Warren D. Foltz

Curcumin Prevents High Fat Diet Induced Insulin Resistance and Obesity via Attenuating Lipogenesis in Liver and Inflammatory Pathway in Adipocytes

Background Mechanisms underlying the attenuation of body weight gain and insulin resistance in response to high fat diet (HFD) by the curry compound curcumin need to be further explored. Although the attenuation of the inflammatory pathway is an accepted mechanism, a recent study suggested that curcumin stimulates Wnt signaling pathway and hence suppresses adipogenic differentiation. This is in contrast with the known repressive effect of curcumin on Wnt signaling in other cell lineages. Methodology and Principal Findings We conducted the examination on low fat diet, or HFD fed C57BL/6J mice with or without curcumin intervention for 28 weeks. Curcumin significantly attenuated the effect of HFD on glucose disposal, body weight/fat gain, as well as the development of insulin resistance. No stimulatory effect on Wnt activation was observed in the mature fat tissue. In addition, curcumin did not stimulate Wnt signaling in vitro in primary rat adipocytes. Furthermore, curcumin inhibited lipogenic gene expression in the liver and blocked the effects of HFD on macrophage infiltration and the inflammatory pathway in the adipose tissue. Conclusions and Significance We conclude that the beneficial effect of curcumin during HFD consumption is mediated by attenuating lipogenic gene expression in the liver and the inflammatory response in the adipose tissue, in the absence of stimulation of Wnt signaling in mature adipocytes.

MRI monitoring of intratumoral drug delivery and prediction of the therapeutic effect with a multifunctional thermosensitive liposome

Non-invasive in vivo imaging of drug distribution enables real-time monitoring and prediction of therapeutic responses to treatment. We have developed a thermosensitive liposomal formulation (HaT: Hyperthermia-activated-cytoToxic) consisting of DPPC and Brij78, a formulation that enhanced drug delivery compared to the lyso-lipid temperature sensitive liposomes (LTSL). Here we report the development of a multifunctional HaT liposome co-encapsulating Gd-DTPA (an MRI probe) and doxorubicin (DOX), which simultaneously releases and reports on drug delivery in a locally heated tumor. The temperature-dependent release profiles of DOX from HaT were closely related to the change in the MR T(1) relaxation time, in which DOX was 100% released at 40-42 °C in 3 min, accompanied by a 60% reduction in T(1). By T(1) relaxometry analysis, no Gd-DTPA leakage was detected in 30 min at 30-37 °C. In the in vivo study, DOX uptake in the tumor was quantitatively correlated with T(1) response (R(2) = 0.98) and the patterns of the T(1) image and the intratumoral DOX uptake were matched, in which both signals were predominantly detected in the highly perfused tumor periphery. Finally, the extent of T(1) relaxation enhancement in the heated tumor successfully predicted the antitumor efficacy in a standard pharmacological response model (R(2) = 0.98).

Optimized spiral imaging for measurement of myocardial T2 relaxation.

Microcirculation oxygen levels and blood volumes should be reflected in measurements of myocardial T2 relaxation. This work describes the optimization of a spiral imaging strategy for robust myocardial T2 measurement to minimize the standard deviation of T2 measurement (σT2). Theoretical and experimental studies of blurring at muscle/blood interfaces enabled the derivation of parameter sets which reduce σ T2 to the level of 5%. T2 relaxation mapping within healthy volunteers provided estimation of residual σT2 within the optimized technique. The standard deviation in T2 measurement across regions of interest (ROIs) in different locations is about 9%. The standard deviation in T2 measurement in an ROI across different time points is about 5%. Magn Reson Med 49:1089–1097, 2003.

Comparing oxygen-sensitive MRI (BOLD R2*) with oxygen electrode measurements: A pilot study in men with prostate cancer

Purpose: To explore the relationship between oxygen-sensitive Magnetic Resonance Imaging (MRI) and oxygen measurements in prostate cancer. Methods: Nine men underwent MRI examinations followed by needle oxygen measurements of tumor bearing region within prostate gland and five men further consented to biopsy. Median pO2 and hypoxic fraction < 5 mm Hg (HP5) were derived. Biopsies were immunostained for Carbonic Anhydrase IX (CA IX), Hypoxia Inducible Factor-1 (HIF 1) and Glucose Transporter-1 (GLUT 1). Corresponding Regions-of-Interest (ROI) were delineated on T2-weighted (T2w) MRI by two observers. Median R2* was calculated for each ROI. Spearman correlation was calculated between R2* and HP5/pO2. Results: MRI quality evaluation resulted in exclusion of 4/18 ROI due to motion (n = 2) and rectal air susceptibility artifact (n = 2). Quality of remaining data was validated by concordance of R2* with T2w, indices and with secondary observer R2* (r = 0.94, p = 0.005). Correlation was observed between R2* and HP5 (r = 0.76, p = 0.02) and a trend was noted between R2* and pO2 (r = −0.66, p = 0.07). GLUT 1 and HIF 1 were expressed in all patients, and CA IX was expressed in one patient with high HP5 (77%) and low pO2 (1.4 mm Hg). Conclusions: MRI using R2* quantification is a promising tool for non-invasive imaging of prostate cancer hypoxia.

