scholarly journals Metabolic Switching of Tumor Cells under Hypoxic Conditions in a Tumor-on-a-chip Model

Micromachines ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 382 ◽  
Author(s):  
Valentina Palacio-Castañeda ◽  
Lucas Kooijman ◽  
Bastien Venzac ◽  
Wouter Verdurmen ◽  
Séverine Le Gac
Keyword(s):  
Cell Culture ◽  
Tumor Microenvironment ◽  
Tumor Cell ◽  
Tumor Cells ◽  
In Vitro Models ◽  
Cancer Therapies ◽  
Metabolic Switch ◽  
Hypoxic Conditions ◽  
Tumor Cell Culture

Hypoxia switches the metabolism of tumor cells and induces drug resistance. Currently, no therapeutic exists that effectively and specifically targets hypoxic cells in tumors. Development of such therapeutics critically depends on the availability of in vitro models that accurately recapitulate hypoxia as found in the tumor microenvironment. Here, we report on the design and validation of an easy-to-fabricate tumor-on-a-chip microfluidic platform that robustly emulates the hypoxic tumor microenvironment. The tumor-on-a-chip model consists of a central chamber for 3D tumor cell culture and two side channels for medium perfusion. The microfluidic device is fabricated from polydimethylsiloxane (PDMS), and oxygen diffusion in the device is blocked by an embedded sheet of polymethyl methacrylate (PMMA). Hypoxia was confirmed using oxygen-sensitive probes and the effect on the 3D tumor cell culture investigated by a pH-sensitive dual-labeled fluorescent dextran and a fluorescently labeled glucose analogue. In contrast to control devices without PMMA, PMMA-containing devices gave rise to decreases in oxygen and pH levels as well as an increased consumption of glucose after two days of culture, indicating a rapid metabolic switch of the tumor cells under hypoxic conditions towards increased glycolysis. This platform will open new avenues for testing anti-cancer therapies targeting hypoxic areas.

Influence of Tumor Cell Culture Supernatants on Macrophage Functional Polarization: In Vitro Models of Macrophage-tumor Environment Interaction

2011 ◽  
Vol 97 (5) ◽  
pp. 647-654 ◽  
Author(s):  
Iuliana Caras ◽  
Catalin Tucureanu ◽  
Lucian Lerescu ◽  
Ramona Pitica ◽  
Laura Melinceanu ◽  
...  
Keyword(s):  
Cell Culture ◽  
Tumor Cell ◽  
In Vitro Models ◽  
Environment Interaction ◽  
Tumor Environment ◽  
Tumor Cell Culture ◽  
Functional Polarization ◽  
Culture Supernatants

Comparison between clinical response and in vitro drug sensitivity of primary human tumors in the adhesive tumor cell culture system.

1987 ◽  
Vol 5 (12) ◽  
pp. 1912-1921 ◽  
Author(s):  
J A Ajani ◽  
F L Baker ◽  
G Spitzer ◽  
A Kelly ◽  
W Brock ◽  
...  
Keyword(s):  
Cell Culture ◽  
Tumor Cell ◽  
Culture System ◽  
Distinct Advantage ◽  
Cell Culture System ◽  
True Value ◽  
Tumor Cell Culture ◽  
Prospective Trials ◽  
Tumor Types

The newly described adhesive tumor cell culture system (ATCCS) offers a distinct advantage over other assays in that it has a high plating efficiency requiring low cell inoculum, it affords workable assays in approximately 70% of specimens from the heterogenous tumor types, and it has the ability to assay up to nine drugs at four different concentrations. Clinical correlations based on the ATCCS were obtained in 65 patients undergoing 71 clinical trials. Patients with melanoma, lung cancer, and sarcoma dominated the group. The most active in vitro drug was correlated per clinical trial. Thirteen of 17 (76%) sensitive in vitro predictions and 51 of 54 (94%) resistant in vitro predictions were accurate. The assay in this study had a sensitivity of 81% and specificity of 93%. These preliminary results are encouraging and warrant prospective trials to establish the true value of this assay to patients.

