Dataset Information

A Novel 3D Model for Visualization and Tracking of Fibroblast-Guided Directional Cancer Cell Migration.

ABSTRACT: Stromal fibroblasts surrounding cancer cells are a major and important constituent of the tumor microenvironment not least because they contain cancer-associated fibroblasts, a unique fibroblastic cell type that promotes tumorigenicity through extracellular matrix remodeling and secretion of soluble factors that stimulate cell differentiation and invasion. Despite much progress made in understanding the molecular mechanisms that underpin fibroblast-tumor cross-talk, relatively little is known about the way the two cell types interact from a physical contact perspective. In this study, we report a novel three-dimensional dumbbell model that would allow the physical interaction between the fibroblasts and cancer cells to be visualized and monitored by microscopy. To achieve the effect, the fibroblasts and cancer cells in 50% Matrigel suspension were seeded as independent droplets in separation from each other. To allow for cell migration and interaction, a narrow passage of Matrigel causeway was constructed in between the droplets, effectively molding the gel into the shape of a dumbbell. Under time-lapse microscopy, we were able to visualize and image the entire process of fibroblast-guided cancer cell migration event, from initial vessel-like structure formation by the fibroblasts to their subsequent invasion across the causeway, attracting and trapping the cancer cells in the process. Upon prolonged culture, the entire population of fibroblasts eventually infiltrated across the passage and condensed into a spheroid-like cell mass, encapsulating the bulk of the cancer cell population within. Suitable for almost every cell type, our model has the potential for a wider application as it can be adapted for use in drug screening and the study of cellular factors involved in cell-cell attraction.

SUBMITTER: Zhang Y

PROVIDER: S-EPMC7600195 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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A Novel 3D Model for Visualization and Tracking of Fibroblast-Guided Directional Cancer Cell Migration.

Zhang Yihe Y Jiang Bingjie B Lee Meng Huee MH

Biology 20201008 10

Stromal fibroblasts surrounding cancer cells are a major and important constituent of the tumor microenvironment not least because they contain cancer-associated fibroblasts, a unique fibroblastic cell type that promotes tumorigenicity through extracellular matrix remodeling and secretion of soluble factors that stimulate cell differentiation and invasion. Despite much progress made in understanding the molecular mechanisms that underpin fibroblast-tumor cross-talk, relatively little is known ab ...[more]

PMID: 33049958

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Project description:Inversin is a ciliary protein that critically regulates developmental processes and tissue homeostasis in vertebrates, partly through the degradation of Dishevelled (Dvl) proteins to coordinate Wnt signaling in planar cell polarity (PCP). Here, we investigated the role of Inversin in coordinating cell migration, which highly depends on polarity processes at the single-cell level, including the spatial and temporal organization of the cytoskeleton as well as expression and cellular localization of proteins in leading edge formation of migrating cells. Using cultures of mouse embryonic fibroblasts (MEFs) derived from inv(-/-) and inv(+/+) animals, we confirmed that both inv(-/-) and inv(+/+) MEFs form primary cilia, and that Inversin localizes to the primary cilium in inv(+/+) MEFs. In wound healing assays, inv(-/-) MEFs were severely compromised in their migratory ability and exhibited cytoskeletal rearrangements, including distorted lamellipodia formation and cilia orientation. Transcriptome analysis revealed dysregulation of Wnt signaling and of pathways regulating actin organization and focal adhesions in inv(-/-) MEFs as compared to inv(+/+) MEFs. Further, Dvl-1 and Dvl-3 localized to MEF primary cilia, and β-catenin/Wnt signaling was elevated in inv(-/-) MEFs, which moreover showed reduced ciliary localization of Dvl-3. Finally, inv(-/-) MEFs displayed dramatically altered activity and localization of RhoA, Rac1, and Cdc42 GTPases, and aberrant expression and targeting of the Na(+)/H(+) exchanger NHE1 and ezrin/radixin/moesin (ERM) proteins to the edge of cells facing the wound. Phosphorylation of β-catenin at the ciliary base and formation of well-defined lamellipodia with localization and activation of ERM to the leading edge of migrating cells were restored in inv(-/-) MEFs expressing Inv-GFP. Collectively, our findings point to the significance of Inversin in controlling cell migration processes, at least in part through transcriptional regulation of genes involved in Wnt signaling and pathways that control cytoskeletal organization and ion transport.

Project description:IntroductionCardiovascular magnetic resonance (CMR)-guided cardiac catheterization is becoming more widespread due to the ability to acquire both functional CMR measurements and diagnostic catheterization data without exposing patients to ionizing radiation. However, the real-time imaging sequences used for catheter guidance during these procedures are limited in resolution and the anatomical detail they can provide. In this study, we propose a passive catheter tracking approach which simultaneously improves catheter tracking and visualization of the anatomy.Methods60 patients with congenital heart disease underwent CMR-guided cardiac catheterization on a 1.5T CMR scanner (Ingenia, Philips Healthcare, Best the Netherlands) using the Philips iSuite system. The proposed T1-overlay technique uses a commercially available heavily T1-weighted sequence to image the catheter, and overlays it on a high-resolution 3D dataset within iSuite in real-time. Suppressed tissue in the real-time images enables the use of a thick imaging slab to assist in tracking of the catheter. Improvement in catheter visualization time was compared between T1-overlay and the conventional invasive CMR (iCMR) balanced steady state free precession (bSSFP) sequence. This technique also enabled selective angiography visualization for real-time evaluation of blood flow dynamics (such as pulmonary transit time), similar to direct contrast injection under standard fluoroscopy. Estimates of pulmonary transit time using iCMR were validated using x-ray fluoroscopy in 16 patients.ResultsThe T1-overlay approach significantly increased the time that the catheter tip was kept in view by the technologist compared to the bSSFP sequence conventionally used for iCMR. The resulting images received higher ratings for blood/balloon contrast, anatomy visualization, and overall suitability for iCMR guidance by three cardiologists. iCMR selective angiography using T1-overlay also provided accurate estimates of pulmonary transit time that agreed well with x-ray fluoroscopy.ConclusionWe demonstrate a new passive catheter tracking technique using the iSuite platform that improves visualization of the catheter and cardiac anatomy. These improvements significantly increase the time that the catheter tip is seen throughout the procedure. We also demonstrate the feasibility of iCMR selective angiography for the measurement of pulmonary transit time.

Dataset Information

A Novel 3D Model for Visualization and Tracking of Fibroblast-Guided Directional Cancer Cell Migration.

Publications

A Novel 3D Model for Visualization and Tracking of Fibroblast-Guided Directional Cancer Cell Migration.

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