Coronary venous oximetry using MRI

Based on the Fick law, coronary venous blood oxygen measurements have value for assessing functional parameters such as the coronary flow reserve. At present, the application of this measure is restricted by its invasive nature. This report describes the design and testing of a noninvasive coronary venous blood oxygen measurement using MRI, with a preliminary focus on the coronary sinus. After design optimization including a four‐coil phased array and an optimal set of data acquisition parameters, quality tests indicate measurement precision on the order of the gold standard optical measurement (3%O2). Comparative studies using catheter sampling suggest reasonable accuracy (3 subjects), with variability dominated by sampling location uncertainty (∼7%O2). Intravenous dipyridamole (5 subjects) induces significant changes in sinus blood oxygenation (22 ± 9% O2), corresponding to flow reserves of 1.8 ± 0.4, suggesting the potential for clinical utility. Underestimation of flow reserve is dominated by right atrial mixing and the systemic effects of dipyridamole. Magn Reson Med 42:837–848, 1999.

Docetaxel Conjugate Nanoparticles That Target α-Smooth Muscle Actin–Expressing Stromal Cells Suppress Breast Cancer Metastasis

Docetaxel-conjugate nanoparticles, known as Cellax, were synthesized by covalently conjugating docetaxel and polyethylene glycol to acetylated carboxymethylcellulose via ester linkages, yielding a polymeric conjugate that self-assembled into 120 nm particles suitable for intravenous administration. In 4T1 and MDA-MB-231 orthotopic breast tumor models, Cellax therapy reduced α-smooth muscle actin (α-SMA) content by 82% and 70%, respectively, whereas native docetaxel and nab-paclitaxel (albumin-paclitaxel nanoparticle, Abraxane) exerted no significant antistromal activity. In Cellax-treated mice, tumor perfusion was increased by approximately 70-fold (FITC-lectin binding), tumor vascular permeability was enhanced by more than 30% (dynamic contrast-enhanced magnetic resonance imaging), tumor matrix was decreased by 2.5-fold (immunohistochemistry), and tumor interstitial fluid pressure was suppressed by approximately 3-fold after Cellax therapy compared with the control, native docetaxel, and nab-paclitaxel groups. The antistromal effect of Cellax treatment corresponded to a significantly enhanced antimetastatic effect: lung nodules were reduced by 7- to 24-fold by Cellax treatment, whereas native docetaxel and nab-paclitaxel treatments were ineffective. Studies of the 4T1 tumor showed that more than 85% of the Cellax nanoparticles were delivered to the α-SMA+ stroma. Significant tumor stromal depletion occurred within 16 hours (∼50% depletion) postinjection, and the α-SMA+ stroma population was almost undetectable (∼3%) by 1 week. The 4T1 tumor epithelial cell population was not significantly reduced in the week after Cellax injection. These data suggest that Cellax targets tumor stroma and performs more efficaciously than docetaxel and nab-paclitaxel.

Tumor-targeted drug delivery using MR-contrasted docetaxel – Carboxymethylcellulose nanoparticles

A carboxymethylcellulose-based polymer conjugate with nanoparticle forming properties (Cellax) has been shown to enhance the pharmacokinetics, specificity of biodistribution, anti-tumor efficacy and safety of docetaxel (DTX) in comparison to the Taxotere™ formulation. We examined Cellax and Taxotere efficacy in four tumor models (EMT-6, B16F10, PC3, and MDA-MB-231), and observed variances in efficacy. To explore whether differences in tumor uptake of Cellax were responsible for these effects, we incorporated superparamagnetic iron oxide nanoparticles (SPIONs) into Cellax particles to enable magnetic resonance (MR) imaging (Cellax-MR). In the EMT-6 tumor model, Cellax-MR nanoparticles exhibited peak tumor accumulation 3-24 h post intravenous injection, and 3 days post-treatment, significant MR contrast was still detected. The amount of Cellax-MR deposited in the EMT-6 tumors was quantifiable as a hypointense volume fraction, a value positively correlated with drug content as determined by LC/MS analysis (R(2) = 0.97). In the four tumor models, Cellax-MR uptake was linearly associated with anti-tumor efficacy (R(2) > 0.9), and was correlated with blood vessel density (R(2) > 0.9). We have affirmed that nanoparticle uptake is variable in tumor physiology, and that this efficacy-predictive parameter can be non-invasively estimated in real-time using a theranostic variant of Cellax.

