White and beige adipocytes in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) are maintained by proliferation and differentiation of adipose progenitor cells (APCs). Here we use mice with tissue-specific telomerase reverse transcriptase (TERT) gene knockout (KO), which undergo premature telomere shortening and proliferative senescence in APCs, to investigate the effect of over-nutrition on APC exhaustion and metabolic dysfunction. We find that TERT KO in the Pdgfra+ cell lineage results in adipocyte hypertrophy, inflammation and fibrosis in SAT, while TERT KO in the Pdgfrb+ lineage leads to adipocyte hypertrophy in both SAT and VAT. Systemic insulin resistance is observed in both KO models and is aggravated by a high-fat diet. Analysis of human biopsies demonstrates that telomere shortening in SAT is associated with metabolic disease progression after bariatric surgery. Our data indicate that over-nutrition can promote APC senescence and provide a mechanistic link between ageing, obesity and diabetes.
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We thank S. H. Pan for the TERT floxed mice and V. Lindner for Pdgfrb-Cre mice. We thank Z. Mao for help with microscopy. We also thank A. Ribas-Latre and E. Sahin for expert advice. This work was supported by the Harry E. Bovay, Jr. Foundation. Z.Z. and Y.D. were partially supported by the Cancer Genomics Core funded by the Cancer Prevention and Research Institute of Texas (CPRIT) (RP180734) and National Institutes of Health grant (R01LM012806).
The authors declare no competing interests.
Peer review information Primary handling editor: Christoph Schmitt. Nature Metabolism thanks Thomas von Zglinicki and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
a, TRF assay reveals advanced telomere shortening in SAT of both a-KO males (M) and females (F) at 8 months of age. b, Telo-FISH staining (red) on sections from Pdgfra-Cre;mTmG;TERTfl/fl (WT) and Pdgfra-Cre;mTmG;TERTfl/fl male mice reveals normal telomeres in mG+ cells of brain and muscle. c, Telo-FISH staining on culture-plated cells from VAT and SAT of control Pdgfra-Cre; mTmG and Pdgfra-Cre; mTmG; TERTfl/fl mice reveals advanced telomere shortening (less red signal) in SAT mG+ cells of a-KO mice. d, Telo-FISH staining on SAT sections from mice in c reveals telomere shortening and nuclear p16 IF signal in mG+ cells of a-KO mice. e, q-PCR reveals telomere shortening in SAT of 8-month old WT mice compared to 3-month old WT mice. Evident is more advanced telomere shortening in SAT of 8-month old α-KO males (M) and females (F), compared to 8-month old WT males and females, respectively. f, Quantitative RT-PCR analysis reveals higher p16 and p21 mRNA expression in VAT and SAT of a-KO mice, compared to WT mice, at 6 months of age. Data are normalized to 18S RNA. N = 3 mice / group. Scale bar=50 µm in all panels. P: Student’s t-test (2-sided).
a, Quantitative RT-PCR demonstrates higher Pdgfa mRNA expression in VAT and SAT of 6 month old KO males, compared to 6 month old WT males. Data are normalized to 18S RNA. b, Quantitative RT-PCR demonstrates higher Pdgfa mRNA expression in SVF from SAT of 3 month-old KO males, which is not accompanied by increased p16 and p21 expression at that age. In contrast, in SVF from SAT of 7 month-old KO males, higher Pdgfa expression is accompanied by increased p16 and p21 mRNA expression. Data are normalized to 18S RNA. N = 3 mice / group. P: Student’s t-test (2-sided).
a, qPCR analysis of DNA from whole tissue reveals telomere shortening in SAT but not in VAT of a-KO mice at 8 months of age, compared to WT mice at 8 months of age. Real-time PCR data are normalized to data for a single copy gene. b, Quantitative RT-PCR demonstrates higher p16 and TNFa mRNA expression, normalized to 18S RNA, in cultured MEFs of a-KO mice, compared to WT mice. c, EdU incorporation measurement by flow cytometry in control mTmG;TERTfl/fl and a-KO Pdgfra-Cre;mTmG; TERTfl/fl males (2-month-old, HFD-fed) and females (20-month-old, chow-fed). 4 h after EdU injection, mG+ and mT+ SVF cells were gated and EdU incorporation was compared for SAT. EdU fluorescence: 647 nm channel, side scatter (SSC) is used for separation. d, IF analysis of SAT from 12-month-old mice reveals increased infiltration of CD68+ CD206- (M1) macrophages and increased phosphorylation of NFkB subunit p65 in PDGFRα+ cells of a-KO mice, compared to WT mice. Adipocytes are positive for perilipin1 (PLN1). Nuclei are blue. e, Masson’s trichrome staining reveals increased fibrosis in livers of 12-month-old a-KO males compared to WT males. In all panels, N = 3 mice / group; P: Student’s t-test (2-sided). Scale bar=50 µm in all panels.
