3D Raman imaging of systemic endothelial dysfunction in the murine model of metastatic breast cancer

Abstract

It was recently reported in the murine model of metastatic breast cancer (4T1) that tumor progression and development of metastasis is associated with systemic endothelial dysfunction characterized by impaired nitric oxide (NO) production. Using Raman 3D confocal imaging with the analysis of the individual layers of the vascular wall combined with AFM endothelial surface imaging, we demonstrated that metastasis-induced systemic endothelial dysfunction resulted in distinct chemical changes in the endothelium of the aorta. These changes, manifested as a significant increase in the protein content (18 %) and a slight decrease in the lipid content (4 %), were limited to the endothelium and did not occur in the deeper layers of the vascular wall. The altered lipid to protein ratio in the endothelium, although more pronounced in the fixed vascular wall, was also observed in the freshly isolated unfixed vascular wall samples in the aqueous environment (12 and 7 % change of protein and lipid content, respectively). Our results support the finding that the metastasis induces systemic endothelial dysfunction that may contribute to cancer progression.Graphical AbstractSchematic illustration of methodology of sample preparation and measurementElectronic supplementary materialThe online version of this article (doi:10.1007/s00216-016-9436-9) contains supplementary material, which is available to authorized users.

Rapid biochemical profiling of endothelial dysfunction in diabetes, hypertension and cancer metastasis by hierarchical cluster analysis of Raman spectra

“…the 4%‐decrease in the lipid level and the 18%‐increase in the protein content compared with the control. These changes, observed in the endothelium, indicated that it could be a good target in the diagnosis of endothelial dysfunction associated with metastatic breast cancer . To confirm that, HCA was applied to classify the spectra of the endothelium of the control animals and mice with developed cancer metastasis (Fig.…”

“…An important factor for diseases determination is also time, necessary for obtaining the information about the sample and analysis of this information. Encouraged by our previous results related to pathology investigations using Raman imaging in this work, we decided to check if Raman spectroscopy is indeed such a potent diagnostic tool to study the condition of endothelium. Therefore, we extracted 50–70 random, single, recorded in 0.2 s, spectra from the endothelium of mice with diseases and control animals and subjected them to HCA.…”

“…In this way, the set of Raman images is obtained. These images can be assigned to lower and upper part of tissue corresponding to the media layer and endothelium, respectively . From the upper Raman image, where the signal originates from the endothelium layer, the single spectra were extracted and used to HCA.…”

“…The chemical content of endothelium was measured in three murine models of diseases associated with endothelial dysfunction: type 2 diabetes mellitus (T2DM) (db/db mice), [34] hypertension (induced by L-NAME) [33] and metastatic breast cancer. [36,37] Spectral differences mainly connected with alterations in lipid to protein ratio or changes in the secondary structure of proteins enabled successful differentiation of the spectra from healthy mice as compared with mice with endothelial dysfunction with high sensitivity and specificity. Furthermore, our approach proved also useful to differentiate the endothelial dysfunction linked to diabetes from that linked to hypertension or cancer metastasis.…”

Functional inhibition of F11 receptor (F11R/junctional adhesion molecule-A/JAM-A) activity by a F11R-derived peptide in breast cancer and its microenvironment

“…Experiments were performed in vivo in the murine 4T1 breast cancer model [34][35][36]. Plasma samples obtained from the blood of 4T1-treated mice and untreated mice were analyzed by ELISA for soluble F11R/JAM-A (sJAM-A) quantitation.…”