SLCO3A1, A novel crohn's disease-associated gene, regulates nf-κB activity and associates with intestinal perforation

Shu-Chen Wei; Yan-Yin Tan; Meng-Tzu Weng; Liang-Chuan Lai; Jen-Hao Hsiao; Eric Y Chuang; Chia-Tung Shun; Deng-Cheng Wu; Ai-Wen Kao; Chiao-Shung Chuang; Yen-Hsuan Ni; Ming-Jium Shieh; Chien-Chih Tung; Yun Chen; Cheng-Yi Wang; Ramnik J Xavier; Daniel K Podolsky; Jau-Min Wong

doi:10.1371/journal.pone.0100515

SLCO3A1, A novel crohn's disease-associated gene, regulates nf-κB activity and associates with intestinal perforation

PLoS One. 2014 Jun 19;9(6):e100515. doi: 10.1371/journal.pone.0100515. eCollection 2014.

Authors

Shu-Chen Wei¹, Yan-Yin Tan², Meng-Tzu Weng³, Liang-Chuan Lai⁴, Jen-Hao Hsiao⁵, Eric Y Chuang⁶, Chia-Tung Shun⁷, Deng-Cheng Wu⁸, Ai-Wen Kao⁹, Chiao-Shung Chuang⁹, Yen-Hsuan Ni¹⁰, Ming-Jium Shieh¹¹, Chien-Chih Tung¹², Yun Chen¹³, Cheng-Yi Wang¹, Ramnik J Xavier¹⁴, Daniel K Podolsky¹⁵, Jau-Min Wong¹⁶

Affiliations

¹ Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan.
² Graduate Institute of Medical Engineering, National Taiwan University, Taipei, Taiwan.
³ Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan; Departments of Internal Medicine, Far Eastern Memorial Hospital, New Taipei, Taiwan.
⁴ Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei, Taiwan; Bioinformatics and Biostatistics Core, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan.
⁵ Bioinformatics and Biostatistics Core, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan.
⁶ Bioinformatics and Biostatistics Core, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan; Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, Taiwan.
⁷ Department of Pathology and Forensic Medicine, National Taiwan University Hospital, Taipei, Taiwan.
⁸ Department of Internal Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
⁹ Department of Internal Medicine, National Cheng Kung University, Tainan, Taiwan.
¹⁰ Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan.
¹¹ Graduate Institute of Medical Engineering, National Taiwan University, Taipei, Taiwan; Deparment of Oncology, National Taiwan University Hospital, Taipei, Taiwan.
¹² Department of Integrated Diagnostics and Therapeutics, National Taiwan University Hospital, Taipei, Taiwan.
¹³ Pediatric Surgery, Far Eastern Memorial Hospital, New Taipei, Taiwan.
¹⁴ Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
¹⁵ UT Southwestern Medical Center, Dallas, Texas, United States of America.
¹⁶ Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan; Graduate Institute of Medical Engineering, National Taiwan University, Taipei, Taiwan.

Abstract

Background & aims: To date, only one gene (TNFSF15) has been identified and validated as a Crohn's disease (CD)-associated gene in non-Caucasian populations. This study was designed to identify novel CD-associated single nucleotide polymorphisms (SNPs)/genes and to validate candidate genes using a functional assay.

Methods: SNPs from 16 CD patients and 16 age- and sex-matched control patients were analyzed using Illumina platform analysis. Subsequently, we expanded the study and followed 53 CD patients and 41 control patients by Sequenom MassArray analysis. Quantitative PCR and immunohistochemical staining were performed to assess mRNA and protein expression of the candidate gene on tissue isolated from CD patients. Genotype was correlated with CD phenotypes. Finally, the candidate gene was cloned and its effect on NF-κB activity assessed using a reporter luciferase assay.

Results: SLCO3A1 (rs207959) reached statistical significance in the first-stage analysis (P = 2.3E-02) and was further validated in the second-stage analysis (P = 1.0E-03). Genotype and phenotype analysis showed that the rs207959 (T) allele is a risk allele that alters SLCO3A1 mRNA expression and is associated with intestinal perforation in CD patients. Higher levels of mRNA and protein expression of SLCO3A1 were seen in CD patients compared with the control group. Overexpression of SLCO3A1 induced increased NF-κB activity and increased phosphorylation of P65, ERK, and JNK. Nicotine augmented the activation of NF-κB in the presence of SLCO3A1.

Conclusions: SLCO3A1, a novel CD-associated gene, mediates inflammatory processes in intestinal epithelial cells through NF-κB transcription activation, resulting in a higher incidence of bowel perforation in CD patients.

MeSH terms

Adolescent
Adult
Aged
Alleles
Case-Control Studies
Child
Crohn Disease / complications
Crohn Disease / enzymology
Crohn Disease / genetics*
Demography
Female
Genetic Association Studies*
Genetic Predisposition to Disease*
HEK293 Cells
Humans
Intestinal Perforation / complications
Intestinal Perforation / genetics*
Intestinal Perforation / pathology
Male
Middle Aged
Mitogen-Activated Protein Kinases / metabolism
NF-kappa B / metabolism*
Nicotine / pharmacology
Organic Anion Transporters / genetics*
Organic Anion Transporters / metabolism
Phosphorylation / drug effects
Polymorphism, Single Nucleotide / genetics
Proto-Oncogene Proteins c-akt / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism
Young Adult

Substances

NF-kappa B
Organic Anion Transporters
RNA, Messenger
SLCO3A1 protein, human
Nicotine
Proto-Oncogene Proteins c-akt
Mitogen-Activated Protein Kinases

Grants and funding

This work was supported by National Science Council of Taiwan (NSC-98-2314-B-002-139-) (99-2314-B-002-124-MY3); Liver Disease Prevention & Treatment Research Foundation, Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.