Impaired adipogenesis and lipid storage capacity in subcutaneous adipose tissue of patients with PMOS
Adeline Divoux, Edina Erdos, Katie L. Whytock, Timothy F. Osborne, Steven R. Smith
Adeline Divoux, Edina Erdos, Katie L. Whytock, Timothy F. Osborne, Steven R. Smith
View: Text | PDF
Research Article Cell biology Metabolism

Impaired adipogenesis and lipid storage capacity in subcutaneous adipose tissue of patients with PMOS

Abstract

Women with PMOS (formally termed PCOS) have an overall increased prevalence of metabolic syndrome (MetS) and central obesity. To help determine whether there might be changes in s.c. adipose tissue (SAT) associated with these abnormalities, we performed single-nuclei and scRNA-seq on SAT biopsies from 15 premenopausal PMOS women with signs of insulin resistance and 17 healthy BMI-matched controls. In SAT from PMOS versus control we observed a higher ratio of fibrotic versus insulin sensitive adipocytes and a higher ratio of mesenchymal stem cells (MSCs) to preadipocytes. Further in silico analysis suggested that preadipocytes in PMOS are more inflammatory and have a reduced capacity for differentiation. Slit homolog 2 (SLIT2), which is expressed at higher levels in MSC from PMOS, decreased adipogenesis in cell culture assays likely through its interaction with the Roundabout homolog 1 and homolog 2 (ROBO1/2) receptor expressed on the surface of preadipocytes. These new observations are consistent with higher SLIT/ROBO signaling, leading to reduced differentiation in the SAT of PMOS as an underlying mechanism for the aberrant ectopic fat accumulation and the development of MetS in PMOS.

Authors

Adeline Divoux, Edina Erdos, Katie L. Whytock, Timothy F. Osborne, Steven R. Smith

×

Figure 6

Potential interaction between MSC and Preadipocytes via SLIT2/ROBO pathway.

Options: View larger image (or click on image) Download as PowerPoint
Potential interaction between MSC and Preadipocytes via SLIT2/ROBO pathw...
(A) Level of differentiation of ABD-ADSCs after 12 days of incubation of adipogenic differentiation cocktail supplemented with testosterone (T). Oil red O was used to quantify the lipid accumulation at day 12. n = 10 ADSCs, isolated from 5 CTRL and 5 PMOS. Representative pictures of CTRL ADSCs at D12 of differentiation with or without testosterone in the media. Scale bars: 200 μm. (B) Circle plots of secreted signaling pathways coming from MSC population enriched in CTRL and in PMOS group. The colors of the circle specify which cell type receives the signal. (C) Circle plots of ECM-receptor signaling pathways enriched in MSC CTRL and in MSC PMOS group. The colors of the circle specify which cell type receives the signal. (D) Violin plots comparing the expression level of SLIT2, ROBO1, and ROBO2 in MSC, Pread-1, and Pread-2 nuclei between CTRL and PMOS group. (E) Immunofluorescence staining of s.c. adipose tissue using WGA (488 excitation length of the fluorochrome) and ROBO1 (647 excitation length of the fluorochrome) antibodies. Hoescht was used to stain the nuclei. Scale bar: 200 μm. (F) Immunofluorescence staining of stroma vascular fraction cells using ZNF423 (488 excitation length of the fluorochrome) and ROBO1 (647 excitation length of the fluorochrome) antibodies. Hoescht was used to stain the nuclei. Original magnification, ×20. (G) Violin plots comparing the expression level of SLIT2, ROBO1, and ROBO2 in scRNA-seq dataset. (H) Histogram of SLIT2 secretion in 24-hour-condition media of MSC isolated from CTRL (n = 5) and from PMOS (n = 5). **P < 0.05 by Mann-Whitney U test. (I) Expression of PPARG, CEBPA, and PLIN1 in cells treated with SLIT2 one day before the beginning of differentiation and during 4 days of differentiation. Cells were collected at day 4, RNA was extracted, and gene expressions determined by qPCR. See also Supplemental Figure 5. **P < 0.01 by Wilcoxon matched-pairs signed rank test.