Surgical ventricular restoration with a cell- and cytokine-seeded biodegradable scaffold

Late after a myocardial infarction (MI), surgical ventricular restoration (SVR) can reduce left ventricular volumes, but an enhanced cardiac patch may be required to restore function. We developed a new, biodegradable patch (modified gelfoam, MGF) consisting of a spongy inner core (gelfoam) to encourage cell engraftment and an outer coating (poly epsilon-caprolactone) to provide sufficient strength to permit ventricular repair. Two weeks after coronary ligation in rats, SVR was performed using one of the following: gelfoam, MGF, MGF patches with hydrogel alone, or with hydrogel and cytokines (stem cell factor, stromal cell-derived factor-1alpha), bone marrow mesenchymal stem cells, or both. Cardiac function and morphology were evaluated by echocardiography, conduction catheterization, magnetic resonance imaging, and histology. Animals whose hearts were repaired with untreated gelfoam died of ventricular rupture. The MGF groups had significantly improved myocardial systolic function vs. MI controls. Enhancement with cytokines and/or cells promoted more alpha-smooth muscle actin-positive cells, more capillaries, greater wall thickness, a more ellipsoid shape, greater fractional shortening, and better-preserved systolic elastance than MGF alone. This combination of the new, reinforced, biodegradable biomaterial and cytokine/cell treatment created a viable tissue after SVR and produced better functional outcomes than un-reinforced gelfoam or MGF alone.

Proximal Cerebral Arteries Develop Myogenic Responsiveness in Heart Failure via Tumor Necrosis Factor-α–Dependent Activation of Sphingosine-1-Phosphate Signaling

Background— Heart failure is associated with neurological deficits, including cognitive dysfunction. However, the molecular mechanisms underlying reduced cerebral blood flow in the early stages of heart failure, particularly when blood pressure is minimally affected, are not known. Methods and Results— Using a myocardial infarction model in mice, we demonstrate a tumor necrosis factor-&agr; (TNF&agr;)–dependent enhancement of posterior cerebral artery tone that reduces cerebral blood flow before any overt changes in brain structure and function. TNF&agr; expression is increased in mouse posterior cerebral artery smooth muscle cells at 6 weeks after myocardial infarction. Coordinately, isolated posterior cerebral arteries display augmented myogenic tone, which can be fully reversed in vitro by the competitive TNF&agr; antagonist etanercept. TNF&agr; mediates its effect via a sphingosine-1-phosphate (S1P)–dependent mechanism, requiring sphingosine kinase 1 and the S1P2 receptor. In vivo, sphingosine kinase 1 deletion prevents and etanercept (2-week treatment initiated 6 weeks after myocardial infarction) reverses the reduction of cerebral blood flow, without improving cardiac function. Conclusions— Cerebral artery vasoconstriction and decreased cerebral blood flow occur early in an animal model of heart failure; these perturbations are reversed by interrupting TNF&agr;/S1P signaling. This signaling pathway may represent a potential therapeutic target to improve cognitive function in heart failure.

Vasodilator Response Assessment in Porcine Myocardium With Magnetic Resonance Relaxometry

Background—This research describes an early preclinical study of the biophysical mechanisms governing changes in myocardial T2 during vasodilation in normal myocardium. Methods and Results—Theoretical modeling and experimental studies in an instrumented pig model (n=7) provided measures of changes in myocardial T2, relative blood volume (BV), and microcirculation oxygen levels (%O2) during intracoronary adenosine infusion. Intracoronary adenosine increases perfusion without increasing blood volume or cardiac metabolic rate; thus, T2 elevations should reflect elevated microcirculation oxygen levels. Robust strategies were used for magnetic resonance imaging (MRI) data collection. Measures of myocardial and vascular T1 before and after Clariscan (Amersham Health) injection provided blood volume assessment. Changes in microcirculation oxygen levels were estimated via direct blood sampling from the left anterior descending (LAD) coronary vein. Perfusion changes were monitored using a Doppler flow wire within the left main coronary artery. Myocardial T2 elevations (&Dgr;T2=17±8%) within the LAD arterial perfusion bed were related to elevations in perfusion (coronary velocity reserve=3.2±0.4) and coronary venous %O2 [&Dgr;(LAD CV%O2) =56±11%], whereas blood volume (&Dgr;BV=0 ±2%) and cardiac metabolic rate [&Dgr;(heart rate x blood pressure) = −4±11%] remained constant. Conclusions—Myocardial T2 elevation during intracoronary adenosine infusion was significant and repeatable, caused by increases in microcirculation oxygen levels. Changes in microcirculation oxygen levels of approximately 40%O2 should be detectable by this technique. This sensitivity should suffice for differentiating normal from abnormal myocardium via measurement of myocardial perfusion reserve.