Short-term circulating tumor cell dynamics in mouse xenograft models and implications for liquid biopsy

Williams, Amber; Fitzgerald, Jessica E.; Ivich, Fernando; Sontag, Eduardo D.; Niedre, Mark

doi:10.1101/814368

Cited by 5 publications

(17 citation statements)

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“…We also sincerely thank Dr. Miguel Martin (Universidad Complutense, Madrid, Spain) for kindly sharing data. This manuscript has been released as a pre-print at ( 51 ).…”

Section: Acknowledgmentsmentioning

confidence: 99%

Short-Term Circulating Tumor Cell Dynamics in Mouse Xenograft Models and Implications for Liquid Biopsy

et al. 2020

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Motivation: Circulating tumor cells (CTCs) are widely studied using liquid biopsy methods that analyze fractionally-small peripheral blood (PB) samples. However, little is known about natural fluctuations in CTC numbers that may occur over short timescales in vivo, and how these may affect detection and enumeration of rare CTCs from small blood samples. Methods: We recently developed an optical instrument called "diffuse in vivo flow cytometry" (DiFC) that uniquely allows continuous, non-invasive counting of rare, green fluorescent protein expressing CTCs in large blood vessels in mice. Here, we used DiFC to study short-term changes in CTC numbers in multiple myeloma and Lewis lung carcinoma xenograft models. We analyzed CTC detections in over 100 h of DiFC data, and considered intervals corresponding to approximately 1%, 5%, 10%, and 20% of the PB volume. In addition, we analyzed changes in CTC numbers over 24 h (diurnal) periods. Results: For rare CTCs (fewer than 1 CTC per ml of blood), the use of short DiFC intervals (corresponding to small PB samples) frequently resulted in no detections. For more abundant CTCs, CTC numbers frequently varied by an order of magnitude or more over the timescales considered. This variance in CTC detections far exceeded that expected by Poisson statistics or by instrument variability. Rather, the data were consistent with significant changes in mean numbers of CTCs on the timescales of minutes and hours. Conclusions: The observed temporal changes can be explained by known properties of CTCs, namely, the continuous shedding of CTCs from tumors and the short half-life of CTCs in blood. It follows that the number of cells in a blood sample are strongly impacted by the timing of the draw. The issue is likely to be compounded for multicellular CTC clusters or specific CTC subtypes, which are even more rare than single CTCs. However, we show that enumeration can in principle be improved by averaging multiple samples, analysis of larger volumes, or development of methods for enumeration of CTCs directly in vivo.

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“…We also sincerely thank Dr. Miguel Martin (Universidad Complutense, Madrid, Spain) for kindly sharing data. This manuscript has been released as a pre-print at ( 51 ).…”

Section: Acknowledgmentsmentioning

confidence: 99%

Short-Term Circulating Tumor Cell Dynamics in Mouse Xenograft Models and Implications for Liquid Biopsy

et al. 2020

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“…Previous studies have reported the underlying mechanisms of the kinetics of CTCs (binding/unbinding mechanism), adhesion processes, , their survival mechanisms in blood, and interactions with blood cells and endothelium, using diverse set of ligands and receptors . Various in vivo animal models , and in vitro models for CTC adhesion assays − and their intravastion/extravasation have accordingly been explored previously. Despite these advancements, dynamic evolution of CTCs in a physiologically replicative vascular model is yet to be deciphered.…”

Section: Introductionmentioning

confidence: 99%

Frugal Approach toward Developing a Biomimetic, Microfluidic Network-on-a-Chip for In Vitro Analysis of Microvascular Physiology

Priyadarshani

Roy

Das

et al. 2021

ACS Biomater. Sci. Eng.

