Gene dosage imbalance disrupts systemic metabolism in the Dp16 Down syndrome mouse model
- Department of Physiology, Pharmacology and Therapeutics, Johns Hopkins University, School of Medicine, United States
- Center for Metabolism and Obesity Research, Johns Hopkins University, School of Medicine, United States
- Department of Biological Chemistry, University of California, Irvine, United States
- Center for Epigenetics and Metabolism, University of California Irvine, United States
- The Children's Guild Foundation Down Syndrome Research Program, Department of Cancer Genetics and Genomics, Roswell Park Comprehensive Cancer Center, United States
- Genetics, Genomics and Bioinformatics Program, State University of New York at Buffalo, United States
- Department of Neuroscience, Johns Hopkins University School of Medicine, United States
Figures
Figure 1
Increased gene expression dosage of the triplicated Hsa21 gene orthologs on mouse chromosome 16 (Mmu16) across tissues.
(A) Graphical representation of human chromosome 21 (Hsa21) and the syntenic Mmu16 segment that is duplicated in Dp16 mice. (B) Global view of the expression of 108 triplicated Hsa21 gene orthologs on Mmu16 in gonadal white adipose tissue (gWAT), inguinal white adipose tissue (iWAT), interscapular brown adipose tissue (BAT), skeletal muscle (gastrocnemius), and hypothalamus. Red denotes transcript that is expressed at >1.5-fold the WT level, whereas blue denotes transcript that is expressed at significantly lower level compared to WT control. The Ktrap gene cluster (23 Ktrap genes) located between Cldn8 and Tiam1 is not shown. (C) Overlap analysis showing differentially expressed Hsa21 gene orthologs that are shared between males and females across six tissues. The criteria for differentially expressed genes (DEGs) is log2(FC)>0 with padj <0.05. n=6 RNA samples per genotype per sex per tissue-type. Chow-fed WT and Dp16 mice were at 27.5 weeks of age at the time of tissue collection.
Figure 2 with 4 supplements
Sexual dimorphism in body weight, body temperature, food intake, and physical activity in chow-fed Dp16 mice.
(A) Body weight of chow-fed male Dp16 and WT mice over time. (B) Absolute and relative (% of body weight) fat and lean mass in male mice (WT = 15; Dp16 = 12). (C) Body weight of chow-fed female Dp16 and WT mice over time. (D) Absolute and relative (% of body weight) fat and lean mass in female mice (WT = 15; Dp16 = 15). (E–F) Food intake, total physical activity level, and energy expenditure of male (E) and female (F) Dp16 and WT mice across the circadian cycle (light and dark) and metabolic states (ad libitum fed, fast, refeed). Sample size for male (WT = 10–12; Dp16 = 11–12) and female (WT = 13–15; Dp16 = 5–6) mice. (G–H) Fecal frequency, average fecal weight, and fecal energy content (per gram and total) in male (G) and female (H) Dp16 and WT mice. Sample size for male (WT = 6; Dp16 = 6) and female (WT = 6; Dp16 = 6) mice. (I–J) Body temperature in the light and dark cycle of male (I) and female (J) Dp16 and WT mice. Sample size for male (WT = 10; Dp16 = 10) and female (WT = 15; Dp16 = 15) mice. All data are presented as mean ± SEM. * p<0.05; *** p<0.001; **** p<0.0001. For body weight over time, data were analyzed by 2-way ANOVA with Sidek post hoc tests.
Figure 2—figure supplement 1
Body and tissue weights of chow-fed male and female mice at termination of study.
Tissues were collected from chow-fed male and female mice at 27.5 weeks of age. Body weights and the absolute (A and C) and relative (B and D; % of body weight) weights of gWAT, iWAT, liver, and kidney in Dp16 and WT male (A–B) and female (C–D) mice. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue. Sample size: WT male = 10; Dp16 male = 30; WT female = 15; Dp16 female = 10. All data are presented as mean ± SEM. * p<0.05; ** p<0.01; *** p<0.001.
Figure 2—figure supplement 2
ANCOVA analysis of energy expenditure in chow-fed mice where lean mass is used as a covariate.
ANCOVA analysis of WT and Dp16 male mice across the circadian cycle (dark and light) in ad libitum fed (A), fasted (B), and refed (C) states. ANCOVA analysis of WT and Dp16 female mice across the circadian cycle (dark and light) in ad libitum fed (D), fasted (E), and refed (F) states. Male Sample size: WT = 12; Dp16 = 12. Female sample size: WT = 15; Dp16 = 6.
Figure 2—figure supplement 3
Reduced mitochondrial activity in the brown adipose tissue (BAT) of Dp16 mice.
Mitochondrial respiration through complex I (CI), CII, and CIV in BAT of WT and Dp16 male and female mice fed a standard chow. (A and D) Average oxygen consumption rate (OCR) traces per group, normalized to mitochondrial content. Each group tracing represents the average trace of 10 WT and 9–10 Dp16 samples. Each tracing shows the entire process of the Seahorse-based respirometry assay with injection compounds listed at the time of introduction to the sample. The sequence is as follows: (i) basal reads, (ii) addition of NADH (activation of respiration through complex I), (iii) addition of antimycin A (AA, inhibitor of complex III) and rotenone (Rot, inhibitor of complex I), (iv) addition of TMPD and ascorbate (to activate complex IV via electron donation to cytochrome c), and finally (v) addition of azide (inhibitor of complex IV). (B and E) The same information as presented in (A and D) conducted on the same samples, but the NADH injection step is replaced with the injection of succinate (to activate respiration through complex II) and rotenone (to inhibit complex I). (C and F) Average values of all data presented for BAT. Each data point represents the average of three technical replicates measured at three separate times. Both independent measurements of complex IV (CIV) were used to determine average CIV respiration. **** p<0.0001 (two-way ANOVA with Sidak’s multiple comparison).
