Supplementary data from: Systems genomics reveals age- and sex-dependent metabolic dysregulation from Glo1 reduction in mice
Data files
Feb 06, 2026 version files 1.17 MB
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README.md
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Supplementary_Figure_1.pdf
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Supplementary_Figure_2.pdf
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Supplementary_Figure_3.pdf
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Supplementary_Figure_4.pdf
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Supplementary_Figure_5.pdf
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Supplementary_Figure_6.pdf
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Supplementary_Figure_7.pdf
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Supplementary_Table_1.pdf
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Supplementary_Table_2.pdf
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Abstract
Objectives: Glyoxalase 1 (Glo1) detoxifies reactive dicarbonyl compounds such as methylglyoxal, a precursor of advanced glycation end products (AGEs), which contribute to metabolic disorders. However, the contribution of AGE-independent mechanisms to Glo1-related metabolic dysfunction remains unclear.
Methods: We conducted a longitudinal study in male and female Glo1 heterozygous knockdown (Glo1⁺/⁻) mice (~50% Glo1 expression). Metabolic phenotypes, including body weight, adiposity, glycemic control, and plasma lipid levels, were assessed over time. Atherosclerotic burden, AGE levels, and gene expression profiles in liver, adipose, muscle, kidney, and aorta were examined to identify pathway alterations and regulatory genes affected by Glo1 reduction.
Results: Partial Glo1 loss resulted in obesity, hyperglycemia, dyslipidemia, and altered lipid metabolism in an age- and sex-dependent manner, with most phenotypes emerging after ~14 weeks. Glo1⁺/⁻ females exhibited impaired glycemic control and elevated triglycerides, along with perturbations in adipogenesis, PPARγ signaling, insulin signaling, and fatty acid metabolism in liver and adipose tissue. Glo1⁺/⁻ males displayed increased skeletal muscle mass and visceral adiposity with changes in lipid metabolic pathways. Methylglyoxal-derived AGE accumulation was altered only in male skeletal muscle and did not explain broader phenotypes. Transcriptomic analyses suggest altered glucose and lipid metabolism may be partially driven by alternative detoxification of methylglyoxal to metabolites such as pyruvate. Transcription factor analysis identified Hnf4a (across tissues) and Arntl (in aorta, liver, and kidney) as female-biased regulators altered by Glo1 deficiency.
Conclusions: Glo1 reduction disrupts metabolic health through sex- and age-dependent pathways largely independent of AGE accumulation, involving tissue-specific metabolic reprogramming and transcriptional regulation.
Dataset DOI: 10.5061/dryad.83bk3jb6x
Description of the data and file structure
The data were collected from a mouse model of reduced glyoxalase 1 activity using heterozygous Glo1⁺/⁻ knockdown mice and littermate controls. Animals were genotyped by PCR and followed longitudinally from weaning into adulthood with repeated metabolic phenotyping, including weekly body weight, biweekly NMR-based body composition, and serial intraperitoneal glucose tolerance tests; fasting plasma was collected at multiple ages for lipid, glucose, and insulin measurements. At study endpoints, key metabolic tissues (e.g., liver, kidney, gonadal adipose, skeletal muscle, and aorta) were harvested for Glo1 enzyme activity assays, AGE quantification by ELISA (MGH1 and CEL), protein abundance by Western blot (e.g., Glo1 and RAGE), and targeted gene expression by qPCR. In parallel, atherosclerotic burden was assessed histologically by Oil Red O staining of aortic sinus sections, and a multi-tissue transcriptomic microarray dataset (Illumina MouseRef-8 v2.0) was generated in 34-week-old female mice to identify differentially expressed genes for downstream pathway and integrative genomics analyses.
- Supplementary_Table_1.pdf – qPCR primer sequences used in mouse genotyping and tissue gene-expression assays.
Legend: Supplementary Table 1. List of qPCR mouse primers (forward and reverse) for Glo1/Gapdh and metabolic/AGE-related genes assayed by SYBR Green qPCR.
- Supplementary_Table_2.pdf – Results table for cross-species overlap analyses (mouse Glo1-regulated genes vs human sex-/age-biased programs).
Legend: Supplementary Table 2. Overlap analysis between tissue-specific Glo1 differential expression signatures in mice and human sex-biased and age-associated gene sets from GTEx and GenAge. The table reports overlapping gene symbols, counts, Fisher’s exact test p-values, odds ratios, and background/comparator set sizes.
- Supplementary_Figure_1.pdf – Validation of the Glo1 knockdown (Glo1⁺/⁻) mouse model at the DNA and protein levels.
Legend: Supplementary Figure 1. (A) PCR-based genotyping showing the internal control (Gapdh; top band) and the Glo1 knockdown-specific band (~220 bp; bottom band), with WT lacking the KD band. (B) Representative immunoblots showing Glo1 protein levels in male and female WT vs Glo1 KD mice with actin as a loading control (examples of KD/WT shown).
- Supplementary_Figure_2.pdf – Longitudinal monitoring of food and water intake by sex and genotype.
Legend: Supplementary Figure 2. Weekly chow and water intake measurements in WT and Glo1 KD mice across development. (A–B) Female weekly chow (g) and water (mL) intake. (C–D) Male weekly chow (g) and water (mL) intake. Intake was recorded weekly to track caloric/fluid consumption across experimental groups.
- Supplementary_Figure_3.pdf – Glucose tolerance testing across ages in male mice.
Legend: Supplementary Figure 3. Intraperitoneal glucose tolerance tests (IPGTT) in male WT vs Glo1 KD mice at multiple ages. (A) IPGTT area under the curve (AUC) at 5, 12, 23, and 33 weeks. (B–E) Glucose curves (0–120 min) following IP glucose challenge at the indicated ages, with reported p-values for genotype, time, and genotype×time effects.
- Supplementary_Figure_4.pdf – Fasting plasma glucose and insulin across ages, separated by sex.
Legend: Supplementary Figure 4. Fasting plasma glucose and insulin concentrations in WT and Glo1 KD mice at 7, 12, and 28 weeks. (A–B) Female glucose (mg/dL) and insulin (pg/mL). (C–D) Male glucose (mg/dL) and insulin (pg/mL). Measurements were obtained from overnight-fasted animals.
- Supplementary_Figure_5.pdf – Additional fasting lipid measures across ages, separated by sex.
Legend: Supplementary Figure 5. Fasting plasma lipid measurements in WT and Glo1 KD mice at 7, 12, and 28 weeks. (A–C) Female unesterified cholesterol (UC), free fatty acids (FFA), and LDL cholesterol. (D–F) Male UC, FFA, and LDL cholesterol.
- Supplementary_Figure_6.pdf – AGE/RAGE inflammatory signaling schematic and liver pathway-level expression changes.
Legend: Supplementary Figure 6. (A) Schematic of AGE–RAGE signaling leading to ROS activation, NFKB signaling, and downstream inflammatory/apoptotic programs. (B) Heatmaps of liver gene expression changes (WT vs Glo1 KD) across functional categories, including ROS/NFKB signaling, apoptosis regulators, cell-cycle regulators, cytokines, chemokines, and adhesion molecules.
- Supplementary_Figure_7.pdf – Differential expression of key glucose metabolism enzymes in adipose and aorta.
Legend: Supplementary Figure 7. Heatmap of log2 fold-change (Glo1 KD vs WT) for selected glycolysis/gluconeogenesis enzymes in adipose and aorta. Asterisks denote statistical significance (* p<0.05, ** p<0.01, *** p<0.001).