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Several disease conditions, such as cancer metastasis and atherosclerosis, are deeply connected with the complex biophysical phenomena taking place in the complicated architecture of the tiny blood vessels in human circulatory systems. Traditionally, these diseases have been probed by devising various animal models, which are otherwise constrained by ethical considerations as well as limited predictive capabilities. Development of an engineered network-on-a-chip, which replicates not only the functional aspects of the blood-carrying microvessels of human bodies, but also its geometrical complexity and hierarchical microstructure, is therefore central to the evaluation of organ-assist devices and disease models for therapeutic assessment. Overcoming the constraints of reported resource-intensive fabrication techniques, here, we report a facile, simple yet niche combination of surface engineering and microfabrication strategy to devise a highly ordered hierarchical microtubular network embedded within a polydimethylsiloxane (PDMS) slab for dynamic cell culture on a chip, with a vision of addressing the exclusive aspects of the vascular transport processes under medically relevant paradigms. The design consists of hierarchical complexity ranging from capillaries (∼80 μm) to large arteries (∼390 μm) and a simultaneous tuning of the interfacial material chemistry. The fluid flow behavior is characterized numerically within the hierarchical network, and a confluent endothelial layer is realized on the inner wall of microfluidic device. We further explore the efficacy of the device as a vascular deposition assay of circulatory tumor cells (MG-63 osteosarcoma cells) present in whole blood. The proposed paradigm of mimicking an in vitro vascular network in a low-cost paradigm holds further potential for probing cellular dynamics as well as offering critical insights into various vascular transport processes.

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“…Although these are widely used in biomedical research, they are far from optimal for a number of reasons. In particular, small blood samples provide poor statistical sampling of the circulating blood volume [13][14][15], blood is known to degrade rapidly after removal from the body [16], and enrichment can cause cell loss or dissolution [17][18]. Moreover, in the case of small animal studies CTCs may be so rare that it is necessary to draw and analyze the entire blood volume, which requires euthanizing the animal [19].…”

Section: Introductionmentioning

confidence: 99%

Fluorescence Labeling of Circulating Tumor Cells with Folate Receptor Targeted Molecular Probes for DiffuseIn VivoFlow Cytometry

Patil¹,

Srinivasarao

Amiji

et al. 2020

Preprint

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Purpose: We recently developed a new instrument called 'diffuse in vivo flow cytometry' (DiFC) for enumeration of rare fluorescently-labeled circulating tumor cells (CTCs) in small animals without drawing blood samples. Until now, we have used cell lines that express fluorescent proteins, or were pre-labeled with a fluorescent dye ex-vivo. In this work, we investigated the use of two folate receptor (FR)-targeted fluorescence molecular probes for in vivo labeling of FR+ CTCs for DiFC.Methods: We used EC-17 and Cy5-PEG-FR fluorescent probes. We studied the affinity of these probes for L1210A and KB cancer cells, both of which over-express FR. We tested the labeling specificity in cells in culture in vitro, in whole blood, and in mice in vivo. We also studied detectability of labeled cells with DiFC.Results: Both EC-17 and Cy5-PEG-FR probes had high affinity for FR+ CTCs in cell culture in vitro.However, only EC-17 had sufficient specificity for CTCs in whole blood. EC-17 labeled CTCs were also readily detectable in circulation in mice with DiFC.Conclusions: This work demonstrates the feasibility of labeling CTCs for DiFC with a cell surface receptor targeted probe, greatly expanding the utility of the method for pre-clinical animal models. Because DiFC uses diffuse light, this method could be also used to enumerate CTCs in larger animal models and potentially even in humans.

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Short-term circulating tumor cell dynamics in mouse xenograft models and implications for liquid biopsy

Cited by 5 publications

References 44 publications

Short-Term Circulating Tumor Cell Dynamics in Mouse Xenograft Models and Implications for Liquid Biopsy

Short-Term Circulating Tumor Cell Dynamics in Mouse Xenograft Models and Implications for Liquid Biopsy

Frugal Approach toward Developing a Biomimetic, Microfluidic Network-on-a-Chip for In Vitro Analysis of Microvascular Physiology

Fluorescence Labeling of Circulating Tumor Cells with Folate Receptor Targeted Molecular Probes for DiffuseIn VivoFlow Cytometry

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