCIII), Madrid, Spain.
4
Digestive Department, Hospital Doce de Octubre, Madrid, Spain.
5
Neumology Department, Hospital Doce de Octubre, Madrid, Spain.
6
Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK.
7
Gene Therapy Unit, Institute of Rare Diseases Research, Institute of Health Carlos III (ISCIII), Madrid, Spain.
8
Institute of Rare Diseases Research, Institute of Health Carlos III (ISCIII), Centre for Biomedical Network Research on Rare Diseases, CIBERER, Madrid, Spain.
9
Department of Respiratory Medicine, German Centre for Lung Research (DZL), Hannover Medical School, Hannover, Germany.
10
Molecular Genetics Unit, Institute of Rare Diseases Research, Institute of Health Carlos III (ISCIII), Ctra. Majadahonda-Pozuelo Km2,200, 28220, Madrid, Spain. bmartinezd@isciii.es.
11
Institute of Rare Diseases Research, Institute of Health Carlos III (ISCIII), Centre for Biomedical Network Research on Rare Diseases, CIBERER, Madrid, Spain. bmartinezd@isciii.es.

Abstract

BACKGROUND AND AIMS:

Alpha-1 antitrypsin (AAT) is a product of SERPINA1 gene mainly expressed by hepatocytes. Clinically relevant mutations in the SERPINA1 gene, such as Z (Glu342Lys), results in an expression of misfolded AAT protein having high propensity to polymerize, accumulate in hepatocytes and thus to enhance a risk for hepatocyte damage and subsequent liver disease. So far, the relationship between the Z-AAT accumulation and liver cell damage remains not completely understood. We present three-dimensional organoid culture systems, as a novel tool for modeling Z-AAT-related liver diseases.

METHODS:

We have established liver organoids from liver biopsies of patients with homozygous (ZZ) and heterozygous (MZ) deficiency and normal (MM) genotypes of AAT. The features of these organoid models were characterized by analyzing AAT protein secretion and intracellular aggregation in MZ and ZZ genotypes as well as SERPINA1 expression in differentiated cultures.

RESULTS:

Transcriptional analysis of differentiated organoid cultures by RNA-Seq showed hepatocyte-specific gene expression profile. Genes, such as ALB, APOB, CYP3A4 and SERPINA1, were validated and confirmed through quantitative-PCR analysis. The organoids from MZ and ZZ cases showed intracellular aggregation and lower secretion of AAT protein, and lower expression of ALB and APOB, as typically seen in hepatocytes from Z-AAT deficiency patients. Furthermore, organoids responded to external stimulus. Treatment with oncostatin M, a well-known inducer of SERPINA1, increased expression of the full-length transcripts (AAT-1C) as well as the short transcript of AAT (AAT-ST1C4).

CONCLUSIONS:

Liver organoid model recapitulates the key features of Z-AAT deficiency and provides a useful tool for disease modeling.

KEYWORDS:

Alpha-1 antitrypsin deficiency; Liver; Oncostatin M; Organoids; RNA-seq; SERPINA1

PMID:
31832977
DOI:
10.1007/s12072-019-10007-y

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