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Journal of Gastroenterology

, Volume 53, Issue 8, pp 945–958 | Cite as

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

  • Giovanni Vitale
  • Stefano Gitto
  • Francesco Raimondi
  • Alessandro Mattiaccio
  • Vilma Mantovani
  • Ranka Vukotic
  • Antonietta D’Errico
  • Marco Seri
  • Robert B. Russell
  • Pietro Andreone
Original Article—Liver, Pancreas, and Biliary Tract

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.

Keywords

Progressive familial intrahepatic cholestasis Cryptogenic disease Pathogenic mutations Genetic variants Bioinformatics analysis 

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

Notes

Author 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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Financial support

No grants and other financial support were received.

Supplementary material

535_2017_1423_MOESM1_ESM.pdf (6 kb)
Supplementary material 1 (PDF 5 kb)
535_2017_1423_MOESM2_ESM.docx (25 kb)
Supplementary material 2 (DOCX 25 kb)
535_2017_1423_MOESM3_ESM.xlsx (20 kb)
Supplementary material 3 (XLSX 20 kb)
535_2017_1423_MOESM4_ESM.doc (52 kb)
Supplementary material 4 (DOC 51 kb)

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Copyright information

© Japanese Society of Gastroenterology 2017

Authors and Affiliations

  • Giovanni Vitale
    • 1
  • Stefano Gitto
    • 1
  • Francesco Raimondi
    • 3
    • 4
  • Alessandro Mattiaccio
    • 2
  • Vilma Mantovani
    • 2
  • Ranka Vukotic
    • 1
  • Antonietta D’Errico
    • 5
  • Marco Seri
    • 1
  • Robert B. Russell
    • 3
    • 4
  • Pietro Andreone
    • 1
    • 6
  1. 1.Department of Medical and Surgical SciencesUniversity of BolognaBolognaItaly
  2. 2.Center for Applied Biomedical Research (CRBA)University HospitalBolognaItaly
  3. 3.CellNetworks, BioquantHeidelberg UniversityHeidelbergGermany
  4. 4.Bioochemie Zentrum Heidelberg (BZH)Heidelberg UniversityHeidelbergGermany
  5. 5.Addari Institute of Oncology and Transplant Pathology, Policlinico S. Orsola-MalpighiUniversity of BolognaBolognaItaly
  6. 6.Department of Medical and Surgical Sciences and Research Center for the Study of HepatitisUniversity of Bologna, ItalyBolognaItaly

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