Figure 2—figure supplement 4
Serum Triiodothyronine (T3), sex, and stress hormone levels in WT and Dp16 mice fed a standard chow.
(A) Serum T3 levels in male and female mice. (B) Serum testosterone levels in male mice. (C) Serum estradiol levels in female mice. (D) Serum corticosterone in male and female mice. Sample size: male WT = 8–10; male Dp16 = 25–30; female WT = 14–15; female Dp16 = 10.
Figure 3 with 2 supplements
Glucose intolerance, insulin resistance, and impaired lipid clearance in chow-fed Dp16 mice.
(A–B) Overnight fasting insulin, blood glucose, serum triglyceride, cholesterol, non-esterified free fatty acids (NEFA), and β-hydroxybutyrate (ketone) in male (A) and female (B) Dp16 and WT mice. Sample size for male mice (WT = 15; Dp16 = 12) and female mice (WT = 14; Dp16 = 15). (C–F) Impaired glucose tolerance as determined by the glucose tolerance test (GTT) in male (C) and female (E) Dp16 mice compared to WT controls. Impaired insulin sensitivity as determined by the insulin tolerance test (ITT) in male (D) and female (F) Dp16 compared to WT controls. Sample size for male mice (WT = 15; Dp16 = 12) and female mice (WT = 14; Dp16 = 15). (G–H) Impaired triglyceride clearance in response to lipid gavage as determined by the lipid tolerance test (LTT) in male (G) and female (H) Dp16 relative to WT controls. Sample size for male mice (WT = 10; Dp16 = 14) and female mice (WT = 15; Dp16 = 15). (I–J) Pooled mouse sera from male (I) and female (J) Dp16 and WT mice were fractionated by fast protein liquid chromatography (FPLC), and the triglyceride and cholesterol content of each fraction was quantified. Fractions corresponding to very-low density lipoprotein (VLDL), low-density lipoprotein (LDL), intermediate-density lipoprotein (IDL), and high-density lipoprotein (HDL) are indicated. All data are presented as mean ± SEM. * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001. For all tolerance tests, data were analyzed by two-way ANOVA with Sidek post hoc tests.
Figure 3—figure supplement 1
Liver triacylglycerol (TAG), diacylglycerol (DAG), and cholesterol levels in chow-fed Dp16 mice.
Quantification of hepatic TAG and DAG (by TLC method), and cholesterol (by infinity assay kit) levels in chow-fed Dp16 male (A–C) and female mice (D–F) and their corresponding WT controls. Sample size: male WT = 10 and Dp16 = 30; female WT = 15 and Dp16 = 10.
Figure 3—figure supplement 2
Mitochondrial activity in the liver of Dp16 mice.
Mitochondrial respiration through complex I (CI), CII, and CIV in the liver of WT and Dp16 male and female mice fed a standard chow. (A and D) Average oxygen consumption rate (OCR) traces per group, normalized to mitochondrial content. Each group tracing represents the average trace of 10 WT and 10 Dp16 samples. Each tracing shows the entire process of the Seahorse-based respirometry assay with injection compounds listed at the time of introduction to the sample. The sequence is as follows: (i) basal reads, (ii) addition of NADH (activation of respiration through complex I), (iii) addition of antimycin A (AA, inhibitor of complex III) and rotenone (Rot, inhibitor of complex I), (iv) addition of TMPD and ascorbate (to activate complex IV via electron donation to cytochrome c), and finally (v) addition of azide (inhibitor of complex IV). (B and E) The same information as presented in (A and D) conducted on the same samples, but the NADH injection step is replaced with the injection of succinate (to activate respiration through complex II) and rotenone (to inhibit complex I). (C and F) Average values of all data presented for liver. Each data point represents the average of three technical replicates measured at three separate times. Both independent measurements of complex IV (CIV) were used to determine average CIV respiration.
Figure 4 with 4 supplements
Altered liver and serum metabolome in Dp16 male and female mice.
(A–B) Partial least squares discrimination analysis (PLS-DA) of liver and serum metabolites of Dp16 and WT males and females. N=6 samples per genotype per sex. (C) Venn diagram of differential metabolites shared between liver and serum in Dp16 male or female mice. (D) Venn diagram of differential liver or serum metabolites shared between Dp16 males and females. (E) KEGG enrichment showing altered metabolic processes in Dp16 female serum. ES, enrichment score; NES, normalized enrichment score.
-
Figure 4—source data 1
Differential metabolites in Dp16 male mouse liver vs WT controls.
Differential metabolites criteria: VIP >1.0, fold change (FC)>1.2 or FC <0.833 and p-value <0.05. Sample name notation: male WT liver (M_WT_L), male WT serum (M_WT_S), male Dp16 liver (M_16_L), male Dp16 serum (M_16_S), female WT liver (F_WT_L), female WT serum (F_WT_L), female Dp16 liver (F_16_L), female Dp16 serum (F_16_S).
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig4-data1-v1.xlsx
-
Figure 4—source data 2
Differential metabolites in Dp16 female mouse liver vs WT controls.
