Imaging of adipose tissue vasculature in GFP transgenic mice.

<p><i>A</i>: Incorporation of BMD-EC in mammary fat pad vasculature in WT/Tie2-GFP-BMT mice. <i>B</i>: <i>Ex vivo</i> confocal microscopy imaging of mammary fat pad in two mice with age-related obesity (∼18 months old). BMD-EC contributed to approximately 8% of the mammary fat pad vasculature. <i>C</i>: Representative image of GFP+ BMD-EC in the mammary fat pad of an obese mouse after 6 weeks on a high fat diet. BMD-EC contribution to mammary fat pad vasculature was minimal (0.8%, n = 4). Vessels were perfused with biotinylated-lectin and stained with streptavidin Texas Red (shown in red), while the nuclei were stained with DAPI (in blue, <i>A</i>, <i>B</i>). Rhodamine-dextran MW 2,000,000 was infused for vessel enhancement in <i>C</i>. <i>D:</i> Representative image of occasional GFP+ BMD-EC (arrow) in perfused mammary fat pad blood vessels in a 12 months old WT/Actb-GFP-BMT mouse. Vessels were perfused with biotinylated-lectin and stained with streptavidin Texas Red (red), while the nuclei were stained with DAPI (blue). Images are 1.72 mm across in <i>A</i>, 310 µm across in <i>B</i>, 700 µm across in <i>C</i>, 1.72 mm across in <i>D</i>.</p

Effects of VEGFR1 and VEGFR2 blockade in mice with DIO.

<p><i>A:</i> Body weight gain relative to weight at day 0 for mice given different diets and treatments. Male C57BL6 mice, 10–12 weeks old at time 0, were used for all groups. All diets and treatments began at day 0, at dosages and schedules as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004974#s2" target="_blank">Methods</a> section. DC101+HFD: high fat diet, DC101 treatment, white triangles. MF1+HFD: high fat diet, MF1 treatment, black squares. HFD: high fat diet controls, no treatment (n = 4) or PBS treatment (n = 4), white circles. LFD: standard diet controls, crosses. <i>B:</i> Average cross-sectional area (µm²) of adipocytes in the perigonadal fat pad at various times after beginning of high fat diet. 3–4 mice at each time point, >300 adipocytes measured for each mouse. Adipocytes in the inguinal fat pad showed a similar trend (not shown). <i>C:</i> Food intake (g/day/kg body weight) in the DC101+HFD versus HFD groups, when the DC101+HFD group was gaining less weight (days 43–91). D: Reversibility of the effects of DC101. DC101 treatment was discontinued from day 91 onward. About two weeks after cessation of treatment, the rate of weight gain in the previously treated animals resumed at a higher rate, and their body weights eventually caught up with untreated controls. All data reported as mean±sem. Asterisks denote significant difference between DC101+HFD and HFD groups (P<0.05).</p

Presentation_1_Exercise intensity governs tumor control in mice with breast cancer.pdf

IntroductionExercise is recommended as an adjunct therapy in cancer, but its effectiveness varies. Our hypothesis is that the benefit depends on the exercise intensity.MethodsWe subjected mice to low intensity (Li), moderate intensity (Mi) or high intensity (Hi) exercise, or untrained control (Co) groups based on their individual maximal running capacity.ResultsWe found that exercise intensity played a critical role in tumor control. Only Mi exercise delayed tumor growth and reduced tumor burden, whereas Li or Hi exercise failed to exert similar antitumor effects. While both Li and Mi exercise normalized the tumor vasculature, only Mi exercise increased tumor infiltrated CD8+ T cells, that also displayed enhanced effector function (higher proliferation and expression of CD69, INFγ, GzmB). Moreover, exercise induced an intensity-dependent mobilization of CD8+ T cells into the bloodstream.ConclusionThese findings shed light on the intricate relationship between exercise intensity and cancer, with implications for personalized and optimal exercise prescriptions for tumor control.</p

Metformin reprograms pancreatic stellate cells (PSCS) and tumor-associated macrophages (TAMs), alleviates the fibro-inflammatory tumor microenvironment and reduces metastasis.

<p>Metformin treatment reduces collagen-I and HA production by PSCs, leading to decreased fibrosis in PDACs. Metformin treatment also reduces cytokine production, infiltration and M2 polarization of TAMs, leading to decreased inflammation. This associated with improved desmoplasia and reduced extracellular matrix (ECM) remodeling, epithelial-to-mesenchymal transition (EMT), and metastasis.</p

Metformin reduces collagen-I/hyaluronan production by pancreatic stellate cells.

<p>(A) PSCs were incubated <i>in vitro</i> with metformin (1 mM) for 48h. (A-i) Representative immunocytochemistry images showing the effect of metformin on tumor hyaluronan and collagen-I levels in human pancreatic stellate cells (PSCs) <i>in vitro</i> (n = 2). (A-ii) Quantification of hyaluronan expression in PSCs. Metformin decreases the expression of hyaluronan in PSCs. (A-iii) Quantification of the expression of collagen-I in PSCs. Metformin decreases the expression of collagen-I in PSCs. αSMA denotes activated PSCs. (B) Representative Western blots for the expression of fibrosis-related markers and signaling proteins in PSCs treated with metformin at 0, 0.1, 1 and 10mM. Metformin decreases the expression of fibrosis-related markers and signaling proteins in PSCs. Densitometric analysis of protein expression normalized to ß-actin or total protein (in the case of phosphorylated proteins) is depicted as numbers below the representative bands. Data in A are presented as the mean ± standard error. *p < 0.05 vs. control.</p

Metformin reduces ECM remodeling, EMT, and metastasis in a PDAC mouse model.

