Diagnosis of ABCB11 gene mutations in children with intrahepatic cholestasis using high resolution melting analysis and direct sequencing

Hu,Guorui; He,Ping; Liu,Zhifeng; Chen,Qian; Zheng,Bixia; Zhang,Qihua

doi:10.3892/mmr.2014.2349

Diagnosis of ABCB11 gene mutations in children with intrahepatic cholestasis using high resolution melting analysis and direct sequencing

Authors:
- Guorui Hu
- Ping He
- Zhifeng Liu
- Qian Chen
- Bixia Zheng
- Qihua Zhang
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Affiliations: Medical College of Nanjing University, Nanjing, Jiangsu 210093, P.R. China, Department of Digestive Disease, Nanjing Children's Hospital, Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China
Published online on: June 20, 2014 https://doi.org/10.3892/mmr.2014.2349
Pages: 1264-1274
Copyright: © Hu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Intrahepatic cholestasis represents a heterogeneous group of disorders that begin during childhood, most commonly manifesting as neonatal cholestasis, and lead to ongoing liver dysfunction in children and adults. For children, inherited pathogenic factors of cholestasis have gained increasing attention owing to the rapid development of molecular biology technology. However, these methods have their advantages and disadvantages in terms of simplicity, sensitivity, specificity, time required and expense. In the present study, an effective, sensitive and economical method is recommended, termed high-resolution melting (HRM) analysis and direct sequencing, based on general polymerase chain reaction, to detect mutations in disease‑causing genes. As one type of inherited intrahepatic cholestasis, progressive familial intrahepatic cholestasis type 2 (PFIC2) is caused by pathogenic mutations in the ABCB11 gene, HRM was used to detect mutations in the ABCB11 gene in the present study, and the diagnosis for PFIC2 was made by comprehensive analysis of genetic findings and clinical features. Furthermore, the characteristics of mutations and single nucleotide polymorphisms (SNPs) in the ABCB11 gene were elucidated. A total of 14 types of mutations/polymorphisms were identified in 20 patients from mainland China, including six missense mutations (p.Y337H, p.Y472C, p.R696W, p.Q931P, p.D1131V and p.H1198R), one nonsense mutation (p.R928X) and seven SNPs (p.D36D/rs3815675, p.F90F/rs4148777, p.Y269Y/rs2287616, p.I416I/rs183390670, p.V444A/rs2287622, p.A865V/rs118109635 and p.A1028A/rs497692). Five mutations were novel. The majority of the mutations were different from those detected in other population groups. A total of 4/20 patients (1/5) were diagnosed to be PFIC2 by combining genetic findings with the clinical features. Polymorphisms V444A and A1028A, with an allele frequency of 74.5 and 67.2%, respectively, were highly prevalent in the mainland Chinese subjects. No differences were found between the patients with cholestasis and the control subjects. Efficient genetic screening facilitates the clinical diagnosis of genetic disorders. The present study demonstrated that HRM analysis was efficient and effective in detecting mutations and expanded the known spectrum of ABCB11 gene mutations.

