Journal: Science Advances

Article Title: Nanomedicine targeting PPAR in adipose tissue macrophages improves lipid metabolism and obesity-induced metabolic dysfunction

doi: 10.1126/sciadv.ads3731

Figure Lengend Snippet: ( A ) Dextran distribution by PET (color) overlay with x-ray computed tomography (CT; black and white). A male DIO mouse was imaged 1, 4, or 24 hours after left-side intraperitoneal injection of 30-nm dextran-NOTA- 64 Cu radiochelate. PET/CT coronal planes point show distribution in left perirenal (PR) and left gonadal (GD) AT. 3D, three-dimensional. ( B ) Tissue distribution of four sizes of dextran-NOTA- 64 Cu 24 hours after left-side intraperitoneal injection in male DIO mice by gamma well counting. Tissues include heart, blood, liver, subcutaneous AT (SubQ), and visceral AT depots: left and right PR, left and right GD, and mesenteric (Mes) ( n = 4 to 6). Data for 3.5-nm dextran were previously published . ( C ) D-PPAR [30-nm tetramethyl rhodamine isothiocyanate (TRITC) conjugate] is specific to CD11b + immune cells from AT in DIO mice by flow cytometry. ( D ) D-PPAR drug release mechanism. ( E ) Hydrodynamic diameter of dextran and D-PPAR by dynamic light scattering. ( F ) Drug release from D-PPAR in phosphate-buffered saline (PBS) at 37°C. ( G ) Metabolic gene expression in RAW264.7 MΦs polarized to the M1 phenotype by lipopolysaccharide (LPS) and interferon-γ (IFN-γ) shows up-regulation of Sirt1 , Cd36 , Ucp2 , Ucp3 , and Ifn β after treatment with D-PPAR or F-PPAR after 24 hours. Expression is measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) normalized to the housekeeping gene Tbp by the ΔΔCt method. Letters indicate P < 0.05 for mean differences by one-way analysis of variance (ANOVA) with Tukey’s post hoc test. Asterisks are included as a redundant notation of letter conventions that indicate groups with significant differences, i.e., “a” indicates significant difference from “b” but neither “a” nor “b” is significantly different from “ab” . Data are presented as mean ± SEM ( n = 3). Rel., relative.

Article Snippet: For analysis of dextran localization , tissues were then blocked with 5% donkey serum (Jackson ImmunoResearch, West Grove, PA) and 1% Triton X-100 in PBS before labeling with a rat anti-mouse antibody against CD11b (1:200; Thermo Fisher Scientific; table S2) for 1 hour at room temperature.

Techniques: Computed Tomography, Injection, Positron Emission Tomography-Computed Tomography, Flow Cytometry, Saline, Gene Expression, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Science Advances

Article Title: Nanomedicine targeting PPAR in adipose tissue macrophages improves lipid metabolism and obesity-induced metabolic dysfunction

doi: 10.1126/sciadv.ads3731

Figure Lengend Snippet: MΦ populations are reduced in visceral AT after 1-week treatment with D-PPAR, including ( A ) CD45 + CD11b + , ( B ) CD45 + CD11b + Ly6C + , ( C ) CD45 + CD11b + Ly6C − F4/80 + , and ( D ) CD45 + CD11b + Ly6C − F4/80 + CD9 + by flow cytometry. ( E ) Flow cytometry gating strategy for leukocytes (CD45), myeloid cells (CD11b), and myeloid subtype markers Ly6C, F4/80, and CD9. Full gating strategy is in fig. S8A and described in Materials and Methods. FSC-A, forward scattering area. PE, phycoerythrin. ( F to I ) Neutral lipid content in MΦ subtypes following F-PPAR or D-PPAR treatment measured by mean fluorescence intensity (MFI) of BODIPY by flow cytometry. Values correspond to each cell population in panels [(A) to (D)]. Example dataset is in fig. S8A and described in Materials and Methods. ( J ) Neutral lipid content in SVF cells treated with D-PPAR in vitro, showing confocal fluorescence micrograph of BODIPY dye (green) with nuclear stain (blue). Quantification is in fig. S6. ( K ) Confocal fluorescence micrograph of whole-mount AT of mice treated with TRITC–D-PPAR, showing colocalization of AT MΦs (red), TRITC (white), and small lipid droplets (BODIPY; green). Representative image from untreated control is shown in fig. S9. ( L ) TRITC–D-PPAR uptake corresponds to lipid-rich cells (BODIPY) in SVF CD45 + CD11b + populations measured by flow cytometry. Results are from the same experiment as in (K). Example dataset is in fig. S8B and described in Materials and Methods. ( M ) One-week D-PPAR treatment reduces EV secretion by AT resected from DIO mice via nanoparticle tracking analysis (NTA; scattering). ( N ) EVs did not differ in size. ( O and P ) F-PPAR and D-PPAR reduced lipid-laden EVs (NTA; BODIPY fluorescence). Different letters denote significance of P < 0.05 by one-way ANOVA with Tukey’s post hoc test ( n = 3 to 4). See legend for additional description of statistical notation.

Article Snippet: For analysis of dextran localization , tissues were then blocked with 5% donkey serum (Jackson ImmunoResearch, West Grove, PA) and 1% Triton X-100 in PBS before labeling with a rat anti-mouse antibody against CD11b (1:200; Thermo Fisher Scientific; table S2) for 1 hour at room temperature.

Techniques: Flow Cytometry, Fluorescence, In Vitro, Staining, Control