Differential metabolites criteria: VIP >1.0, fold change (FC)>1.2 or FC <0.833 and p-value <0.05. Sample name notation: male WT liver (M_WT_L), male WT serum (M_WT_S), male Dp16 liver (M_16_L), male Dp16 serum (M_16_S), female WT liver (F_WT_L), female WT serum (F_WT_L), female Dp16 liver (F_16_L), female Dp16 serum (F_16_S).
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig4-data2-v1.xlsx
-
Figure 4—source data 3
Differential metabolites in Dp16 male mouse serum vs WT controls.
Differential metabolites criteria: VIP >1.0, fold change (FC)>1.2 or FC <0.833 and <i>P-value <0.05. Sample name notation: male WT liver (M_WT_L), male WT serum (M_WT_S), male Dp16 liver (M_16_L), male Dp16 serum (M_16_S), female WT liver (F_WT_L), female WT serum (F_WT_L), female Dp16 liver (F_16_L), female Dp16 serum (F_16_S).
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig4-data3-v1.xlsx
-
Figure 4—source data 4
Differentially expressed metabolites in Dp16 female mouse serum vs WT controls.
Differential metabolites criteria: VIP >1.0, fold change (FC)>1.2 or FC <0.833 and <i>P-value <0.05. Sample name notation: male WT liver (M_WT_L), male WT serum (M_WT_S), male Dp16 liver (M_16_L), male Dp16 serum (M_16_S), female WT liver (F_WT_L), female WT serum (F_WT_L), female Dp16 liver (F_16_L), female Dp16 serum (F_16_S).
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig4-data4-v1.xlsx
-
Figure 4—source data 5
Shared and distinct differential metabolites in Dp16 male and female mouse liver and serum vs WT controls.
Sample name notation: male WT liver (M_WT_L), male WT serum (M_WT_S), male Dp16 liver (M_16_L), male Dp16 serum (M_16_S), female WT liver (F_WT_L), female WT serum (F_WT_L), female Dp16 liver (F_16_L), female Dp16 serum (F_16_S).
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig4-data5-v1.xlsx
Figure 4—figure supplement 1
Differential metabolites found in the liver and serum of Dp16 male and female mice.
Volcano plots showing differential metabolites up- and down-regulated in Dp16 male liver (A), female liver (B), male serum (C), and female serum (D). n=6 per genotype. VIP, Variable Importance in Projection. VIP scores provide a quantitative measure of a metabolite’s discriminatory power between different groups. Metabolites with a VIP score of 1.0 or greater are considered significant.
Figure 4—figure supplement 2
KEGG classification analysis of liver metabolites.
KEGG classification plots based on the differential metabolites from male (A) and female (B) Dp16 mouse liver vs WT control. The horizontal coordinates in the graph indicate the number of metabolites annotated under a particular KEGG pathway as a percentage of the number of all annotated metabolites, the vertical coordinates are KEGG pathway primary classifications on the right and KEGG pathway secondary classifications on the left.
Figure 4—figure supplement 3
KEGG classification analysis of serum metabolites.
KEGG classification plots based on the differential metabolites from male (A) and female (B) Dp16 mouse liver vs WT control. The horizontal coordinates in the graph indicate the number of metabolites annotated under a particular KEGG pathway as a percentage of the number of all annotated metabolites, the vertical coordinates are KEGG pathway primary classifications on the right and KEGG pathway secondary classifications on the left.
Figure 4—figure supplement 4
Serum alanine aminotransferase (ALT) levels in WT and Dp16 mice fed a standard chow.
Serum ALT levels in male and female mice. Sample size: male WT = v10; male Dp16=27; female WT = v14; female Dp16=10.
Figure 5 with 6 supplements
Transcriptomic changes and altered biological pathways across tissues in chow-fed Dp16 male and female mice.
(A) Number of differentially expressed genes (DEGs) that up or down regulated across six tissues in male and female Dp16 mice and their WT littermate controls. DEG is defined as any gene with log2(FC)>0.5 and padj <0.05. N=6 per genotype per tissue. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue; BAT, brown adipose tissue. (B) Overlap analysis showing DEGs that are shared between males and females, as well as those DEGs found in males or females only, across six tissues. (C) Gene ontology highlighting some of the top biological pathways altered across six tissues in male and female Dp16 mice.
