Ningappa, M and So, J and Glessner, J and Ashokkumar, C and Ranganathan, S and Min, J and Higgs, BW and Sun, Q and Haberman, K and Schmitt, L and Vilarinho, S and Mistry, PK and Vockley, G and Dhawan, A and Gittes, GK and Hakonarson, H and Jaffe, R and Subramaniam, S and Shin, D and Sindhi, R
(2015)
The role of ARF6 in biliary atresia.
PLoS ONE, 10 (9).
Abstract
Background & Aims: Altered extrahepatic bile ducts, gut, and cardiovascular anomalies constitute the variable phenotype of biliary atresia (BA). Methods: To identify potential susceptibility loci, Caucasian children, normal (controls) and with BA (cases) at two US centers were compared at >550000 SNP loci. Systems biology analysis was carried out on the data. In order to validate a key gene identified in the analysis, biliary morphogenesis was evaluated in 2-5-day post-fertilization zebrafish embryos after morpholino-antisense oligonucleotide knockdown of the candidate gene ADP ribosylation factor-6 (ARF6, Mo-arf6). Results: Among 39 and 24 cases at centers 1 and 2, respectively, and 1907 controls, which clustered together on principal component analysis, the SNPs rs3126184 and rs10140366 in a 3' flanking enhancer region for ARF6 demonstrated higher minor allele frequencies (MAF) in each cohort, and 63 combined cases, compared with controls (0.286 vs. 0.131, P = 5.94 x 10-7, OR 2.66; 0.286 vs. 0.13, P = 5.57 x 10-7, OR 2.66). Significance was enhanced in 77 total cases, which included 14 additional BA genotyped at rs3126184 only (p = 1.58 x 10-2, OR = 2.66). Pathway analysis of the 1000 top-ranked SNPs in CHP cases revealed enrichment of genes for EGF regulators (p<1 x 10-7), ERK/MAPK and CREB canonical pathways (p<1 x 10-34), and functional networks for cellular development and proliferation (p<1 x 10-45), further supporting the role of EGFR-ARF6 signaling in BA. In zebrafish embryos, Mo-arf6 injection resulted in a sparse intrahepatic biliary network, several biliary epithelial cell defects, and poor bile excretion to the gall bladder compared with uninjected embryos. Biliary defects were reproduced with the EGFR-blocker AG1478 alone or with Mo-arf6 at lower doses of each agent and rescued with arf6 mRNA. Conclusions: The BA-associated SNPs identify a chromosome 14q21.3 susceptibility locus encompassing the ARF6 gene. arf6 knockdown in zebrafish implicates early biliary dysgenesis as a basis for BA, and also suggests a role for EGFR signaling in BA pathogenesis.
Share
Citation/Export: |
|
Social Networking: |
|
Details
Item Type: |
Article
|
Status: |
Published |
Creators/Authors: |
Creators | Email | Pitt Username | ORCID |
---|
Ningappa, M | | | | So, J | juhoon@pitt.edu | JUHOON | | Glessner, J | | | | Ashokkumar, C | csa3@pitt.edu | CSA3 | | Ranganathan, S | sar1@pitt.edu | SAR1 | 0000-0001-6214-9167 | Min, J | | | | Higgs, BW | | | | Sun, Q | qingsun@pitt.edu | QINGSUN | | Haberman, K | | | | Schmitt, L | | | | Vilarinho, S | | | | Mistry, PK | | | | Vockley, G | jerryvockley@pitt.edu | GEV1 | 0000-0002-8180-6457 | Dhawan, A | | | | Gittes, GK | gkg2@pitt.edu | GKG2 | | Hakonarson, H | | | | Jaffe, R | | | | Subramaniam, S | | | | Shin, D | donghuns@pitt.edu | DONGHUNS | | Sindhi, R | | | |
|
Contributors: |
Contribution | Contributors Name | Email | Pitt Username | ORCID |
---|
Editor | Alpini, Gianfranco | UNSPECIFIED | UNSPECIFIED | UNSPECIFIED |
|
Centers: |
Other Centers, Institutes, Offices, or Units > McGowan Institute for Regenerative Medicine |
Date: |
17 September 2015 |
Date Type: |
Publication |
Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
Journal or Publication Title: |
PLoS ONE |
Volume: |
10 |
Number: |
9 |
DOI or Unique Handle: |
10.1371/journal.pone.0138381 |
Institution: |
University of Pittsburgh |
Schools and Programs: |
School of Medicine > Pathology School of Medicine > Pediatrics |
Refereed: |
Yes |
Date Deposited: |
23 Aug 2016 14:38 |
Last Modified: |
04 Feb 2023 11:55 |
URI: |
http://d-scholarship.pitt.edu/id/eprint/28385 |
Metrics
Monthly Views for the past 3 years
Plum Analytics
Altmetric.com
Actions (login required)
|
View Item |