The Ser82 RAGE Variant Affects Lung Function and Serum RAGE in Smokers and sRAGE Production In Vitro

Miller, Suzanne; Henry, Amanda P.; Hodge, Emily; Kheirallah, Alexander K.; Billington, Charlotte K.; Rimington, Tracy L.; Bhaker, Sangita K.; Obeidat, Ma�en; Mel�n, Erik; Merid, Simon K.; Swan, Caroline; Gowland, Catherine; Nelson, Carl P.; Stewart, Ceri E.; Bolton, Charlotte E.; Kilty, Iain; Malarstig, Anders; Parker, Stuart G.; Moffatt, Miriam F.; Wardlaw, Andrew J.; Hall, Ian P.; Sayers, Ian

Abstract

Introduction

Genome-Wide Association Studies have identified associations between lung function measures and Chronic Obstructive Pulmonary Disease (COPD) and chromosome region 6p21 containing the gene for the Advanced Glycation End Product Receptor (AGER, encoding RAGE). We aimed to (i) characterise RAGE expression in the lung, (ii) identify AGER transcripts, (iii) ascertain if SNP rs2070600 (Gly82Ser C/T) is associated with lung function and serum sRAGE levels and (iv) identify whether the Gly82Ser variant is functionally important in altering sRAGE levels in an airway epithelial cell model.

Methods

Immunohistochemistry was used to identify RAGE protein expression in 26 human tissues and qPCR was used to quantify AGER mRNA in lung cells. Gene expression array data was used to identify AGER expression during lung development in 38 fetal lung samples. RNA-Seq was used to identify AGER transcripts in lung cells. sRAGE levels were assessed in cells and patient serum by ELISA. BEAS2B-R1 cells were transfected to overexpress RAGE protein with either the Gly82 or Ser82 variant and sRAGE levels identified.

Results

Immunohistochemical assessment of 6 adult lung samples identified high RAGE expression in the alveoli of healthy adults and individuals with COPD. AGER/RAGE expression increased across developmental stages in human fetal lung at both the mRNA (38 samples) and protein levels (20 samples). Extensive AGER splicing was identified. The rs2070600T (Ser82) allele is associated with higher FEV1, FEV1/FVC and lower serum sRAGE levels in UK smokers. Using an airway epithelium model overexpressing the Gly82 or Ser82 variants we found that HMGB1 activation of the RAGE-Ser82 receptor results in lower sRAGE production.

Conclusions

This study provides new information regarding the expression profile and potential role of RAGE in the human lung and shows a functional role of the Gly82Ser variant. These findings advance our understanding of the potential mechanisms underlying COPD particularly for carriers of this AGER polymorphism.

Citation

Miller, S., Henry, A. P., Hodge, E., Kheirallah, A. K., Billington, C. K., Rimington, T. L., Bhaker, S. K., Obeidat, M., Melén, E., Merid, S. K., Swan, C., Gowland, C., Nelson, C. P., Stewart, C. E., Bolton, C. E., Kilty, I., Malarstig, A., Parker, S. G., Moffatt, M. F., Wardlaw, A. J., …Sayers, I. (2016). The Ser82 RAGE Variant Affects Lung Function and Serum RAGE in Smokers and sRAGE Production In Vitro. PLoS ONE, 11(10), Article e0164041. https://doi.org/10.1371/journal.pone.0164041

• Journal Article Type: Article
• Acceptance Date: Sep 19, 2016
• Online Publication Date: Oct 18, 2016
• Publication Date: Oct 18, 2016
• Deposit Date: Nov 2, 2016
• Publicly Available Date: Nov 2, 2016
• Journal: PLOS ONE
• Electronic ISSN: 1932-6203
• Publisher: Public Library of Science
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 10
• Article Number: e0164041
• DOI: https://doi.org/10.1371/journal.pone.0164041
• Public URL: https://nottingham-repository.worktribe.com/output/823002
• Publisher URL: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0164041
• Contract Date: Nov 2, 2016