-
Figure 5—source data 1
Differentially expressed genes (DEGs) upregulated in the gonadal white adipose tissue (gWAT) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data1-v1.xls
-
Figure 5—source data 2
Differentially expressed genes (DEGs) down-regulated in the gonadal white adipose tissue (gWAT) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data2-v1.xls
-
Figure 5—source data 3
Differentially expressed genes (DEGs) upregulated in the inguinal white adipose tissue (iWAT) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data3-v1.xls
-
Figure 5—source data 4
Differentially expressed genes (DEGs) down-regulated in the inguinal white adipose tissue (iWAT) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data4-v1.xls
-
Figure 5—source data 5
Differentially expressed genes (DEGs) upregulated in the brown adipose tissue (BAT) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data5-v1.xls
-
Figure 5—source data 6
Differentially expressed genes (DEGs) down-regulated in the brown adipose tissue (BAT) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data6-v1.xls
-
Figure 5—source data 7
Differentially expressed genes (DEGs) upregulated in the liver of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data7-v1.xls
-
Figure 5—source data 8
Differentially expressed genes (DEGs) down-regulated in the liver of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data8-v1.xls
-
Figure 5—source data 9
Differentially expressed genes (DEGs) upregulated in the skeletal muscle (gastrocnemius) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data9-v1.xls
-
Figure 5—source data 10
Differentially expressed genes (DEGs) down-regulated in the skeletal muscle (gastrocnemius) of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data10-v1.xls
-
Figure 5—source data 11
Differentially expressed genes (DEGs) upregulated in the hypothalamus of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data11-v1.xls
-
Figure 5—source data 12
Differentially expressed genes (DEGs) down-regulated in the hypothalamus of chow-fed Dp16 male mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data12-v1.xls
-
Figure 5—source data 13
Differentially expressed genes (DEGs) upregulated in the gonadal white adipose tissue (gWAT) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data13-v1.xls
-
Figure 5—source data 14
Differentially expressed genes (DEGs) down-regulated in the gonadal white adipose tissue (gWAT) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data14-v1.xls
-
Figure 5—source data 15
Differentially expressed genes (DEGs) upregulated in the inguinal white adipose tissue (iWAT) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data15-v1.xls
-
Figure 5—source data 16
Differentially expressed genes (DEGs) down-regulated in the inguinal white adipose tissue (iWAT) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data16-v1.xls
-
Figure 5—source data 17
Differentially expressed genes (DEGs) upregulated in the brown adipose tissue (BAT) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data17-v1.xls
-
Figure 5—source data 18
Differentially expressed genes (DEGs) down-regulated in the brown adipose tissue (BAT) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data18-v1.xls
-
Figure 5—source data 19
Differentially expressed genes (DEGs) upregulated in the liver of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data19-v1.xls
-
Figure 5—source data 20
Differentially expressed genes (DEGs) down-regulated in the liver of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data20-v1.xls
-
Figure 5—source data 21
Differentially expressed genes (DEGs) upregulated in the skeletal muscle (gastrocnemius) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data21-v1.xls
-
Figure 5—source data 22
Differentially expressed genes (DEGs) down-regulated in the skeletal muscle (gastrocnemius) of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data22-v1.xls
-
Figure 5—source data 23
Differentially expressed genes (DEGs) upregulated in the hypothalamus of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data23-v1.xls
-
Figure 5—source data 24
Differentially expressed genes (DEGs) down-regulated in the hypothalamus of chow-fed Dp16 female mice relative to WT controls.
- https://cdn.elifesciences.org/articles/110476/elife-110476-fig5-data24-v1.xls
Figure 5—figure supplement 1
Differentially expressed genes (DEGs) involved in ER stress, fibrosis, glucose and lipid metabolism that are up- or down-regulated in the inguinal white adipose tissue (iWAT) of Dp16 mice.
Figure 5—figure supplement 2
Differentially expressed genes (DEGs) involved in immune activation, lipid metabolism, and mitochondrial respiration that are up- or down-regulated in the brown adipose tissue (BAT) of Dp16 mice.
Figure 5—figure supplement 3
Differentially expressed genes (DEGs) involved in immune activation, lipid metabolism, and mitochondrial respiration that are up- or down-regulated in the liver of Dp16 mice.
Figure 5—figure supplement 4
Differentially expressed genes (DEGs) involved in immune response, metabolism, mitochondrial respiration, and Wnt signaling that are up- or down-regulated in the skeletal muscle (gastrocnemius) of Dp16 mice.
Figure 5—figure supplement 5
Differentially expressed genes (DEGs) involved in immune response and extracellular matrix that are upregulated in the hypothalamus of Dp16 mice.
Figure 5—figure supplement 6
Hydroxyproline (marker of fibrosis) and malondialdehyde (marker of oxidative stress) levels in the liver, gWAT, and iWAT of chow-fed Dp16 mice.
(A–F) Quantification of hydroxyproline content in the liver, gWAT, and iWAT of Dp16 male (A–C) and female (D–F) mice and their corresponding WT controls. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue. Sample size: male WT = 7–10 and Dp16 = 27–29; female WT = 10–13 and Dp16 = 7–10. (G–L) Quantification of malondialdehyde (MDA) levels in the liver, gWAT, and iWAT of Dp16 male (G–I) and female (J–L) mice and their corresponding WT controls. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue. Sample size: male WT = 6–10 and Dp16 = 25–30; female WT = 8–13 and Dp16 = 6–10. All data are presented as mean ± SEM. * P<0.05; ** P<0.01.
Figure 6 with 4 supplements
Sexually dimorphism in body weight, body temperature, food intake, and physical activity in Dp16 mice in response to a high-fat diet (HFD).
(A) Body weight of HFD-fed male Dp16 and WT mice over time. (B) Absolute and relative (% of body weight) fat and lean mass in male mice (WT = 15; Dp16 = 12). (C) Body weight of HFD-fed female Dp16 and WT mice over time. (D) Absolute and relative (% of body weight) fat and lean mass in female mice (WT = 14; Dp16 = 14). (E–F) Food intake, total physical activity level, and energy expenditure of male (E) and female (F) Dp16 and WT mice across the circadian cycle (light and dark) and metabolic states (ad libitum fed, fast, refeed). Sample size for male (WT = 8; Dp16 = 11) and female (WT = 12; Dp16 = 12) mice. (G–H) Fecal frequency, average fecal weight, and fecal energy content (per gram and total) in male (G) and female (H) Dp16 and WT mice on HFD. Sample size for male (WT = 6; Dp16 = 7) and female (WT = 6; Dp16 = 6) mice. (I–J) Body temperature in the light and dark cycle of male (I) and female (J) Dp16 and WT mice on HFD. Sample size for male (WT = 15; Dp16 = 12) and female (WT = 14; Dp16 = 15) mice. All data are presented as mean ± SEM. * P<0.05; *** P<0.001; **** P<0.0001. For body weight over time, data were analyzed by 2-way ANOVA with Sidek post hoc tests.
