Glutaredoxin 1 controls monocyte reprogramming during nutrient stress and protects mice against obesity and atherosclerosis in a sex-specific manner

Figures

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  Fig. 1. Characterization of metabolic phenotypes of chow-fed aged male and female Grx1-/- mice as compared to age-matched male and female C57BL/6 J mice. a–b Body weights. c–d Fasting blood glucose levels. e–f Total plasma cholesterol levels and plasma lipoprotein profiles in female WT (n = 5) and Grx1-/-mice (n = 4) at 18 months of age. Lipoprotein profiles were generated from pooled plasma samples. All data are expressed as mean ± S.D., n = 5–6 mice per group for a–f; *One-way ANOVA followed by Fisher’s Least Significance Difference test was used to compare the mean values between experimental groups. P < 0.05; **P < 0.01; ***P < 0.001; Source data are provided as a Source Data file.

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  Fig. 2. Assessment of monocyte chemotactic activity in vivo and macrophage recruitment into gonadal adipose tissues from aged male and female mice. a Recruitment monocyte-derived macrophages into implanted Matrigel plugs loaded with. CP-1. b F4/80 mRNA expression in gonadal adipose tissues. c, d Representative images of gonadal adipose tissue section stained with H&E. Scale bar = 50 μm All data are expressed as mean ± S.D., n = 5–6 mice per group for a and b. One-way ANOVA followed by Fisher’s Least Significance Difference test was used to compare the mean values between experimental groups. *P < 0.05; **P < 0.01; ***P < 0.001. Source data are provided as a Source Data file.

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  Fig. 3. Grx1 deficiency in chow-fed aged female Grx1-/- mice promotes atherosclerotic lesion formation. a, b Images of Oil Red O-stained aorta from male and female 18-month-old Grx1-/- Mice and age-matched male and female C57BL/6 J mice fed a normal chow. c Quantitation of aortic lesion area. n = 5–6 mice per group. d Distribution of lesions between aortic arch, descending aorta and abdominal aorta in female Grx1-/- mice, n = 5 mice per group. All data are expressed as mean ± S.D. One-way ANOVA followed by Fisher’s Least Significance Difference test was used to compare the mean values between experimental groups. Source data are provided as a Source Data file.

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  Fig. 4. Grx1 deficiency in chow-fed female Grx1-/- mice promotes atherogenesis and drives macrophage recruitment and protein S-glutathionylation in atherosclerotic lesions. a Representative images of Oil Red O-stained sections from the aorta root of an 18-month-old female Grx1-/- mice and an age-matched male and female C57BL/6 J mouse (WT) maintained on a normal chow diet. Scale bar = 100 μm. Insert: Scale bar = 20 μm. b Quantitation of lesion area in the aortic root of male and female Grx1-/- mice and age-matched male and female C57BL/6 J mice (WT) maintained on a normal chow diet for 18 months. n = 5–6 mice per group. c Representative images of aorta root sections stained with either rat IgG2a (Control) or a macrophage-specific antiserum against CD68 (CD68) from an 18-month-old female Grx1-/- mice and an age-matched female C57BL/6 J mouse (WT). Scale bar = 50 μm. d Macrophage content of atherosclerotic lesions in the aorta root of 18-month-old female Grx1-/- mice and age-matched male and female C57BL/6 J mice (WT) maintained on a normal chow diet for 18 months. n = 5–6 mice per group with male and female Grx1-/- mice and age-matched male and female C57BL/6 J mice (WT). e,f…

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  Fig. 5. Macrophage Reprogramming in Aged Grx1-Deficient Mice. a Heatmap representing normalized gene expression in peritoneal macrophages isolated from male and female Grx1-/- mice and age-matched male and female C57BL/6 J mice (WT) maintained on a normal chow diet for 18 months. b Principal component analysis (PCA) of variances in the mRNA expression in peritoneal macrophages of male (blue) and female (red) WT and Grx1-/- mice (KO). c Venn diagram displaying differentially expressed genes between peritoneal macrophages from male (blue) and female mice (red). d Derepession of NOX4 expression by Grx1 deficiency in peritoneal macrophages from 18-month-old female but not male mice. e Derepession of NOS3 expression by Grx1 deficiency in peritoneal macrophages from 18-month-old female but not male mice. All data are expressed as mean ± S.D, n = 3. One-way ANOVA followed by Fisher’s Least Significance Difference test was used to compare the mean values between experimental groups. Source data are provided as a Source Data file.

