Real-time in vivo ROS monitoring with luminescent nanoparticles reveals skin inflammation dynamics.

Reactive oxygen species (ROS) are key regulators in numerous pathological contexts, including cancer or inflammation. Their role is complex, which justifies the need for methods enabling their quantitative and time-resolved monitoring in vivo, in the perspective to profile tissues of individual patients. However, current ROS detection methods do not provide these features. Here, we propose a new method based on the imaging of lanthanide-ion nanoparticles (GdVO4:Eu), whose photoluminescence is modulated by the surrounding ROS concentration. We monitored their luminescence after intradermic injection in a mouse ear submitted to an inflammation-inducing topical stimulus. Based on this approach, we quantified the ROS concentration after inflammation induction and identified a two-step kinetics of ROS production, which may be attributed to the response of resident immune cells and their further recruitment at the inflammation locus.

Exploitation of the chick embryo chorioallantoic membrane (CAM) as a platform for anti-metastatic drug testing.

The establishment of clinically relevant models for tumor metastasis and drug testing is a major challenge in cancer research. Here we report a physiologically relevant assay enabling quantitative analysis of metastatic capacity of tumor cells following implantation into the chorioallantoic membrane (CAM). Engraftment of as few as 103 non-small cell lung cancer (NSCLC) and prostate cancer (PCa) cell lines was sufficient for both primary tumor and metastasis formation. Standard 2D-imaging as well as 3D optical tomography imaging were used for the detection of fluorescent metastatic foci in the chick embryo. H2228- and H1975-initiated metastases were confirmed by genomic analysis. We quantified the inhibitory effect of docetaxel on LNCaP, and that of cisplatin on A549- and H1299-initiated metastatic growths. The CAM assay also mimicked the sensitivity of ALK-rearranged H2228 and EGFR-mutated H1975 NSCLC cells to tyrosine kinase inhibitors crizotinib and gefitinib respectively, as well as sensitivity of LNCaP cells to androgen-dependent enzalutamide therapy. The assay was suggested to reconstitute the bone metastatic tropism of PCa cells. We show that the CAM chick embryo model may be a powerful preclinical platform for testing and targeting of the metastatic capacity of cancer cells.

[Validation of a new methodology for characterizing ultrasound contrast agents (UCA)]. / Validation d'un banc hydrodynamique pour la caractérisation de produits de contraste ultrasonores (PCUS).

PURPOSE: Validation of an experimental ultrasound system on erythrocyte suspensions with variable levels of aggregation and application to the echogenicity quantification of UCA under quasi-physiologic flow conditions. MATERIALS AND METHODS. The system is constituted with a Couette cell with variable applied shear rates, an ultrasound emitter/receiver and a digital scope for radio-frequency signal acquisition. Ultrasound indices (UI) were defined for the two experimental established protocols based on the gold standard laser methodology. Washed red cells with or without variable Dextran 70 kD concentrations were used to simulate a wide particle size range. A preliminary application to UCA was conducted with Levovist for calibration of the system. RESULTS: For each protocol, applied ten times on identical whole blood samples, a student t-test revealed no significant variation for all UI. Results on washed red cells were in good agreement with Rayleigh's theory of ultrasound backscattering. Significant correlations were obtained between laser and UI for washed red cells with different Dextran concentrations. An elevation of 12.13 dB in backscattered intensity was obtained after addition of Levovist. CONCLUSION: The constituted Couette system allowed reproducible and accurate echogenicity quantification of small scatterers such as UCA in quasi-physiologic blood flow conditions.

[Functional and early evaluation of treatments in oncology: interest of ultrasonographic contrast agents]. / Evaluation fonctionnelle et précoce des traitements en cancérologie: intérêt des produits de contraste ultrasonores.

The early and functional evaluation of new treatments in oncology is a main goal. At present, technical advances in Doppler-ultrasonography allow the detection of neovascularization for superficial and deep malignant tumors in order to evaluate the efficiency of new treatments as Treatment response can be early predicted according to modifications of this vascularization before Volume modification. Contrast agents injection improves the efficiency of this technique and developments in perfusion softwares optimize this detection.

Dynamic assessment of antiangiogenic therapy by monitoring both tumoral vascularization and tissue degeneration.

Tumor growth is dependent both on endothelial and tumor cells. The aim of this study was to investigate dynamically whether changes in tumor vasculature implicate tumor tissue degeneration during antiangiogenic therapies. In order to quantify intra-tumor vascularization and necrosis, we have used ultrasound technology. This study has identified essential parameters needed to quantify specifically and sensitively the number of microvessels and the extent of necrosis in xenografted human carcinomas during natural tumor evolution, using contrast-enhanced high-frequency ultrasonography with (HFCDUS) or without (HFUS) color Doppler. We showed that quantification of intra-tumor microvessels between HFCDUS and immunohistochemistry is correlated using an anti-CD31 antibody. Furthermore, quantification of tumor necrosis with HFUS was confirmed by histological examination of hematoxylin-eosin-saffranin-stained sections over the observation period. Subsequently, for the assessment of novel angiogenic inhibitors, HFCDUS and HFUS were used to elucidate the underlying dynamics linking vessel inhibition and tumor eradication. We describe a novel application for HFCDUS/HFUS that constitutes an effective, convenient, and non-invasive method for clinical assessment of angiogenic inhibitors. In conclusion, we showed that tumor cells abruptly became necrotic following an antivascular therapy, whereas untreated tumors were protected from degeneration by a significant blood supply.

Full kringles of plasminogen (aa 1-566) mediate complete regression of human MDA-MB-231 breast tumor xenografted in nude mice.

Since kringle (K)5, not present in the angiostatin molecule, was shown to be a key functional domain possessing potent antiangiogenic activity, we have evaluated a new plasminogen-derived fragment, consisting of the N-terminal part of human plasminogen, that included the complete secondary structure of K1-5 (aa 1-566). In contrast to other fragments described to date, K1-5 includes cysteine residues at positions 543, 555 and 560 allowing the formation of the three disulfide bonds lying within K5. Vascular endothelial cell proliferation and migration assays revealed that a replication-defective adenovirus (AdK1-5(1-566)), expressing K1-5 (aa 1-566), was dose dependently more potent that AdK1-3(1-354), an adenovirus that expresses only the first three kringles. In contrast to AdK1-3(1-354), a single intratumoral injection of AdK1-5(1-566) into MDA-MB-231 breast human carcinoma tumors was followed by a total regression of 40% of the tumor and by significant arrest of tumor growth (90%), which was correlated with a drastic decrease of functional neovascularization into the tumors. Furthermore, systemic delivery of AdK1-5(1-566) in mice inhibited the lung invasion of melanoma B16-F10 cells by 87%. Our findings provide evidence that the full kringles of plasminogen (aa 1-566) may be much more potent than K1-3 (aa 1-354), for the suppression of angiogenesis, tumor growth and metastatic dissemination.