Figure 6—figure supplement 1
ANCOVA analysis of energy expenditure in HFD-fed mice where lean mass is used as a covariate.
ANCOVA analysis of WT and Dp16 male mice across the circadian cycle (dark and light) in ad libitum fed (A), fasted (B), and refed (C) states. ANCOVA analysis of WT and Dp16 female mice across the circadian cycle (dark and light) in ad libitum fed (D), fasted (E), and refed (F) states. Male Sample size: WT = 12; Dp16 = 12. Female sample size: WT = 14; Dp16 = 14.
Figure 6—figure supplement 2
Serum Triiodothyronine (T3), sex and stress hormone levels in WT and Dp16 mice fed a high-fat diet.
(A) Serum T3 levels in male and female mice. (B) Serum testosterone levels in male mice. (C) Serum estradiol levels in female mice. (D) Serum corticosterone in male and female mice. Sample size: male WT = 9–13; male Dp16 = 11–12; female WT = 14–15; female Dp16 = 14.
Figure 6—figure supplement 3
Body and tissue weights of high-fat diet (HFD)-fed male and female mice at termination of study.
Tissues were collected from male mice (50 weeks old) after they had been fed a HFD for 34.5 weeks. Body weights and the absolute (A) and relative (B; % of body weight) weights of gWAT, iWAT, liver, and kidney in Dp16 and WT male mice. Female tissues (45 weeks old) were mice had been fed a HFD for 26 weeks. Body weights and the absolute (A) and relative (B; % of body weight) weights of gWAT, iWAT, liver, heart, and kidney in Dp16 and WT female mice. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue. Sample size: WT male = 14; Dp16 male = 12; WT female = 14; Dp16 female = 12. All data are presented as mean ± SEM. * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001.
Figure 6—figure supplement 4
Hydroxyproline (marker of fibrosis) and malondialdehyde (marker of oxidative stress) levels in the liver, gWAT, and iWAT of Dp16 mice on high-fat diet (HFD).
(A–F) Quantification of hydroxyproline content in the liver, gWAT, and iWAT of Dp16 male (A–C) and female (D–F) mice and their corresponding WT controls. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue. Sample size: male WT = 13–14 and Dp16 = 10–12; female WT = 10–14 and Dp16 = 12–14. (G–L) Quantification of malondialdehyde (MDA) levels in the liver, gWAT, and iWAT of Dp16 male (G–I) and female (J–L) mice and their corresponding WT controls. gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue. Sample size: male WT = 13–14 and Dp16 = 12; female WT = 14 and Dp16 = 13–14. All data are presented as mean ± SEM. * P<0.05; ** P<0.01.
Figure 7
Exacerbated glucose intolerance and insulin resistance in Dp16 mice fed a high-fat diet (HFD).
(A–B) Overnight fasting insulin, blood glucose, serum triglyceride, cholesterol, non-esterified free fatty acids (NEFA), and β-hydroxybutyrate (ketone) in male (A) and female (B) Dp16 and WT mice on HFD. Sample size for male mice (WT = 15; Dp16 = 12) and female mice (WT = 14; Dp16 = 14). (C–F) Exacerbated glucose intolerance as determined by the glucose tolerance test (GTT) in male (C) and female (E) Dp16 compared to WT controls on HFD. Exacerbated insulin resistance as determined by the insulin tolerance test (ITT) in male (D) and female (F) Dp16 compared to WT controls. Sample size for male mice (WT = 15; Dp16 = 12) and female mice (WT = 14; Dp16 = 14). (G–H) The rate of triglyceride clearance in response to lipid gavage as determined by the lipid tolerance test (LTT) in male (G) and female (H) Dp16 and WT mice. Sample size for male mice (WT = 15; Dp16 = 12) and female mice (WT = 14; Dp16 = 14). (I–J) Pooled mouse sera from male (I) and female (J) Dp16 and WT mice were fractionated by fast protein liquid chromatography (FPLC), and the triglyceride and cholesterol content of each fraction was quantified. Fractions corresponding to very-low density lipoprotein (VLDL), low-density lipoprotein (LDL), intermediate-density lipoprotein (IDL), and high-density lipoprotein (HDL) are indicated. All data are presented as mean ± SEM. * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001. For all tolerance tests, data were analyzed by two-way ANOVA with Sidek post hoc tests.
Tables
Table 1
Selective differential metabolites in the liver and serum of Dp16 male mice.