scholarly journals Cancer-Associated Fibroblasts Modulate Transcriptional Signatures Involved in Proliferation, Differentiation and Metastasis in Head and Neck Squamous Cell Carcinoma

Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3361
Author(s):  
Emilia Wiechec ◽  
Mustafa Magan ◽  
Natasa Matic ◽  
Anna Ansell-Schultz ◽  
Matti Kankainen ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Tumor Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
In Vitro Models ◽  
Cancer Associated Fibroblasts ◽  
Mesenchymal Transition ◽  
2D And 3D

Cancer-associated fibroblasts (CAFs) are known to increase tumor growth and to stimulate invasion and metastasis. Increasing evidence suggests that CAFs mediate response to various treatments. HNSCC cell lines were co-cultured with their patient-matched CAFs in 2D and 3D in vitro models, and the tumor cell gene expression profiles were investigated by cDNA microarray and qRT-PCR. The mRNA expression of eight candidate genes was examined in tumor biopsies from 32 HNSCC patients and in five biopsies from normal oral tissue. Differences in overall survival (OS) were tested with Kaplan–Meier long-rank analysis. Thirteen protein coding genes were found to be differentially expressed in tumor cells co-cultured with CAFs in 2D and 81 in 3D when compared to tumor cells cultured without CAFs. Six of these genes were upregulated both in 2D and 3D (POSTN, GREM1, BGN, COL1A2, COL6A3, and COL1A1). Moreover, two genes upregulated in 3D, MMP9 and FMOD, were significantly associated with the OS. In conclusion, we demonstrated in vitro that CAF-derived signals alter the tumor cell expression of multiple genes, several of which are associated with differentiation, epithelial-to-mesenchymal transition (EMT) phenotype, and metastasis. Moreover, six of the most highly upregulated genes were found to be overexpressed in tumor tissue compared to normal tissue.

scholarly journals Rapamycin Promotes ROS-Mediated Cell Death via Functional Inhibition of xCT Expression in Melanoma Under γ-Irradiation

2021 ◽  
Vol 11 ◽  
Author(s):  
Yunseo Woo ◽  
Hyo-Ji Lee ◽  
Jeongyeon Kim ◽  
Seung Goo Kang ◽  
Sungjin Moon ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
Tumor Cells ◽  
Mtor Signaling ◽  
Γ Irradiation ◽  
Γ Radiation ◽  
Hypoxic Conditions ◽  
Tumor Tissues ◽  
Γ Rays

Although many cancer patients are administered radiotherapy for their treatment, the interaction between tumor cells and macrophages in the tumor microenvironment attenuates the curative effects of radiotherapy. The enhanced activation of mTOR signaling in the tumors promotes tumor radioresistance. In this study, the effects of rapamycin on the interaction between tumor cells and macrophages were investigated. Rapamycin and 3BDO were used to regulate the mTOR pathway. In vitro, tumor cells cocultured with macrophages in the presence of each drug under normoxic or hypoxic conditions were irradiated with γ–rays. In vivo, mice were irradiated with γ–radiation after injection with DMSO, rapamycin and 3BDO into tumoral regions. Rapamycin reduced the secretion of IL-4 in tumor cells as well as YM1 in macrophages. Mouse recombinant YM1 decreased the enhanced level of ROS and the colocalized proportion of both xCT and EEA1 in irradiated tumor cells. Human recombinant YKL39 also induced results similar to those of YM1. Moreover, the colocalized proportion of both xCT and LC3 in tumor tissues was elevated by the injection of rapamycin into tumoral regions. Overall, the suppression of mTOR signaling in the tumor microenvironment might be useful for the improvement of tumor radioresistance.

scholarly journals A Hybrid in Silico and Tumor-on-a-Chip Approach to Model Targeted Protein Behavior in 3D Microenvironments