<p>(A) Expression of genes associated with extra-cellular matrix (ECM) remodeling, epithelial-to-mesenchymal transition (EMT) and inflammation in AK4.4 tumors from control and metformin-treated mice. Data normalized to control group. 3–4 samples per group pooled in one single PCR array plate. Metformin reduces the expression of pro-tumor genes and increases the expression of anti-tumor genes. (B-i) Representative Western blots showing the effect of metformin (300 mg/Kg) on MMPs and EMT markers in AK4.4 tumors. (B-ii) Densitometric analysis of protein expression normalized to ß-actin. Metformin decreases the expression of MMP-9 and vimentin and increases the expression of e-cadherin in AK4.4 tumors. (C) MMP activity in AK4.4 tumor protein extracts from control and metformin-treated mice (n = 3–4). Metformin decreases the activity of MMPs. (D) Effect of metformin on the percentage of mice affected (incidence) with mesenteric (peritoneal) and abdominal wall (retroperitoneal) metastasis in AK4.4 and PAN02 models (n = 3–8). Metformin reduced the percentage of mice that develop wall metastasis in the more metastatic model (PAN02 model) and induced a tendency for reduced wall as well as mesenteric metastasis in the less metastatic AK4.4 model. (E) Effect of metformin on the number (average) of mesenteric (peritoneal) and abdominal wall (retroperitoneal) metastasis per mouse in the AK4.4 and PAN02 models (n = 3–8). Metformin reduced the number of wall metastasis in the PAN02 model. There were also trends for fewer mesenteric metastasis in AK4.4 and PAN02 tumors. Data in B, C and E are presented as the mean ± standard error. *p < 0.05 vs. control.</p

Metformin treatment associates with reduced hyaluronan levels in human pancreatic cancers in overweight/obese patients.

<p>(i) Representative histology images showing the effect of metformin on tumor hyaluronan levels in normal weight [Body mass index (BMI)<25)] or overweight/obese patients (BMI>25) (n = 22 controls, 7 metformin). (ii) Immunohistochemical analysis of total tumor hyaluronan levels. Metformin decreases the hyaluronan-positive area fraction (%) in overweight/obese patients. Data are presented as the mean ± standard error. * p < 0.05 vs. control in patients with BMI >25.</p

Metformin reprograms TAMs and reduces inflammation in tumors.

<p>(A-i) Representative immunocytochemistry images showing the effect of metformin metformin on the expression of F4/80 (immunofluorescence) in AK4.4 tumors (percentage of viable tumor area) (n = 4). (A-ii) Metformin-treated tumors (300 mg/kg in drinking water) had significantly reduced levels of F4/80-positive tumor-associated macrophages (TAMs). (B) Effect of metformin (0–0.2mM) on the gene expression (qPCR) of M1 (i) and M2 (ii) markers in RAW 264.7 cells (mouse leukaemic monocyte-macrophages) <i>in vitro</i>. Clinically relevant doses (0.05 mM) of metformin treatment reduces expression of M2 markers in macrophages <i>in vitro</i>, including Arg-1 and IL-10. (C) Effect of metformin on the gene expression (qPCR) of M1 (i) and M2 (ii) markers in TAMs isolated from PAN02 tumors <i>in vivo</i> (n = 3). Metformin treatment reduced expression of the M2 markers Arg-1 and IL-10 in TAMs <i>in vivo</i>. (D) Representative Western blots for the expression of signaling proteins in RAW 264.7 cells treated with metformin at 0, 0.05, 0.1, 0.2 and 0.4 mM. Metformin decreases the activation of signaling pathways and increased activation of AMPKα on RAW cells. Densitometric analysis of protein expression normalized to ß-actin or total protein (in the case of phosphorylated protein) is depicted as numbers below the representative bands. (E) Effect of metformin on the protein expression of major cytokines in AK4.4 tumors (n = 4–5) using multiplex protein array. Metformin treatment associated with reduced IL-1ß and CXCL1 expression in tumors. Data are presented as mean ± standard error in A, C and E. * p < 0.05, ** p < 0.01 vs. control.</p

Micelle-Encapsulated Quantum Dot-Porphyrin Assemblies as <i>in Vivo</i> Two-Photon Oxygen Sensors

Micelles have been employed to encapsulate the supramolecular assembly of quantum dots with palladium­(II) porphyrins for the quantification of O₂ levels in aqueous media and <i>in vivo</i>. Förster resonance energy transfer from the quantum dot (QD) to the palladium porphyrin provides a means for signal transduction under both one- and two-photon excitation. The palladium porphyrins are sensitive to O₂ concentrations in the range of 0–160 Torr. The micelle-encapsulated QD-porphyrin assemblies have been employed for <i>in vivo</i> multiphoton imaging and lifetime-based oxygen measurements in mice with chronic dorsal skinfold chambers or cranial windows. Our results establish the utility of the QD-micelle approach for <i>in vivo</i> biological sensing applications