Metabolites are considered significantly different if fold change (FC)>1.2 or<0.833, p-value <0.05, and the variable importance in projection (VIP) score is >1. Sample size: WT (n=6) and Dp16 (n=6).
| Name | Class | log2FC | p-value | VIP | Up.Down |
|---|---|---|---|---|---|
| Liver | |||||
| Chenodeoxycholylmethionine | Bile acids | 0.906 | 0.0003 | 1.23 | Up |
| 23-Nordeoxycholic acid | Bile acids | –0.958 | 0.0066 | 1.08 | Down |
| Taurolithocholate sulfate | Bile acids | –0.796 | 0.0490 | 2.11 | Down |
| 7 a,12a-Dihydroxy-cholestene-3-one (DHCHO) | Cholestane steroids | 3.771 | 0.0123 | 2.17 | Up |
| Undecanedioylcarnitine | Acylcarnitine | 1.016 | 0.0217 | 1.68 | Up |
| (6E)-Tridec-6-enedioylcarnitine | Acylcarnitine | 0.803 | 0.0303 | 1.88 | Up |
| (2E,5Z,7E)-Decatrienoylcarnitine | Acylcarnitine | –2.315 | 0.0212 | 1.05 | Down |
| (9Z,11E,13Z)-Octadeca-9,11,13-trienoylcarnitine | Acylcarnitine | –1.277 | 0.0349 | 1.44 | Down |
| Icosadienoic acid | Fatty acids and conjugates | 2.030 | 0.0042 | 2.80 | Up |
| 12-HHTrE | Fatty acids and conjugates | –0.679 | 0.0402 | 1.56 | Down |
| LysoPC(18:4(6Z,9Z,12Z,15Z)/0:0) | Glycerophosphocholines | 0.994 | 0.0214 | 1.48 | Up |
| Lipoyllysine | Lipoamides | –1.226 | 0.0440 | 1.54 | Down |
| all-trans-4-Oxoretinoic acid | Retinoids | –1.065 | 0.0254 | 1.88 | Down |
| Sphingosine (d17:1) | Amines | 2.643 | 0.0148 | 1.70 | Up |
| Serum | |||||
| hyocholic acid | Bile acids | 2.493 | 0.0030 | 3.06 | Up |
| Taurolithocholate sulfate | Bile acids | 1.516 | 0.0109 | 3.29 | Up |
| Apocholic acid | Bile acids | 1.837 | 0.0173 | 3.29 | Up |
| Methyl cholate | Bile acids | 1.417 | 0.0182 | 3.29 | Up |
| Taurochenodeoxycholic acid (TCDCA) | Bile acids | 2.049 | 0.0195 | 2.55 | Up |
| Tauro-omega-muricholic acid | Bile acids | 1.597 | 0.0201 | 2.91 | Up |
| (3b,5b,7a,12a)–3,7,12-trihydroxy-Cholan-24-oic acid | Bile acids | 2.315 | 0.0209 | 2.50 | Up |
| Glycohyocholic acid (GHCA) | Bile acids | 1.709 | 0.0210 | 3.26 | Up |
| 3beta-Glycocholic acid | Bile acids | 1.764 | 0.0256 | 3.01 | Up |
| 7-Ketodeoxycholic acid (7-keto DCA) | Bile acids | 1.635 | 0.0340 | 2.51 | Up |
| 6,7-Diketolithocholic acid (6,7-diketo LCA) | Bile acids | 2.171 | 0.0400 | 2.18 | Up |
| 7,12-diketolithocholic acid (7,12-diketo LCA) | Bile acids | 2.068 | 0.0414 | 1.75 | Up |
| Glycoursodeoxycholic acid (GUDCA) | Bile acids | –1.308 | 0.0403 | 1.60 | Down |
| 23-Nordeoxycholic acid (23-nor- DCA) | Bile acids | –1.190 | 0.0032 | 1.18 | Down |
| 6,15-diketo-13,14-dihydro Prostaglandin F1alpha | Eicosanoids | 1.173 | 0.0008 | 2.20 | Up |
| Prostaglandin B1 | Eicosanoids | 2.094 | 0.0020 | 1.89 | Up |
| Prostaglandin D2 | Eicosanoids | 1.157 | 0.0041 | 1.28 | Up |
| 11-Dehydro-thromboxane B2 | Eicosanoids | 1.181 | 0.0116 | 1.08 | Up |
| 8-Isoprostaglandin F2a | Eicosanoids | 0.916 | 0.0237 | 1.10 | Up |
| Non-7-enoylcarnitine | Acyl-carnitine | –1.563 | 0.0007 | 1.79 | Down |
| 3,6-Dihydroxydecanoylcarnitine | Acyl-carnitine | –2.095 | 0.0008 | 1.83 | Down |
| (6E)-Tridec-6-enedioylcarnitine | Acyl-carnitine | 0.553 | 0.0221 | 1.16 | Up |
| trans-2-Dodecenoylcarnitine | Acyl-carnitine | 0.501 | 0.0276 | 1.24 | Up |
| O-dodecanedioylcarnitine | Acyl-carnitine | 0.663 | 0.0263 | 1.46 | Up |
| 7-Keto-dehydroepiandrosterone | Androstane steroids | 1.122 | 0.0005 | 1.70 | Up |
| Testosterone | Androstane steroids | 1.984 | 0.0081 | 2.99 | Up |
| Dehydroepiandrosterone (DHEA) | Androstane steroids | 2.470 | 0.0256 | 3.12 | Up |
| 5Alpha-Androstan-17-Beta-Ol-3-One (DHT) | Androstane steroids | 1.544 | 0.0368 | 3.48 | Up |
| 19-Hydroxyandrost-4-ene-3,17-dione | Androstane steroids | 1.781 | 0.0404 | 2.10 | Up |
| Androstenedione | Androstane steroids | 0.441 | 0.0486 | 1.49 | Up |
| FAHFA(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/14-O-22:6(4Z,7Z,10Z,13Z,16Z,19Z)) | Fatty acids and conjugates | –1.123 | 0.0305 | 1.99 | Down |
| PC(20:4(6Z,8E,10E,14Z)–2OH(5 S,12R)/2:0) | Phospholipid | 2.498 | 0.0005 | 1.75 | Up |
| LysoPC(22:4(7Z,10Z,13Z,16Z)/0:0) | Phospholipid | 0.766 | 0.0008 | 1.89 | Up |
| LysoPC(18:4(6Z,9Z,12Z,15Z)/0:0) | Phospholipid | 1.590 | 0.0138 | 3.04 | Up |
| PC(MonoMe (11,3)/MonoMe (1,3)) | Phospholipid | 2.072 | 0.0228 | 3.07 | Up |
| 1,2-Dilauroyl-sn-glycero-3-phosphocholine | Phospholipid | –1.042 | 0.0055 | 1.31 | Down |
| LysoPE(0:0/15:0) | Phospholipid | –1.353 | 0.0087 | 1.72 | Down |
| 1-Heptadecanoyl-glycero-3-phosphoethanolamine | Phospholipid | –0.950 | 0.0116 | 1.64 | Down |
| LysoPE(20:5(5Z,8Z,11Z,14Z,17Z)/0:0) | Phospholipid | –1.087 | 0.0478 | 1.89 | Down |
| 19-Nordeoxycorticosterone | Hydroxysteroids | 1.447 | 0.0043 | 1.01 | Up |
| Lipoamide | Lipoamides | –1.707 | 0.0162 | 1.31 | Down |
Table 2
Selective differential metabolites in the liver and serum of Dp16 female mice.