a, Echo MRI data demonstrating comparable fat mass in a-KO and WT males raised on HFD for indicated number of months. N = 5-6 mice / group. b, Comparable food intake by a-KO and WT males raised on HFD. N = 5-6 mice / group. c, Comparable spontaneous locomotor activity of a-KO and WT males raised on HFD. N = 5-6 mice / group. d, Comparable physical endurance, reflected by Joules of work performed, by a-KO and WT males. N = 5-6 mice / group. e, Indirect calorimetry based on oxygen consumption (VO2) and carbon dioxide production (VCO2) indicates decreased energy expenditure in a-KO males pre-fed HFD for 8 months. N = 5-6 mice / group. f, Masson’s trichrome staining of VAT from a-KO and WT males raised on HFD for 8 months. g, Telo-FISH staining (red) on SAT and VAT sections from Pdgfra-Cre;mTmG mice raised on chow or HFD revealing a higher degree of telomere shortening (less red signal) induced by DIO in mG+ cells. a-g, N = 5-6 mice / group. Scale bar=50 µm in all panels. P: Student’s t-test (2-sided).
a, Echo MRI data demonstrating lower fat mass in a-KO females raised on HFD for indicated number of months. N = 5-6 mice / group. b, H&E staining demonstrates smaller BAT adipocytes and normal liver and muscle anatomy in a-KO females raised on HFD. Scale bar=50 µm. c, Quantitative RT-PCR demonstrates a lack of p16 induction, despite higher Pdgfa mRNA expression, in SAT of 4-month old a-KO females, compared to 4 month old WT females. Data normalized to 18S RNA. N = 5-6 mice / group. P: Student’s t-test (2-sided).
a, Quantitative RT-PCR demonstrates higher senescence marker and pdgfa induction in VAT than in SAT of 8-month old b-KO males compared to WT males. Data normalized to 18S RNA. N = 5 mice / group. b, Comparable food intake by b-KO and WT males. c, Comparable spontaneous locomotor activity of b-KO and WT males. d, RER in males calculated based on oxygen consumption (VO2) and carbon dioxide production (VCO2). e, Indirect calorimetry-based measurement of energy expenditure in b-KO and WT males at 8 months of age. f, Comparable cold tolerance in b-KO and WT males placed at 4 °C. g, H&E staining does not reveal abnormality in BAT and liver of b-KO males at 8 months of age. Scale bar=50 µm. h, Echo MRI data: no body composition abnormality in b-KO females at 8 months of age. i, ITT: no insulin resistance in 8-month-old b-KO females. j, Comparable cold tolerance in b-KO and WT females placed at 4 °C. N = 6 mice / group. P: Student’s t-test (2-sided).
a, ScRNAseq data violin plots showing that Pdgfra is expressed in both Dpp4+ and Cd142+ cells, while Pdgfrb is mainly expressed in Cd142+ cells. b, UMAPs of WT, a-KO and b-KO SAT CD45- SVF cells from Fig. 7c were overlapped to identify cells expressing genes assigned in Fig. 7d. N = 4 mice / group (combined). c, Expression of Cdkn1a and Pdgfa determined by scRNAseq for combined Dpp4+ and Cd142+ ASC compared for WT, a-KO, and b-KO mice.
a, IF analysis of SAT reveals increased infiltration of PDGFRα+ (orange, anti-rat Cy5 secondary antibody) and PDGFRb+ (red, anti-rabbit Cy3 secondary antibody) cells in a-KO and b-KO 1-year-old males raised on chow compared to WT male littermates. Isolectin B4 (green) stains endothelium. Nuclei are blue. Scale bar=50 µm. Graph: the ratio of positive cells / total nucleated cells in 4-5 view fields / sample. Plotted are mean +/- SEM. P: Student’s t-test (2-sided). Analysis of tissues from 3 mice / genotype showed similar results. b, Changes in AT stroma resulting from senescence of Pdgfra and Pdgfrb lineages in VAT. UMAP clusters of cells identified by scRNAseq in VAT of TERT a-KO and b-KO mice. Shown are ASC, EC and PA only, CD45+ leukocytes are not shown. Note selective accumulation of Cd142+ ASC in VAT of b-KO mice. Note high expression of Cdkn1a and IL-6 and low expression on Ki67 in ASC.
Extended Data Fig. 9 Consequences of pharmacological adipocyte progenitor depletion in mice treated with D-WAT.
WT males pre-treated with D-WAT or PBS (control) at 4 months of age were subsequently maintained on HFD and analyzed 9 months later. a, Echo MRI data reveals higher adiposity in old mice pre-treated with D-WAT. b, H&E staining reveals larger adipocytes and fibrosis (arrow) in SAT of old mice pre-treated with D-WAT. Scale bar=50 µm. c, GTT reveals lower glucose tolerance of old mice pre-treated with D-WAT. N = 4 mice / group. *P < 0.05 (Student’s t-test, 2-sided).
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Gao, Z., Daquinag, A.C., Fussell, C. et al. Age-associated telomere attrition in adipocyte progenitors predisposes to metabolic disease. Nat Metab 2, 1482–1497 (2020). https://doi.org/10.1038/s42255-020-00320-4