Cancers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2461
Author(s):  
Valentina Palacio-Castañeda ◽  
Simon Dumas ◽  
Philipp Albrecht ◽  
Thijmen J. Wijgers ◽  
Stéphanie Descroix ◽  
...  
Keyword(s):  
Mathematical Modeling ◽  
Tumor Cell ◽  
Tumor Cells ◽  
In Silico ◽  
Growth Factor Receptor ◽  
In Vitro Models ◽  
Protein Activity ◽  
Apparent Diffusion Coefficients ◽  
Ankyrin Repeat Proteins

To rationally improve targeted drug delivery to tumor cells, new methods combining in silico and physiologically relevant in vitro models are needed. This study combines mathematical modeling with 3D in vitro co-culture models to study the delivery of engineered proteins, called designed ankyrin repeat proteins (DARPins), in biomimetic tumor microenvironments containing fibroblasts and tumor cells overexpressing epithelial cell adhesion molecule (EpCAM) or human epithelial growth factor receptor (HER2). In multicellular tumor spheroids, we observed strong binding-site barriers in combination with low apparent diffusion coefficients of 1 µm2·s-1 and 2 µm2 ·s-1 for EpCAM- and HER2-binding DARPin, respectively. Contrasting this, in a tumor-on-a-chip model for investigating delivery in real-time, transport was characterized by hindered diffusion as a consequence of the lower local tumor cell density. Finally, simulations of the diffusion of an EpCAM-targeting DARPin fused to a fragment of Pseudomonas aeruginosa exotoxin A, which specifically kills tumor cells while leaving fibroblasts untouched, correctly predicted the need for concentrations of 10 nM or higher for extensive tumor cell killing on-chip, whereas in 2D models picomolar concentrations were sufficient. These results illustrate the power of combining in vitro models with mathematical modeling to study and predict the protein activity in complex 3D models.

scholarly journals A computational diffusion model to study antibody transport within reconstructed tumor microenvironments

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Ana Luísa Cartaxo ◽  
Jaime Almeida ◽  
Emilio J. Gualda ◽  
Maria Marsal ◽  
Pablo Loza-Alvarez ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
Tumor Cells ◽  
Fluorescent Antibody ◽  
Light Sheet ◽  
In Vitro Models ◽  
Experimental Models ◽  
Computational Framework ◽  
Heterogeneous Distribution ◽  
Alginate Capsules

Abstract Background Antibodies revolutionized cancer treatment over the past decades. Despite their successfully application, there are still challenges to overcome to improve efficacy, such as the heterogeneous distribution of antibodies within tumors. Tumor microenvironment features, such as the distribution of tumor and other cell types and the composition of the extracellular matrix may work together to hinder antibodies from reaching the target tumor cells. To understand these interactions, we propose a framework combining in vitro and in silico models. We took advantage of in vitro cancer models previously developed by our group, consisting of tumor cells and fibroblasts co-cultured in 3D within alginate capsules, for reconstruction of tumor microenvironment features. Results In this work, an experimental-computational framework of antibody transport within alginate capsules was established, assuming a purely diffusive transport, combined with an exponential saturation effect that mimics the saturation of binding sites on the cell surface. Our tumor microenvironment in vitro models were challenged with a fluorescent antibody and its transport recorded using light sheet fluorescence microscopy. Diffusion and saturation parameters of the computational model were adjusted to reproduce the experimental antibody distribution, with root mean square error under 5%. This computational framework is flexible and can simulate different random distributions of tumor microenvironment elements (fibroblasts, cancer cells and collagen fibers) within the capsule. The random distribution algorithm can be tuned to follow the general patterns observed in the experimental models. Conclusions We present a computational and microscopy framework to track and simulate antibody transport within the tumor microenvironment that complements the previously established in vitro models platform. This framework paves the way to the development of a valuable tool to study the influence of different components of the tumor microenvironment on antibody transport.

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