Imaging Circulating Tumor Cells in Freely Moving Awake Small Animals Using a Miniaturized Intravital Microscope

Sasportas, Laura S.; Gambhir, Sanjiv S.

doi:10.1371/journal.pone.0086759

Cited by 34 publications

(30 citation statements)

References 32 publications

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“…This has been confirmed by Juratli et al who observed high variations in CTC numbers in consecutive IVFC 5-min imaging windows and in sequentially drawn patient blood samples [45]. We envision that this problem may be addressed in the future using continuous intravital imaging methods in freely moving small animals [34].…”

Section: Discussionsupporting

confidence: 59%

“…Using two rounds of fluorescence activated cell sorting (FACS), we established a stable cell line (denoted 4T1-GL) that expresses high levels of the bifusion reporter gene for at least 12 passages in culture [34]. Using an additional round of FACS, we selected the 4T1-GL-TS population as the 5% brightest cells among 4T1-GL cells, based on GFP fluorescence.…”

Section: Methodsmentioning

confidence: 99%

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Detection and Quantitation of Circulating Tumor Cell Dynamics by Bioluminescence Imaging in an Orthotopic Mammary Carcinoma Model

et al. 2014

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Circulating tumor cells (CTCs) have been detected in the bloodstream of both early-stage and advanced cancer patients. However, very little is know about the dynamics of CTCs during cancer progression and the clinical relevance of longitudinal CTC enumeration. To address this, we developed a simple bioluminescence imaging assay to detect CTCs in mouse models of metastasis. In a 4T1 orthotopic metastatic mammary carcinoma mouse model, we demonstrated that this quantitative method offers sensitivity down to 2 CTCs in 0.1–1mL blood samples and high specificity for CTCs originating from the primary tumor, independently of their epithelial status. In this model, we simultaneously monitored blood CTC dynamics, primary tumor growth, and lung metastasis progression over the course of 24 days. Early in tumor development, we observed low numbers of CTCs in blood samples (10–15 cells/100 µL) and demonstrated that CTC dynamics correlate with viable primary tumor growth. To our knowledge, these data represent the first reported use of bioluminescence imaging to detect CTCs and quantify their dynamics in any cancer mouse model. This new assay is opening the door to the study of CTC dynamics in a variety of animal models. These studies may inform clinical decision on the appropriate timing of blood sampling and value of longitudinal CTC enumeration in cancer patients.

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Section: Discussionsupporting

confidence: 59%

Section: Methodsmentioning

confidence: 99%

Detection and Quantitation of Circulating Tumor Cell Dynamics by Bioluminescence Imaging in an Orthotopic Mammary Carcinoma Model

et al. 2014

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“…S9), suggesting the potential for reinitiation of cluster transit after occlusion and/or a slow "creeping" transit behavior. This observation may explain the reportedly short circulating half-lives of CTCs (37) and CTC clusters (9) because increased residence times within capillaries may inhibit/delay their detection or sampling from larger vessels. Further studies are needed to explore these phenomena.…”

Section: Resultsmentioning

confidence: 99%

Clusters of circulating tumor cells traverse capillary-sized vessels

Storey

Moore

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

414

356

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Multicellular aggregates of circulating tumor cells (CTC clusters) are potent initiators of distant organ metastasis. However, it is currently assumed that CTC clusters are too large to pass through narrow vessels to reach these organs. Here, we present evidence that challenges this assumption through the use of microfluidic devices designed to mimic human capillary constrictions and CTC clusters obtained from patient and cancer cell origins. Over 90% of clusters containing up to 20 cells successfully traversed 5-to 10-μm constrictions even in whole blood. Clusters rapidly and reversibly reorganized into single-file chain-like geometries that substantially reduced their hydrodynamic resistances. Xenotransplantation of human CTC clusters into zebrafish showed similar reorganization and transit through capillary-sized vessels in vivo. Preliminary experiments demonstrated that clusters could be disrupted during transit using drugs that affected cellular interaction energies. These findings suggest that CTC clusters may contribute a greater role to tumor dissemination than previously believed and may point to strategies for combating CTC cluster-initiated metastasis.microfluidics | cancer metastasis | CTC clusters | circulating tumor cell cluster microemboli | capillary microhemodynamics

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“…For all bioluminescence work, we used genetically-modified GBM2, GBM39 and U87-MG whereby cells were transfected with lentiviral vectors that expressed either firefly luciferase (for GBM39) or a fusion protein of GFP and firefly luciferase (for GBM2 and U87) [22, 23]. All BLI work done on an IVIS Spectrum (Xenogen Corporation, Alameda, CA).…”

Section: Methodsmentioning

confidence: 99%

Synergistic inhibition of glioma cell proliferation by Withaferin A and tumor treating fields

et al. 2017

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BACKGROUND Glioblastoma (GBM) is the most aggressive and lethal form of brain cancer. Standard therapies are non-specific and often of limited effectiveness; thus, efforts are underway to uncover novel, unorthodox therapies against GBM. In previous studies, we investigated Withaferin A, a steroidal lactone from Ayurvedic medicine that inhibits proliferation in cancers including GBM. Another novel approach, tumor treating fields (TTFields), is thought to disrupt mitotic spindle formation and stymie proliferation of actively dividing cells. We hypothesized that combining TTFields with Withaferin A would synergistically inhibit proliferation in glioblastoma. METHODS Human glioblastoma cells (GBM2, GBM39, U87-MG) and human breast adenocarcinoma cells (MDA-MB-231) were isolated from primary tumors. The glioma cell lines were genetically engineered to express firefly luciferase. Proliferative potential was assessed either by bioluminescence imaging or cell counting via hemocytometer. RESULTS TTFields (4 V/cm) significantly inhibited growth of the four cancer cell lines tested (n=3 experiments per time point, 4 measurements per sample, p<0.02 at least; 2-way ANOVA, control vs. treatment). The combination of Withaferin A (10–100 nM) with TTFields significantly inhibited the growth of the glioma cells to a degree beyond that of Withaferin A or TTFields alone. The interaction of the Withaferin A and TTFields on glioma cells was found to be synergistic in nature (p<0.01, n=3 experiments). These findings were validated by both bioluminescence and hemocytometric measurements. CONCLUSIONS The combination of Withaferin A with TTFields represents a novel approach to treat GBM in a manner that is likely better than either treatment alone and that is synergistic.

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Imaging Circulating Tumor Cells in Freely Moving Awake Small Animals Using a Miniaturized Intravital Microscope

Cited by 34 publications

References 32 publications

Detection and Quantitation of Circulating Tumor Cell Dynamics by Bioluminescence Imaging in an Orthotopic Mammary Carcinoma Model

Detection and Quantitation of Circulating Tumor Cell Dynamics by Bioluminescence Imaging in an Orthotopic Mammary Carcinoma Model

Clusters of circulating tumor cells traverse capillary-sized vessels

Synergistic inhibition of glioma cell proliferation by Withaferin A and tumor treating fields

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