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Cryptogenic cholestasis in young and adults: ATP8B1, ABCB11, ABCB4, and TJP2 gene variants analysis by high-throughput sequencing

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Mutations in ATP-transporters ATPB81, ABCB11, and ABCB4 are responsible for progressive familial intrahepatic cholestasis (PFIC) 1, 2 and 3, and recently the gene for tight junction protein-2 (TJP2) has been linked to PFIC4.

Aim

As these four genes have been poorly studied in young people and adults, we investigated them in this context here.

Methods

In patients with cryptogenic cholestasis, we analyzed the presence of mutations by high-throughput sequencing. Bioinformatics analyses were performed for mechanistic and functional predictions of their consequences on biomolecular interaction interfaces.

Results

Of 108 patients, 48 whose cause of cholestasis was not established were submitted to molecular analysis. Pathogenic/likely pathogenic mutations were found in ten (21%) probands for 13 mutations: two in ATP8B 1, six in ABCB11, two in ABCB4, three in TJP2. We also identified seven variants of uncertain significance: two in ATP8B1, one in ABCB11, two in ABCB4 and two in TJP2. Finally, we identified 11 benign/likely benign variants. Patients with pathogenic/likely pathogenic mutations had higher levels of liver stiffness (measured by FibroScan®) and bile acids, as well as higher rates of cholestatic histological features, compared to the patients without at least likely pathogenic mutations. The multivariate analysis showed that itching was the only independent factor associated with disease-causing mutations (OR 5.801, 95% CI 1.244–27.060, p = 0.025).

Conclusions

Mutations in the genes responsible for PFIC may be involved in both young and adults with cryptogenic cholestasis in a considerable number of cases, including in heterozygous status. Diagnosis should always be suspected, particularly in the presence of itching.

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Abbreviations

PFIC:

Progressive familial intrahepatic cholestasis

TJP2:

Tight junction protein-2

FIC1:

Familial intrahepatic cholestasis 1

BSEP:

Bile salt export pump

MDR:

Multidrug resistance P-glycoprotein 3

GGT:

Gamma-glutamyl-transpeptidase

AP:

Alkaline phosphatase

BRIC:

Benign intrahepatic cholestasis

LPAC:

Low-phospholipid-associated cholelithiasis

ICP:

Intrahepatic cholestasis of pregnancy

DIC:

Drug-induced cholestasis

HTS:

High-throughput sequencing

NGS:

Next-generation sequencing

PSC:

Primary sclerosing cholangitis

BA:

Bile acids

MAF:

Minor allele frequency

SIFT:

Sorting Intolerant From Tolerant

HGMD:

Human Gene Mutation Database

ACMG:

American College of Medical Genetics and Genomics

P:

Pathogenic

LP:

Likely pathogenic

VUS:

Variants of uncertain significance

LB:

Likely benign

B:

Benign

SD:

Standard deviation

CI:

Confidence interval

SNP:

Single-nucleotide polymorphism

ALT:

Alanine aminotransferase

OR:

Odds ratio

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Author information

Authors and Affiliations

  1. Department of Medical and Surgical Sciences, University of Bologna, Bologna, Italy

    Giovanni Vitale, Stefano Gitto, Ranka Vukotic, Marco Seri & Pietro Andreone

  2. Center for Applied Biomedical Research (CRBA), University Hospital, Bologna, Italy

    Alessandro Mattiaccio & Vilma Mantovani

  3. CellNetworks, Bioquant, Heidelberg University, Im Neuenheimer Feld 267, 69120, Heidelberg, Germany

    Francesco Raimondi & Robert B. Russell

  4. Bioochemie Zentrum Heidelberg (BZH), Heidelberg University, Im Neuenheimer Feld 328, 69120, Heidelberg, Germany

    Francesco Raimondi & Robert B. Russell

  5. Addari Institute of Oncology and Transplant Pathology, Policlinico S. Orsola-Malpighi, University of Bologna, Bologna, Italy

    Antonietta D’Errico

  6. Department of Medical and Surgical Sciences and Research Center for the Study of Hepatitis, University of Bologna, Italy, Via Massarenti 9, 40138, Bologna, Italy

    Pietro Andreone

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  1. Giovanni Vitale
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Contributions

GV and PA designed the study and collected data. AM, VM, and MS performed the DNA sequencing and applied prediction tools; AD supervised the histological evaluations, FR and RBR performed protein modeling by Mechismo; SG, AM, VM, GV, RV, and PA analyzed the patients’ data. GV wrote the manuscript; all authors critically revised the manuscript.

Corresponding author

Correspondence to Pietro Andreone.

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The authors declare that they have no conflict of interest.

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Vitale, G., Gitto, S., Raimondi, F. et al. Cryptogenic cholestasis in young and adults: ATP8B1, ABCB11, ABCB4, and TJP2 gene variants analysis by high-throughput sequencing. J Gastroenterol 53, 945–958 (2018). https://doi.org/10.1007/s00535-017-1423-1

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  • DOI: https://doi.org/10.1007/s00535-017-1423-1

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