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1	Keitel V, Burdelski M, Vojnisek Z, Schmitt L, Häussinger D and Kubitz R: De novo bile salt transporter antibodies as a possible cause of recurrent graft failure after liver transplantation: a novel mechanism of cholestasis. Hepatology. 50:510–517. 2009. View Article : Google Scholar : PubMed/NCBI
2	Morotti RA, Suchy FJ and Magid MS: Progressive familial intrahepatic cholestasis (PFIC) type 1, 2, and 3: a review of the liver pathology findings. Semin Liver Dis. 31:3–10. 2011. View Article : Google Scholar : PubMed/NCBI
3	Harris MJ, Le Couteur DG and Arias IM: Progressive familial intrahepatic cholestasis: genetic disorders of biliary transporters. J Gastroenterol Hepatol. 20:807–817. 2005. View Article : Google Scholar : PubMed/NCBI
4	Hori T, Nguyen JH and Uemoto S: Progressive familial intrahepatic cholestasis. Hepatobiliary Pancreat Dis Int. 9:570–578. 2010.
5	Nicolaou M, Andress EJ, Zolnerciks JK, Dixon PH, Williamson C and Linton KJ: Canalicular ABC transporters and liver disease. J Pathol. 226:300–315. 2012. View Article : Google Scholar : PubMed/NCBI
6	Jankowska I and Socha P: Progressive familial intrahepatic cholestasis and inborn errors of bile acid synthesis. Clin Res Hepatol Gastroenterol. 36:271–274. 2012. View Article : Google Scholar : PubMed/NCBI
7	Strautnieks SS, Bull LN, Knisely AS, et al: A gene encoding a liver-specific ABC transporter is mutated in progressive familial intrahepatic cholestasis. Nat Genet. 20:233–238. 1998. View Article : Google Scholar : PubMed/NCBI
8	van Mil SW, van der Woerd WL, van der Brugge G, et al: Benign recurrent intrahepatic cholestasis type 2 is caused by mutations in ABCB11. Gastroenterology. 127:379–384. 2004.PubMed/NCBI
9	Andrade RJ, Robles M, Ulzurrun E and Lucena MI: Drug-induced liver injury: insights from genetic studies. Pharmacogenomics. 10:1467–1487. 2009. View Article : Google Scholar : PubMed/NCBI
10	Dixon PH, van Mil SW, Chambers J, et al: Contribution of variant alleles of ABCB11 to susceptibility to intrahepatic cholestasis of pregnancy. Gut. 58:537–544. 2009. View Article : Google Scholar : PubMed/NCBI
11	Meier Y, Zodan T, Lang C, et al: Increased susceptibility for intrahepatic cholestasis of pregnancy and contraceptive-induced cholestasis in carriers of the 1331T>C polymorphism in the bile salt export pump. World J Gastroenterol. 14:38–45. 2008. View Article : Google Scholar : PubMed/NCBI
12	Stieger B and Geier A: Genetic variations of bile salt transporters as predisposing factors for drug-induced cholestasis, intrahepatic cholestasis of pregnancy and therapeutic response of viral hepatitis. Expert Opin Drug Metab Toxicol. 7:411–425. 2011. View Article : Google Scholar
13	Müllenbach R, Weber SN, Krawczyk M, et al: A frequent variant in the human bile salt export pump gene ABCB11 is associated with hepatitis C virus infection, but not liver stiffness in a German population. BMC Gastroenterol. 12:632012.
14	Byrne JA, Strautnieks SS, Ihrke G, et al: Missense mutations and single nucleotide polymorphisms in ABCB11 impair bile salt export pump processing and function or disrupt pre-messenger RNA splicing. Hepatology. 49:553–567. 2009. View Article : Google Scholar
15	Pauli-Magnus C, Kerb R, Fattinger K, et al: BSEP and MDR3 haplotype structure in healthy Caucasians, primary biliary cirrhosis and primary sclerosing cholangitis. Hepatology. 39:779–791. 2004. View Article : Google Scholar : PubMed/NCBI
16	Chen HL, Liu YJ, Su YN, et al: Diagnosis of BSEP/ABCB11 mutations in Asian patients with cholestasis using denaturing high performance liquid chromatography. J Pediatr. 153:825–832. 2008. View Article : Google Scholar : PubMed/NCBI
17	Liu LY, Wang ZL, Wang XH, Zhu QR and Wang JS: ABCB11 gene mutations in Chinese children with progressive intrahepatic cholestasis and low gamma glutamyltransferase. Liver Int. 30:809–815. 2010. View Article : Google Scholar
18	Nguyen-Dumont T, Calvez-Kelm FL, Forey N, et al: Description and validation of high-throughput simultaneous genotyping and mutation scanning by high-resolution melting curve analysis. Hum Mutat. 30:884–890. 2009. View Article : Google Scholar : PubMed/NCBI
19	Chen HL, Liu YJ, Su YN, et al: Diagnosis of BSEP/ABCB11 mutations in Asian patients with cholestasis using denaturing high performance liquid chromatograph. J Pediatr. 153:825–832. 2008. View Article : Google Scholar : PubMed/NCBI
20	Schwarz JM, Rödelsperger C, Schuelke M and Seelow D: MutationTaster evaluates disease-causing potential of sequence alterations. Nat Methods. 7:575–576. 2010. View Article : Google Scholar : PubMed/NCBI
21	Ng PC and Henikoff S: SIFT: Predicting amino acid changes that affect protein function. Nucleic Acids Res. 31:3812–3814. 2003. View Article : Google Scholar : PubMed/NCBI
22	Castellana S and Mazza T: Congruency in the prediction of pathogenic missense mutations: state-of-the-art web-based tools. Brief Bioinform. 14:448–459. 2013. View Article : Google Scholar : PubMed/NCBI
23	Adzhubei I, Jordan DM and Sunyaev SR: 2013.Predicting Functional Effect of Human Missense Mutations Using PolyPhen-2. Curr Protocols in Human Genetics. 76:7.20.1–7.20.41