Metabolites are considered significantly different if fold change (FC)>1.2 or<0.833, p-value <0.05, and the variable importance in projection (VIP) score is >1. Sample size: WT (n=6) and Dp16 (n=6).
| Name | Class | log2FC | p-Value | VIP | Up.Down |
|---|---|---|---|---|---|
| Liver | |||||
| 23-Norcholic acid (23-NCA) | Bile acids | –4.557 | 0.000008 | 4.86 | Down |
| 6,7-Diketolithocholic acid | Bile acids | –2.049 | 0.0086 | 1.56 | Down |
| 3-Oxo-7-hydroxychol-4-enoic acid | Bile acids | –1.817 | 0.0159 | 1.21 | Down |
| Apocholic acid | Bile acids | –2.577 | 0.0186 | 1.31 | Down |
| 23-Nordeoxycholic acid (23-NDCA) | Bile acids | –2.563 | 0.0350 | 2.72 | Down |
| (3b,5b,7a,12a)–3,7,12-trihydroxy-Cholan-24-oic acid | Bile acids | –3.375 | 0.0425 | 2.05 | Down |
| 20-Hydroxy-leukotriene E4 | Eicosanoids | 1.214 | 0.0261 | 2.66 | Up |
| 6-Keto-prostaglandin E1 | Eicosanoids | 0.886 | 0.0282 | 1 | Up |
| Prostaglandin E1 | Eicosanoids | 0.822 | 0.0356 | 1.56 | Up |
| Prostaglandin A1 | Eicosanoids | 0.721 | 0.0382 | 1.16 | Up |
| 4-Hydroxydecanedioylcarnitine | Acyl-carnitine | 1.690 | 0.0003 | 1.41 | Up |
| 3-Oxobutanoylcarnitine | Acyl-carnitine | 0.788 | 0.0100 | 1.1 | Up |
| 4-Hydroxyhexanoycarnitine | Acyl-carnitine | 0.701 | 0.0182 | 2.29 | Up |
| (3E)-Glutaconylcarnitin | Acyl-carnitine | 1.654 | 0.0204 | 1.27 | Up |
| O-(17-Carboxyheptadecanoyl)carnitine | Acyl-carnitine | 1.722 | 0.0262 | 1.55 | Up |
| (6E)-Tridec-6-enedioylcarnitine | Acyl-carnitine | 0.663 | 0.0278 | 1.61 | Up |
| LysoPE(22:5(7Z,10Z,13Z,16Z,19Z)/0:0) | Phospholipid | 1.792 | 0.0151 | 2.61 | Up |
| PC(MonoMe (11,3) /MonoMe (11,3)) | Phospholipid | –5.980 | 0.0054 | 2.17 | Down |
| TG(20:3n6/O-18:0/18:3(9Z,12Z,15Z)) | Triacylglycerols | 1.293 | 0.0008 | 1.68 | Up |
| Serum | |||||
| 3-Oxo-7-hydroxychol-4-enoic acid (7-HOCA) | Bile acids | 5.645 | 1.7E-07 | 2.84 | Up |
| 3beta-Glycocholic acid | Bile acids | 3.727 | 1.8E-06 | 2.46 | Up |
| Taurochenodeoxycholic acid (TCDCA) | Bile acids | 5.069 | 4.0E-06 | 2.75 | Up |
| Tauro-omega-muricholic acid | Bile acids | 5.682 | 6.9E-06 | 2.72 | Up |
| Glycohyocholic acid (GHCA) | Bile acids | 3.525 | 1.2E-05 | 2.46 | Up |
| Taurolithocholic acid (TLCA) | Bile acids | 6.759 | 2.6E-05 | 3.26 | Up |
| lithocholic acid (LCA) | Bile acids | 3.436 | 2.8E-04 | 2.49 | Up |
| Cholan-24-oic acid, 12-hydroxy-3-(sulfooxy)-, disodium salt, (3alpha,5beta,12alpha)- (9 CI) | Bile acids | 1.948 | 9.3E-04 | 1.98 | Up |
| 3beta-Hydroxy-5-cholestenoic acid | Bile acids | 2.044 | 9.0E-03 | 1.54 | Up |
| 3alpha,7alpha-Dihydroxy-12-oxo-5beta-cholanate | Bile acids | 1.725 | 1.1E-02 | 1.46 | Up |
| Glycochenodeoxycholate-3-sulfate (GCDCA-S) | Bile acids | 1.989 | 1.2E-02 | 2.19 | Up |
| (3b,5b,7a,12a)–3,7,12-trihydroxy-Cholan-24-oic acid | Bile acids | 3.245 | 1.7E-02 | 1.47 | Up |
| Beta-Hyodeoxycholic acid (β-HDCA) | Bile acids | 3.311 | 2.9E-02 | 1.86 | Up |