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  Fig. 6. Hematopoietic Grx1 deficiency promotes formation of nitrotyrosine in the aortic root of aged female but not male mice. a Representative images of aorta root sections stained with an antiserum against nitrotyrosine from 18-month-old male and female WT and Grx1-/- mice maintained on a normal chow diet for 18 months. Scale bar: 500 μm. Inserts: 100x magnification. b Nitrotyrosine levels measured in the aortic root of male and female WT and Grx1-/- mice maintained on a normal chow diet for 18 months. All data are expressed as mean ± S.D, n = 5–6 per group, 8 aortic root sections per mouse. Source data are provided as a Source Data file.

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  Fig. 7. Hematopoietic Grx1 deficiency accelerates body weight gain in reproductively healthy female LDLR-/- mice fed a high-calorie diet. a Body weights. n = 14 per group. b Fasting blood glucose levels. n = 14 per group. c Total cholesterol levels. n = 14 per group. d Total plasma triglyceride levels. n = 14 per group. e Lipoprotein profiles of LDLR-/- (WT) and LDLR-/-Grx1Leuko-/- mice (Grx1Leuko-/-) from pooled plasma samples; n = 14 per group. All data are expressed as mean ± S.E. One-way ANOVA followed by Fisher’s Least Significance Difference test was used to compare the mean values between experimental groups. Source data are provided as a Source Data file.

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  Fig. 8. Hematopoietic Grx1 deficiency promotes atherosclerosis in reproductively healthy female LDLR-/- mice by enhancing recruitment of monocyte-derived macrophages into atherosclerotic lesions. a Representative images of Oil Red O-stained aortas from female LDLR-/-Grx1Leuko-/- mice (Grx1Leuko-/-) and age-matched female LDLR-/- mice (WT) fed a HCD. b Quantification of aortic lesion area. n = 14 per group. Week 6: n = 10; Week 20: n = 14. c Representative images of Oil Red O-stained sections from the aorta roots of HCD-fed female LDLR-/-Grx1Leuko-/- (Grx1Leuko-/-) and age-matched female LDLR-/- mice (WT). Scale bar = 500 μm. d Quantitation of lesion area in the aortic root. Week 6: n = 10; Week 20: n = 14 per group. e Distribution of lesion size across a 500 μm segment of the aortic root beginning at the aortic valve. n = 14 mice per group. f Recruitment monocyte-derived macrophages into implanted Matrigel plugs loaded with. MCP-1. n = 14 mice per group. g Representative images of aortic root sections stained with a macrophage-specific antiserum against CD68. Scale bar = 500 μm. h Quantitation of macrophage content in atherosclerotic lesions in the aortic root. Week 6: n = 10; Week 20: n…

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  Fig. 9. Hematopoietic Grx1 deficiency exacerbates nutrient stress-induced monocyte dysfunction. a MKP-1 activity in purified peritoneal macrophages isolated from HCD-fed female LDLR-/-Grx1Leuko-/- mice (Grx1Leuko-/-) and age-matched female LDLR-/- mice (WT). n = 15 mice per group. b Chemotaxis of purified peritoneal macrophages isolated from female Grx1Leuko-/- and age-matched C57BL/6 mice (WT). n = 4 mice per group. All data are expressed as mean ± S.D. One-way ANOVA followed by Fisher’s Least Significance Difference test were used to compare the mean values between experimental groups. Source data are provided as a Source Data file.

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  Fig. 10. Macrophage-restricted overexpression of Grx1 in LDLR-/- mice reverses the sexually dimorphic expression of NOX, NOS, and ARG2 genes induced by HCD feeding. a Grx1 mRNA expression in bone marrow-derived macrophages isolated from 30-week-old male and female maintenance diet (MD)-fed LDLR-/- mice (gray circles, n = 3) and HCD-fed LDLR-/- mice overexpressing either EGFP (red circles) or Grx1 (green circles), n = 4–6 mice per group. b MKP-1 activity in bone marrow-derived macrophages isolated from 30-week-old male and female maintenance diet (MD)-LDLR-/- mice (gray circles, n = 3) and HCD-fed LDLR-/- mice overexpressing either EGFP (red circles) or Grx1 (green circles), n = 4–6 mice per group. c–i Gene expression profiling of NOX1, 2 and 4, ARG2 and NOS1, 2 and 3 in bone marrow-derived macrophages isolated from 30-week-old male and female maintenance diet (MD)-LDLR-/- mice (gray circles, n = 3) and HCD-fed LDLR-/- mice overexpressing either EGFP (red circles) or Grx1 (green circles), n = 4–6 mice per group. (j) Nitrotyrosine levels measured in the aortic root of 30-week-old male-female maintenance diet (MD)-LDLR-/- mice (gray circles, n = 3) and HCD-fed LDLR-/- mice overexpressing…