Adipocyte release of nucleosides contributes to obesity-associated inflammation
Data files
Dec 03, 2025 version files 60.77 MB
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Fig_1A_3T3.csv
4.12 KB
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Fig_1B_Primary_Mouse_A_SVF.csv
2.63 KB
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Fig_1D_hypertrophic.csv
1.83 KB
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Fig_2A_Mouse_Chips.csv
4.45 KB
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Fig_2B_Mouse_adipose_IF.csv
1.48 KB
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Fig_3A_primary_human_adipocytes_fibroblasts.csv
4.95 KB
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Fig_3B_Human_adipose_chip.csv
5.47 KB
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Fig_4A_adipose_ENT_expression.csv
1.11 KB
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Fig_4B_Adipocyte_vs_SVF_ENT1.csv
220 B
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Fig_5A_MS5_3T3_ENTi.csv
7.33 KB
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Fig_5B_mouse_adipose_IF_ENTi.csv
1.36 KB
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Fig_6A_mouse_adipose_cytokine_expression.csv
1.35 KB
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Fig_6B_mouse_macrophage_cytokine_expression.csv
1.70 KB
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Fig_S1_mouse_SVF_adipocyte_nucleosides.csv
4.61 KB
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Fig_S2_mouse_plasma_nucleosides.csv
889 B
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Figure_S1.tif
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README.md
9.77 KB
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Subject1_20x_1_CAV1.tif
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Subject1_20x_1_DAPI.tif
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Subject1_20x_1_ENT1.tif
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Subject1_20x_1_Lipidtox.tif
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Subject1_20x_1_Merge.tif
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Subject2_10x_CAV1.tif
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Subject2_10x_DAPI.tif
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Subject2_10x_ENT1.tif
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Subject2_10x_Merge.tif
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Subject2_20x_CAV1.tif
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Subject2_20x_DAPI.tif
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Subject2_20x_ENT1.tif
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Subject2_20x_Lipidtox.tif
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Subject2_20x_Merge.tif
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Subject3_20x_CAV1.tif
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Subject3_20x_DAPI.tif
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Subject3_20x_ENT1.tif
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Subject3_20x_Lipidtox.tif
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Subject3_20x_Merge.tif
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Abstract
Obese adipose tissue drives inflammation through several mechanisms. Extracellular nucleosides can induce macrophage inflammation through the activation of toll-like and purinergic receptors. The present study quantified nucleoside release from adipocytes and adipose tissue. Cultured mouse adipocytes released many nucleosides used in RNA/DNA. Adipose tissue from obese mice released more nucleosides than that from control non-obese mice, resulting in higher interstitial fluid concentrations. Consistent with the mouse study, human adipose tissue also showed significant release of adenosine/deoxyadenosine, guanosine/deoxyguanosine, and uridine ex vivo. Adipocytes release nucleosides in part through the equilibrative nucleoside transporter 1 (ENT1), and this release induces macrophage TNFα and IL-1β expression. Treatment of mice with the ENT1 inhibitor in vivo increased adipose tissue interstitial nucleoside concentrations and cytokine expression. These data uncover a previously unknown phenomenon of adipocyte release of nucleosides, which contribute to adipose tissue inflammation in obesity.