24	Calabrese R, Capriotti E, Fariselli P, Martelli PL and Casadio R: Functional annotations improve the predictive score of human disease-related mutations in proteins. Hum Mutat. 30:1237–1244. 2009. View Article : Google Scholar : PubMed/NCBI
25	Leabman MK, Huang CC, DeYoung J, et al: Natural variation in human membrane transporter genes reveals evolutionary and functional constraints. Proc Natl Acad Sci USA. 100:5896–5901. 2003. View Article : Google Scholar : PubMed/NCBI
26	Evason K, Bove KE, Finegold MJ, et al: Morphologic findings in progressive familial intrahepatic cholestasis 2 (PFIC2): correlation with genetic and immunohistochemical studies. Am J Surg Pathol. 35:687–696. 2011. View Article : Google Scholar
27	Strautnieks SS, Byrne JA, Pawlikowska L, et al: Severe bile salt export pump deficiency: 82 different ABCB11 mutations in 109 families. Gastroenterology. 134:1203–1214. 2008. View Article : Google Scholar : PubMed/NCBI
28	Liu C, Aronow BJ, Jegga AG, et al: Novel resequencing chip customized to diagnose mutations in patients with inherited syndromes of intrahepatic cholestasis. Gastroenterology. 132:119–126. 2007. View Article : Google Scholar : PubMed/NCBI
29	Acalovschi M, Tirziu S, Chiorean E, Krawczyk M, Grünhage F and Lammert F: Common variants of ABCB4 and ABCB11 and plasma lipid levels: a study in sib pairs with gallstones, and controls. Lipids. 44:521–526. 2009. View Article : Google Scholar : PubMed/NCBI
30	Wheeler DA, Srinivasan M, Egholm M, et al: The complete genome of an individual by massively parallel DNA sequencing. Nature. 452:872–876. 2008. View Article : Google Scholar : PubMed/NCBI
31	van der Stoep N, van Paridon CD, Janssens T, et al: Diagnostic guidelines for high-resolution melting curve (HRM) analysis: an interlaboratory validation of BRCA1 mutation scanning using the 96-well LightScanner. Hum Mutat. 30:899–909. 2009.PubMed/NCBI
32	van der Stoep N, van Paridon CD, Janssens T, et al: Diagnostic guidelines for high-resolution melting curve (HRM) analysis: an interlaboratory validation of BRCA1 mutation scanning using the 96-well LightScanner. Hum Mutat. 30:899–909. 2009.PubMed/NCBI
33	Zolnerciks JK, Andress EJ, Nicolaou M and Linton KJ: Structure of ABC transporters. Essays Biochem. 50:43–61. 2011. View Article : Google Scholar : PubMed/NCBI
34	Matte U, Mourya R, Miethke A, et al: Analysis of gene mutations in children with cholestasis of undefined etiology. J Pediatr Gastroenterol Nutr. 51:488–493. 2010. View Article : Google Scholar : PubMed/NCBI
35	Treepongkaruna S, Gaensan A, Pienvichit P, et al: Novel ABCB11 mutations in a Thai infant with progressive familial intrahepatic cholestasis. World J Gastroenterol. 15:4339–4342. 2009. View Article : Google Scholar : PubMed/NCBI
36	Nobili V, Di Giandomenico S, Francalanci P, Callea F, Marcellini M and Santorelli FM: A new ABCB11 mutation in two Italian children with familial intrahepatic cholestasis. J Gastroenterol. 41:598–603. 2006. View Article : Google Scholar : PubMed/NCBI
37	Goto K, Sugiyama K, Sugiura T, et al: Bile salt export pump gene mutations in two Japanese patients with progressive familial intrahepatic cholestasis. J Pediatr Gastroenterol Nutr. 36:647–650. 2003. View Article : Google Scholar : PubMed/NCBI
38	Saito S, Iida A, Sekine A, et al: Three hundred twenty-six genetic variations in genes encoding nine members of ATP-binding cassette, subfamily B (ABCB/MDR/TAP), in the Japanese population. J Hum Genet. 47:38–50. 2002. View Article : Google Scholar
39	Kim SR, Saito Y, Itoda M, et al: Genetic variations of the ABC transporter gene ABCB11 encoding the human bile salt export pump (BSEP) in a Japanese population. Drug Metab Pharmacokinet. 24:277–281. 2009. View Article : Google Scholar : PubMed/NCBI
40	Chen HL, Chang PS, Hsu HC, et al: FIC1 and BSEP defects in Taiwanese patients with chronic intrahepatic cholestasis with low gamma-glutamyltranspeptidase levels. J Pediatr. 140:119–124. 2002. View Article : Google Scholar : PubMed/NCBI
41	Chen ST, Chen HL, Su YN, et al: Prenatal diagnosis of progressive familial intrahepatic cholestasis type 2. J Gastroenterol Hepatol. 23:1390–1393. 2008. View Article : Google Scholar : PubMed/NCBI
42	Liu LY, Wang XH, Lu Y, Zhu QR and Wang JS: Association of variants of ABCB11 with transient neonatal cholestasis. Pediatr Int. 55:138–144. 2013. View Article : Google Scholar : PubMed/NCBI
43	Ananthanarayanan M and Li Y: PFIC2 and ethnicity-specific bile salt export pump (BSEP, ABCB11) mutations: where do we go from here? Liver Int. 30:777–779. 2010. View Article : Google Scholar : PubMed/NCBI
44	Lang T, Haberl M, Jung D, et al: Genetic variability, haplotype structures, and ethnic diversity of hepatic transporters MDR3 (ABCB4) and bile salt export pump (ABCB11). Drug Metab Dispos. 34:1582–1599. 2006. View Article : Google Scholar : PubMed/NCBI
45	Lang C, Meier Y, Stieger B, et al: Mutations and polymorphisms in the bile salt export pump and the multidrug resistance protein 3 associated with drug-induced liver injury. Pharmacogenet Genomics. 17:47–60. 2007. View Article : Google Scholar : PubMed/NCBI