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Novel Roles for Notch3 and Notch4 Receptors in Gene Expression and Susceptibility to Ozone-Induced Lung Inflammation in Mice.

Verhein KC, McCaw Z, Gladwell W, Trivedi S, Bushel PR, Kleeberger SR - Environ. Health Perspect. (2015)

Bottom Line: We hypothesized that Notch3 and Notch4 are important determinants of susceptibility to ozone-induced lung inflammation.Relative to air-exposed controls, ozone increased bronchoalveolar lavage fluid (BALF) protein, a marker of lung permeability, in all genotypes, but significantly greater concentrations were found in Notch4-/- compared with WT and Notch3-/- mice.These novel findings are consistent with Notch3 and Notch4 as susceptibility genes for ozone-induced lung injury, and suggest that Notch receptors protect against innate immune inflammation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Respiratory Biology, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health (NIH), Department of Health and Human Resources (DHHS), Research Triangle Park, North Carolina, USA.

ABSTRACT

Background: Ozone is a highly toxic air pollutant and global health concern. Mechanisms of genetic susceptibility to ozone-induced lung inflammation are not completely understood. We hypothesized that Notch3 and Notch4 are important determinants of susceptibility to ozone-induced lung inflammation.

Methods: Wild-type (WT), Notch3 (Notch3-/-), and Notch4 (Notch4-/-) knockout mice were exposed to ozone (0.3 ppm) or filtered air for 6-72 hr.

Results: Relative to air-exposed controls, ozone increased bronchoalveolar lavage fluid (BALF) protein, a marker of lung permeability, in all genotypes, but significantly greater concentrations were found in Notch4-/- compared with WT and Notch3-/- mice. Significantly greater mean numbers of BALF neutrophils were found in Notch3-/- and Notch4-/- mice compared with WT mice after ozone exposure. Expression of whole lung Tnf was significantly increased after ozone in Notch3-/- and Notch4-/- mice, and was significantly greater in Notch3-/- compared with WT mice. Statistical analyses of the transcriptome identified differentially expressed gene networks between WT and knockout mice basally and after ozone, and included Trim30, a member of the inflammasome pathway, and Traf6, an inflammatory signaling member.

Conclusions: These novel findings are consistent with Notch3 and Notch4 as susceptibility genes for ozone-induced lung injury, and suggest that Notch receptors protect against innate immune inflammation.

No MeSH data available.


Related in: MedlinePlus

Gene expression microarray from whole-lung homogenates identified differentially expressed genes after exposure to ozone. KO, knockout. (A) In WT mice, 116 transcripts were differentially expressed at at least one time point after exposure to ozone compared with air. (B) In Notch3–/– mice, 739 transcripts were differentially expressed at at least one time point after ozone compared with air. (C) In Notch4–/– mice, 155 transcripts were differentially expressed at at least one time point after ozone compared with air. p < 0.05, ANOVA with Tukey’s HSD post hoc test and Benjamini-Hochberg multiple testing correction. (D) Number of significantly differentially expressed genes after ozone compared with air. (E) Top 10 significant biological functions from Ingenuity Pathway Analysis and numbers of differentially expressed genes in each category.
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f4: Gene expression microarray from whole-lung homogenates identified differentially expressed genes after exposure to ozone. KO, knockout. (A) In WT mice, 116 transcripts were differentially expressed at at least one time point after exposure to ozone compared with air. (B) In Notch3–/– mice, 739 transcripts were differentially expressed at at least one time point after ozone compared with air. (C) In Notch4–/– mice, 155 transcripts were differentially expressed at at least one time point after ozone compared with air. p < 0.05, ANOVA with Tukey’s HSD post hoc test and Benjamini-Hochberg multiple testing correction. (D) Number of significantly differentially expressed genes after ozone compared with air. (E) Top 10 significant biological functions from Ingenuity Pathway Analysis and numbers of differentially expressed genes in each category.

Mentions: Transcriptomic analysis identified candidate gene networks for Notch-mediated lung inflammation. Although TNF-α may influence a portion of Notch3-mediated protection against ozone-induced inflammation, additional mechanisms must contribute to the effect in mice with deletion of Notch3 and Notch4. To further understand the downstream pathways through which Notch3 and Notch4 mediate ozone-induced lung inflammation, we used genome-wide transcriptome analysis to identify candidate gene transcripts and interactomes. ANOVA modeling of the data identified few differentially expressed genes between WT and knockout mice after air exposure (see Supplemental Material, Table S2). However, ANOVA and pattern recognition EPIG (Chou et al. 2007) analyses identified ozone-induced changes in many gene transcripts in all genotypes (Figure 4A–C; see also Supplemental Material, Tables S2–S4).