Dataset DOI: 10.5061/dryad.bk3j9kdp3
Description of the data and file structure
Cells were cultured in various conditions, and nucleoside release was quantified by LC-MS/MS-MRM. Mouse and human adipose tissue explants were cultured in a PDSM flow chamber, and nucleoside release was quantified in the effluent. Expression of nucleoside transporter ENT1 was quantified by qPCR and Western blot in human adipose tissue. Mice were treated with the ENT1 inhibitor, and adipose tissue interstitial fluid collected for measure of nucleoside concentration. Adipose tissue was also collected for cytokine expression. Murine macrophages were treated with adipocyte conditioned media or guanosine and cytokine expression quantified with qPCR
Files and variables
File: Fig_1A_3T3.csv
Description: 3T3-L1 fibroblasts and adipocytes were cultured and media release of nucleosides quantified over 48 hours
Variables
- Cell counts: Number of cells in control well for normalization
- Condition (48hrs): media, adipocytes, or fibroblasts
- Measure: concentration, net release (subtracting media alone), net release per 10^5 cells
- Units: micromolar, micromolar/10^5 cells
File: Fig_1B_Primary_Mouse_A_SVF.csv
Description: Mouse adipose tissue derived fibroblasts and adipocytes were cultured and media release of nucleosides quantified over 48 hours
Variables
- Cell counts: Number of cells in control well for normalization
- Condition (48hrs): media, adipocytes, or fibroblasts
- Measure: concentration, net release (subtracting media alone), net release per 10^5 cells
- micromolar, micromolar/10^5 cells
File: Fig_1D_hypertrophic.csv
Description: Mouse adipose tissue derived mature and hypertrophic adipocytes were cultured and media release of nucleosides quantified over 48 hours
Variables
- Cell type: mature vs. hypertrophic adipocytes
- technical replicates: replicates or fold change
- Measure: concentration, net release (subtracting media alone), net release per 10^5 cells
- micromolar, fold change over mature
File: Fig_2A_Mouse_Chips.csv
Description: Adipose tissue from mice was removed and cultured in PDSM flow chambers. Released nucleosides were measured over time in effluent media
Variables
- Diet: control vs. obese
- Run time (hour): hours of media collection
- Measure: micromolar, area under the curve
File: Fig_2B_Mouse_adipose_IF.csv
Description:
Variables
- Group: control vs. obese mice
- Excluded: x indicates which samples were excluded. Reason for exclusion noted below
- Measure: concentration in uM
File: Fig_3A_primary_human_adipocytes_fibroblasts.csv
Description: Human adipose tissue SVF adipocytes and fibroblasts were cultured and media release of nucleosides quantified over 48 hours
Variables
- Cell counts: Number of cells in control well for normalization
- Condition (48hrs): media, adipocytes, or fibroblasts
- Measure: concentration, net release (subtracting media alone), net release per 10^5 cells
- micromolar, micromolar/10^5 cells
File: Fig_3B_Human_adipose_chip.csv
Description: Human adipose tissue explants were minced and put into PDMS flow chamber. Nucleoside release was quantified in media effluent over time
Variables
- Sample
- Run time: in hours
- Nucleoside concentrations: in micromolar
File: Fig_4A_adipose_ENT_expression.csv
Description: Human adipose tissue explants were lysed for RNA
Variables
- TPCL#: subject ID
- BMI: body mass index in kg/m^2
- Group: obese vs. nonobese
- Site: anatomical site of adipose tissue
- Measure: RQ
- ENT1-4: RQ of expression
File: Fig_4B_Adipocyte_vs_SVF_ENT1.csv
Description: Human adipose tissue explants were digested and adipocytes separated from SVF
Variables
- Subject: subject ID
- BMI: body mass index in kg/m^2
- Measure: ENT1 RQ
- Adipocytes: ENT1 expression in adipocytes
- SVF: ENT1 expression in SVF
File: Fig_5A_MS5_3T3_ENTi.csv
Description: Adipocytes and MS5 cells were cultured alone or with ENT inhibitor. Nucleoside concentrations were measured over 72 hours
Variables
- Cell: 3T3-L1 adipocytes or MS5 cells
- Condition: treatment with control media, ENTi, or NBMPR
- Time: 0, 24, 48, or 72 hours
- Nucleosides: nucleoside concentration in micromolar
File: Fig_5B_mouse_adipose_IF_ENTi.csv
Description: Obese mice were treated with ENTi or vehicle. Adipose interstitial fluid was collected and nucleosides measured
Variables
- Drug: ENTCi vs. vehicle
- Excluded: x indicates samples excluded for undetectible levels
- Nucleoside concentrations in micromolar
File: Fig_6A_mouse_adipose_cytokine_expression.csv
Description:
Variables
- Diet: Nonobese vs. Obese
- Treatment: Vehicle vs. ENTi
- Cytokine dCT: dCT of respective cytokine
- Cytokine dCT ave: dCT of control condition of respective cytokine
- Cytokine RQ: RQ of respective cytokine
File: Fig_6B_mouse_macrophage_cytokine_expression.csv
Description: Mouse macrophages were treated with control media, guanidine, or adipocyte conditioned media
Variables
- cytokine: IL6, TNFa, or IL1b
- NT DCt: no treatment DCt
- Treatment: control, ACM, or rG
- ssRNA: present (+) or absent (-)
File: Fig_S1_mouse_SVF_adipocyte_nucleosides.csv
Description: Mouse adipose tissue SVF cells were derived from control and obese mice and differentiated into adipocytes. Nucleoside release was then quantified in the media
Variables
- Diet: Nonobese vs. obese
- Measure: Concentration (micromolar), Net release over media alone (micromolar), or net release per 105 cells (micromolar/105 cells)
- Excluded: x indicates sample excluded due to outliers
- Cell counts (X10^5): cell counts
- Nucleoside: nucleoside concentrations in units indicated
File: Fig_S2_mouse_plasma_nucleosides.csv
Description: Plasma from control and obese mice was assayed for nucleoside concentration
Variables
- Diet: control vs. obese mice
- nucleoside: Nucleoside concentration in micromolar
Fike: Figure_S1.tif
A: Nucleoside release from cultured adipocytes derived from control (n=3) and obese (HFD; n=5) mice. B: Nucleoside levels in plasma from control and obese mice (n=6).
