GADD45a promoter regulation by a functional genetic variant associated with acute lung injury

Sumegha Mitra, Michael S. Wade, Xiaoguang Sun, Nurgul Moldobaeva, Carlos Flores, Ma Shwu-Fan, Wei Zhang, Joe G N Garcia, Jeffrey R. Jacobson

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Rationale: Growth arrest DNA damage inducible alpha (GADD45a) is a stress-induced gene we have shown to participate in the pathophysiology of ventilator-induced lung injury (VILI) via regulation of mechanical stress-induced Akt ubiquitination and phosphorylation. The regulation of GADD45a expression by mechanical stress and its relationship with acute lung injury (ALI) susceptibility and severity, however, remains unknown. Objectives: We examined mechanical stress-dependent regulatory elements (MSRE) in the GADD45a promoter and the contribution of promoter polymorphisms in GADD45a expression and ALI susceptibility. Methods and Results: Initial studies in GADD45a knockout and heterozygous mice confirmed the relationship of GADD45a gene dose to VILI severity. Human lung endothelial cells (EC) transfected with a luciferase vector containing the full length GADD45a promoter sequence (-771 to +223) demonstrated a >4 fold increase in GADD45a expression in response to 18% cyclic stretch (CS, 4 h) compared to static controls while specific promoter regions harboring CS-dependent MSRE were identified using vectors containing serial deletion constructs of the GADD45a promoter. In silico analyses of GADD45a promoter region (-371 to -133) revealed a potential binding site for specificity protein 1 (SP1), a finding supported by confirmed SP1 binding with the GADD45a promoter and by the significant attenuation of CS-dependent GADD45a promoter activity in response to SP1 silencing. Separately, case-control association studies revealed a significant association of a GADD45a promoter SNP at -589 (rs581000, G>C) with reduced ALI susceptibility. Subsequently, we found allelic variation of this SNP is associated with both differential GADD45a expression in mechanically stressed EC (18% CS, 4 h) and differential binding site of interferon regulatory factor 7 (IRF7) at this site. Conclusion: These results strongly support a functional role for GADD45a in ALI/VILI and identify a specific gene variant that confers risk for ALI.

Original languageEnglish (US)
Article numbere100169
JournalPLoS One
Volume9
Issue number6
DOIs
StatePublished - Jun 18 2014
Externally publishedYes

Fingerprint

Acute Lung Injury
Mechanical Stress
Ventilator-Induced Lung Injury
lungs
promoter regions
mechanical stress
Genes
Endothelial cells
ventilators
Genetic Promoter Regions
Single Nucleotide Polymorphism
Interferon Regulatory Factor-7
Endothelial Cells
Binding Sites
Association reactions
Phosphorylation
Proteins
Ubiquitination
endothelial cells
Polymorphism

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Mitra, S., Wade, M. S., Sun, X., Moldobaeva, N., Flores, C., Shwu-Fan, M., ... Jacobson, J. R. (2014). GADD45a promoter regulation by a functional genetic variant associated with acute lung injury. PLoS One, 9(6), [e100169]. https://doi.org/10.1371/journal.pone.0100169

GADD45a promoter regulation by a functional genetic variant associated with acute lung injury. / Mitra, Sumegha; Wade, Michael S.; Sun, Xiaoguang; Moldobaeva, Nurgul; Flores, Carlos; Shwu-Fan, Ma; Zhang, Wei; Garcia, Joe G N; Jacobson, Jeffrey R.

In: PLoS One, Vol. 9, No. 6, e100169, 18.06.2014.

Research output: Contribution to journalArticle

Mitra, S, Wade, MS, Sun, X, Moldobaeva, N, Flores, C, Shwu-Fan, M, Zhang, W, Garcia, JGN & Jacobson, JR 2014, 'GADD45a promoter regulation by a functional genetic variant associated with acute lung injury', PLoS One, vol. 9, no. 6, e100169. https://doi.org/10.1371/journal.pone.0100169
Mitra, Sumegha ; Wade, Michael S. ; Sun, Xiaoguang ; Moldobaeva, Nurgul ; Flores, Carlos ; Shwu-Fan, Ma ; Zhang, Wei ; Garcia, Joe G N ; Jacobson, Jeffrey R. / GADD45a promoter regulation by a functional genetic variant associated with acute lung injury. In: PLoS One. 2014 ; Vol. 9, No. 6.
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abstract = "Rationale: Growth arrest DNA damage inducible alpha (GADD45a) is a stress-induced gene we have shown to participate in the pathophysiology of ventilator-induced lung injury (VILI) via regulation of mechanical stress-induced Akt ubiquitination and phosphorylation. The regulation of GADD45a expression by mechanical stress and its relationship with acute lung injury (ALI) susceptibility and severity, however, remains unknown. Objectives: We examined mechanical stress-dependent regulatory elements (MSRE) in the GADD45a promoter and the contribution of promoter polymorphisms in GADD45a expression and ALI susceptibility. Methods and Results: Initial studies in GADD45a knockout and heterozygous mice confirmed the relationship of GADD45a gene dose to VILI severity. Human lung endothelial cells (EC) transfected with a luciferase vector containing the full length GADD45a promoter sequence (-771 to +223) demonstrated a >4 fold increase in GADD45a expression in response to 18{\%} cyclic stretch (CS, 4 h) compared to static controls while specific promoter regions harboring CS-dependent MSRE were identified using vectors containing serial deletion constructs of the GADD45a promoter. In silico analyses of GADD45a promoter region (-371 to -133) revealed a potential binding site for specificity protein 1 (SP1), a finding supported by confirmed SP1 binding with the GADD45a promoter and by the significant attenuation of CS-dependent GADD45a promoter activity in response to SP1 silencing. Separately, case-control association studies revealed a significant association of a GADD45a promoter SNP at -589 (rs581000, G>C) with reduced ALI susceptibility. Subsequently, we found allelic variation of this SNP is associated with both differential GADD45a expression in mechanically stressed EC (18{\%} CS, 4 h) and differential binding site of interferon regulatory factor 7 (IRF7) at this site. Conclusion: These results strongly support a functional role for GADD45a in ALI/VILI and identify a specific gene variant that confers risk for ALI.",
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