Novel Roles for Notch3 and Notch4 Receptors in Gene Expression and Susceptibility to Ozone-Induced Lung Inflammation in Mice.

Verhein KC, McCaw Z, Gladwell W, Trivedi S, Bushel PR, Kleeberger SR - Environ. Health Perspect. (2015)

Gene expression microarray from whole-lung homogenates identified differentially expressed genes after exposure to ozone. KO, knockout. (A) In WT mice, 116 transcripts were differentially expressed at at least one time point after exposure to ozone compared with air. (B) In Notch3–/– mice, 739 transcripts were differentially expressed at at least one time point after ozone compared with air. (C) In Notch4–/– mice, 155 transcripts were differentially expressed at at least one time point after ozone compared with air. p < 0.05, ANOVA with Tukey’s HSD post hoc test and Benjamini-Hochberg multiple testing correction. (D) Number of significantly differentially expressed genes after ozone compared with air. (E) Top 10 significant biological functions from Ingenuity Pathway Analysis and numbers of differentially expressed genes in each category.
© Copyright Policy - public-domain
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4529014&req=5

f4: Gene expression microarray from whole-lung homogenates identified differentially expressed genes after exposure to ozone. KO, knockout. (A) In WT mice, 116 transcripts were differentially expressed at at least one time point after exposure to ozone compared with air. (B) In Notch3–/– mice, 739 transcripts were differentially expressed at at least one time point after ozone compared with air. (C) In Notch4–/– mice, 155 transcripts were differentially expressed at at least one time point after ozone compared with air. p < 0.05, ANOVA with Tukey’s HSD post hoc test and Benjamini-Hochberg multiple testing correction. (D) Number of significantly differentially expressed genes after ozone compared with air. (E) Top 10 significant biological functions from Ingenuity Pathway Analysis and numbers of differentially expressed genes in each category.
Mentions: Transcriptomic analysis identified candidate gene networks for Notch-mediated lung inflammation. Although TNF-α may influence a portion of Notch3-mediated protection against ozone-induced inflammation, additional mechanisms must contribute to the effect in mice with deletion of Notch3 and Notch4. To further understand the downstream pathways through which Notch3 and Notch4 mediate ozone-induced lung inflammation, we used genome-wide transcriptome analysis to identify candidate gene transcripts and interactomes. ANOVA modeling of the data identified few differentially expressed genes between WT and knockout mice after air exposure (see Supplemental Material, Table S2). However, ANOVA and pattern recognition EPIG (Chou et al. 2007) analyses identified ozone-induced changes in many gene transcripts in all genotypes (Figure 4A–C; see also Supplemental Material, Tables S2–S4).

Bottom Line: We hypothesized that Notch3 and Notch4 are important determinants of susceptibility to ozone-induced lung inflammation.Relative to air-exposed controls, ozone increased bronchoalveolar lavage fluid (BALF) protein, a marker of lung permeability, in all genotypes, but significantly greater concentrations were found in Notch4-/- compared with WT and Notch3-/- mice.These novel findings are consistent with Notch3 and Notch4 as susceptibility genes for ozone-induced lung injury, and suggest that Notch receptors protect against innate immune inflammation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Respiratory Biology, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health (NIH), Department of Health and Human Resources (DHHS), Research Triangle Park, North Carolina, USA.

ABSTRACT

Background: Ozone is a highly toxic air pollutant and global health concern. Mechanisms of genetic susceptibility to ozone-induced lung inflammation are not completely understood. We hypothesized that Notch3 and Notch4 are important determinants of susceptibility to ozone-induced lung inflammation.

Methods: Wild-type (WT), Notch3 (Notch3-/-), and Notch4 (Notch4-/-) knockout mice were exposed to ozone (0.3 ppm) or filtered air for 6-72 hr.

Results: Relative to air-exposed controls, ozone increased bronchoalveolar lavage fluid (BALF) protein, a marker of lung permeability, in all genotypes, but significantly greater concentrations were found in Notch4-/- compared with WT and Notch3-/- mice. Significantly greater mean numbers of BALF neutrophils were found in Notch3-/- and Notch4-/- mice compared with WT mice after ozone exposure. Expression of whole lung Tnf was significantly increased after ozone in Notch3-/- and Notch4-/- mice, and was significantly greater in Notch3-/- compared with WT mice. Statistical analyses of the transcriptome identified differentially expressed gene networks between WT and knockout mice basally and after ozone, and included Trim30, a member of the inflammasome pathway, and Traf6, an inflammatory signaling member.

Conclusions: These novel findings are consistent with Notch3 and Notch4 as susceptibility genes for ozone-induced lung injury, and suggest that Notch receptors protect against innate immune inflammation.

No MeSH data available.


Related in: MedlinePlus