File: Subject1_20x_1_CAV1.tif
Description: IHC image of adipose tissue at 20X magnification from subject 1 stained for caveolin 1
File: Subject1_20x_1_DAPI.tif
Description: IHC image of adipose tissue at 20X magnification from subject 1 stained for DAPI
File: Subject1_20x_1_Lipidtox.tif
Description: IHC image of adipose tissue at 20X magnification from subject 1 stained for lipid
File: Subject1_20x_1_ENT1.tif
Description: IHC image of adipose tissue at 20X magnification from subject 1 stained for ENT1
File: Subject1_20x_1_Merge.tif
Description: IHC image of adipose tissue at 20X magnification from subject 1 all channels merged
File: Subject2_20x_CAV1.tif
Description: IHC image of adipose tissue at 20X magnification from subject 2 stained for caveolin 1
File: Subject2_20x_DAPI.tif
Description: IHC image of adipose tissue at 20X magnification from subject 2 stained for DAPI
File: Subject2_20x_Lipidtox.tif
Description: IHC image of adipose tissue at 20X magnification from subject 2 stained for lipid
File: Subject2_20x_ENT1.tif
Description: IHC image of adipose tissue at 20X magnification from subject 2 stained for ENT1
File: Subject2_20x_Merge.tif
Description: IHC image of adipose tissue at 20X magnification from subject 2 all channels merged
File: Subject2_10x_CAV1.tif
Description: IHC image of adipose tissue at 10X magnification from subject 2 stained for caveolin 1
File: Subject2_10x_DAPI.tif
Description: IHC image of adipose tissue at 10X magnification from subject 2 stained for DAPI
File: Subject2_10x_ENT1.tif
Description: IHC image of adipose tissue at 10X magnification from subject 2 stained for ENT1
File: Subject2_10x_Merge.tif
Description: IHC image of adipose tissue at 10X magnification from subject 2 all channels merged
File: Subject3_20x_CAV1.tif
Description: IHC image of adipose tissue at 20X magnification from subject 3 stained for caveolin 1
File: Subject3_20x_DAPI.tif
Description: IHC image of adipose tissue at 20X magnification from subject 3 stained for DAPI
File: Subject3_20x_Lipidtox.tif
Description: IHC image of adipose tissue at 20X magnification from subject 3 stained for lipid
File: Subject3_20x_ENT1.tif
Description: IHC image of adipose tissue at 20X magnification from subject 3 stained for ENT1
File: Subject3_20x_Merge.tif
Description: IHC image of adipose tissue at 20X magnification from subject 3 all channels merged
File: Supplementary_Tables.doc
Description: Supplementary tables from manuscript. Note: in table S1, biospecimen number is a number assigned to each specimen that cannot be tracked back to individual patients, and therefore is not considered a direct identifier. Specimens were released to laboratories in a de-identified manner.
Variables
- Diet: control vs. obese mice
- nucleoside: Nucleoside concentration in micromolar
Human subjects data
Subjects undergoing procedures at UCLA signed consent to allow any remnant specimens to be de-identified and used in research. Identifying data was not attached to these remnants, which were distributed in a de-identified manner.