| hyocholic acid (HCA) | Bile acids | 1.800 | 3.8E-02 | 1.66 | Up |
| 23-Norcholic acid (23-NCA) | Bile acids | –2.306 | 3.5E-03 | 1.82 | Down |
| 23-Nordeoxycholic acid (23-NDCA) | Bile acids | –1.923 | 4.0E-02 | 1.58 | Down |
| Biliverdin | Bilirubins | 0.885 | 1.9E-02 | 2.64 | Up |
| 5,6-Dihydroxyprostaglandin F1a | Eicosanoids | 0.924 | 2.5E-03 | 1.06 | Up |
| 2-glyceryl-11,12-EET | Eicosanoids | 1.295 | 1.1E-02 | 1.39 | Up |
| 11-Dehydro-thromboxane B2 | Eicosanoids | 1.654 | 3.9E-02 | 1.38 | Up |
| 13,14-dihydro-15-keto-PGA2 | Eicosanoids | –1.304 | 1.4E-04 | 2.18 | Down |
| 15(S)-HETrE | Eicosanoids | –1.517 | 5.0E-04 | 2.63 | Down |
| THROMBOXANE B2 | Eicosanoids | –1.040 | 3.0E-03 | 1.03 | Down |
| FAHFA(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/14-O-22:6(4Z,7Z,10Z,13Z,16Z,19Z)) | Fatty acids and conjugates | –2.148 | 2.7E-05 | 2.92 | Down |
| FAHFA 38:5 | Fatty acids and conjugates | –2.415 | 3.1E-05 | 1.62 | Down |
| Arachidonic acid | Fatty acids and conjugates | –0.928 | 2.8E-04 | 2 | Down |
| N-Palmitoyl Glutamine | Fatty acids and conjugates | –1.304 | 5.2E-03 | 1.33 | Down |
| Resolvin D1 | Fatty acids and conjugates | –0.876 | 9.8E-03 | 1.44 | Down |
| LysoPC(22:4(7Z,10Z,13Z,16Z)/0:0) | Phospholipid | 0.951 | 1.9E-04 | 1.96 | Up |
| LysoPC(22:5(7Z,10Z,13Z,16Z,19Z)/0:0) | Phospholipid | 0.762 | 1.0E-03 | 1.56 | Up |
| LysoPC(22:5(4Z,7Z,10Z,13Z,16Z)/0:0) | Phospholipid | 0.654 | 2.9E-03 | 1.28 | Up |
| PC(MonoMe (11,3)/MonoMe (11,3)) | Phospholipid | 6.452 | 8.6E-03 | 1.9 | Up |
| 1-O-Palmitoyl-2-O-acetyl-sn-glycero-3-phosphorylcholine | Phospholipid | 0.816 | 3.2E-02 | 1.2 | Up |
| LysoPC(18:4(6Z,9Z,12Z,15Z)/0:0) | Phospholipid | 1.148 | 3.7E-02 | 1.47 | Up |
| LysoPA(20:3(8Z,11Z,14Z)/0:0) | Phospholipid | –0.934 | 4.3E-03 | 2.15 | Down |
| LysoPC(0:0/18:1(9Z)) | Phospholipid | –1.494 | 1.5E-03 | 1.14 | Down |
| PC(20:3(8Z,11Z,14Z)/18:3(9Z,12Z,15Z)) | Phospholipid | –0.868 | 7.3E-03 | 1.54 | Down |
| LysoPE(P-18:1(9Z)/0:0) | Phospholipid | –1.305 | 9.8E-04 | 1.18 | Down |
| Glycerophospho-N-Oleoyl Ethanolamine | Phospholipid | –1.189 | 1.7E-03 | 1.32 | Down |
| 1-heptadecanoyl-glycero-3-phosphoethanolamine | Phospholipid | –1.071 | 2.4E-03 | 1.57 | Down |
| LPE(14:0) | Phospholipid | –1.001 | 7.4E-03 | 1.01 | Down |
| LysoPE(20:3(8Z,11Z,14Z)/0:0) | Phospholipid | –0.987 | 9.4E-03 | 1.07 | Down |
| LysoPE(20:5(5Z,8Z,11Z,14Z,17Z)/0:0) | Phospholipid | –1.067 | 9.9E-03 | 1.55 | Down |
| Pregnenolone | Pregnane steroids | –1.031 | 5.7E-04 | 1.18 | Down |
| 17alpha-Hydroxyprogesterone | Pregnane steroids | –0.807 | 3.9E-03 | 2 | Down |
| Dehydroepiandrosterone sulfate (DHEAS) | Sulfated steroids | –1.334 | 4.6E-04 | 1.31 | Down |
Additional files
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Gene dosage imbalance disrupts systemic metabolism in the Dp16 Down syndrome mouse model
eLife 15:RP110476.
https://doi.org/10.7554/eLife.110476.3
