Dataset Information

Non-Invasive In Vivo Imaging and Quantification of Tumor Growth and Metastasis in Rats Using Cells Expressing Far-Red Fluorescence Protein.

ABSTRACT: Non-invasive in vivo imaging is emerging as an important tool for basic and preclinical research. Near-infrared (NIR) fluorescence dyes and probes have been used for non-invasive optical imaging since in the NIR region absorption and auto fluorescence by body tissue is low, thus permitting for greater penetration depths and high signal to noise ratio. Currently, cell tracking systems rely on labeling cells prior to injection or administering probes targeting the cell population of choice right before imaging. These approaches do not enable imaging of tumor growth, as the cell label is diluted during cell division. In this study we have developed cell lines stably expressing the far-red fluorescence protein E2-Crimson, thus enabling continuous detection and quantification of tumor growth. In a xenograft rat model, we show that E2-Crimson expressing cells can be detected over a 5 week period using optical imaging. Fluorescence intensities correlated with tumor volume and weight and allowed for a reliable and robust quantification of the entire tumor compartment. Using a novel injection regime, the seeding of MDA-MB-231 breast cancer cells in the lungs in a rat model was established and verified.

SUBMITTER: Christensen J

PROVIDER: S-EPMC4505884 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Publications

Non-Invasive In Vivo Imaging and Quantification of Tumor Growth and Metastasis in Rats Using Cells Expressing Far-Red Fluorescence Protein.

Christensen Jon J Vonwil Daniel D Shastri V Prasad VP

PloS one 20150717 7

Non-invasive in vivo imaging is emerging as an important tool for basic and preclinical research. Near-infrared (NIR) fluorescence dyes and probes have been used for non-invasive optical imaging since in the NIR region absorption and auto fluorescence by body tissue is low, thus permitting for greater penetration depths and high signal to noise ratio. Currently, cell tracking systems rely on labeling cells prior to injection or administering probes targeting the cell population of choice right b ...[more]

PMID: 26186005

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Project description:BackgroundHuman African trypanosomiasis is caused by infection with parasites of the Trypanosoma brucei species complex, and threatens over 70 million people in sub-Saharan Africa. Development of new drugs is hampered by the limitations of current rodent models, particularly for stage II infections, which occur once parasites have accessed the CNS. Bioluminescence imaging of pathogens expressing firefly luciferase (emission maximum 562 nm) has been adopted in a number of in vivo models of disease to monitor dissemination, drug-treatment and the role of immune responses. However, lack of sensitivity in detecting deep tissue bioluminescence at wavelengths below 600 nm has restricted the wide-spread use of in vivo imaging to investigate infections with T. brucei and other trypanosomatids.Methodology/principal findingsHere, we report a system that allows the detection of fewer than 100 bioluminescent T. brucei parasites in a murine model. As a reporter, we used a codon-optimised red-shifted Photinus pyralis luciferase (PpyRE9H) with a peak emission of 617 nm. Maximal expression was obtained following targeted integration of the gene, flanked by an upstream 5'-variant surface glycoprotein untranslated region (UTR) and a downstream 3'-tubulin UTR, into a T. brucei ribosomal DNA locus. Expression was stable in the absence of selective drug for at least 3 months and was not associated with detectable phenotypic changes. Parasite dissemination and drug efficacy could be monitored in real time, and brain infections were readily detectable. The level of sensitivity in vivo was significantly greater than achievable with a yellow firefly luciferase reporter.Conclusions/significanceThe optimised bioluminescent reporter line described here will significantly enhance the application of in vivo imaging to study stage II African trypanosomiasis in murine models. The greatly increased sensitivity provides a new framework for investigating host-parasite relationships, particularly in the context of CNS infections. It should be ideally suited to drug evaluation programmes.

Project description:Broomrape, caused by the root holoparasite Orobanche cumana, is the main biotic constraint to sunflower oil production worldwide. By the time broomrape emerges, most of the metabolic imbalance has been produced by O. cumana to sunflower plants. UV-induced multicolor fluorescence imaging (MCFI) provides information on the fluorescence emitted by chlorophyll (Chl) a of plants in the spectral bands with peaks near 680 nm (red, F680) and 740 nm (far-red, F740). In this work MCFI was extensively applied to sunflowers, either healthy or parasitized plants, for the first time. The distribution of red and far-red fluorescence was analyzed in healthy sunflower grown in pots under greenhouse conditions. Fluorescence patterns were analyzed across the leaf surface and throughout the plant by comparing the first four leaf pairs (LPs) between the second and fifth week of growth. Similar fluorescence patterns, with a delay of 3 or 4 days between them, were obtained for LPs of healthy sunflower, showing that red and far-red fluorescence varied with the developmental stage of the leaf. The use of F680 and F740 as indicators of sunflower infection by O. cumana during underground development stages of the parasite was also evaluated under similar experimental conditions. Early increases in F680 and F740 as well as decreases in F680/F740 were detected upon infection by O. cumana. Significant differences between inoculated and control plants depended on the LP that was considered at any time. Measurements of Chl contents and final total Chl content supported the results of MCFI, but they were less sensitive in differentiating healthy from inoculated plants. Sunflower infection was confirmed by the presence of broomrape nodules in the roots at the end of the experiment. The potential of MCFI in the red and far-red region for an early detection of O. cumana infection in sunflower was revealed. This technique might have a particular interest for early phenotyping in sunflower breeding programs. To our knowledge, this is the first work where the effect of a parasitic plant in its host is analyzed by means of fluorescence imaging in the red and far-red spectral regions.

Dataset Information

Non-Invasive In Vivo Imaging and Quantification of Tumor Growth and Metastasis in Rats Using Cells Expressing Far-Red Fluorescence Protein.

Publications

Non-Invasive In Vivo Imaging and Quantification of Tumor Growth and Metastasis in Rats Using Cells Expressing Far-Red Fluorescence Protein.

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