Abstract LB-A22: Comparison of the effect of pharmaceutical compounds on tumor cells in 2D and 3D in vitro models using label-free, quantitative 4 dimensional holographic imaging

e15166 Background: Inhibition of CYP17 by abiraterone acetate (AA) blocks A biosynthesis, reduces A levels and results in inhibition of A-dependent tumor growth. AA has been shown to improve survival in patients with metastatic castrate-resistant prostate cancer (HR = 0.74) (Scher, ASCO 2011). AA is converted to ABI in vivo. This study investigates whether direct effects of ABI on tumor cells (ie, blockade of intratumoral A synthesis) may contribute to the clinical efficacy of AA using in vitro models. Methods: LNCaP cells (human prostate cancer cell line [HPCCL]) were grown in 2D and 3D cultures, and ABI activity was assessed by reduction of androgen receptor (AR) output in gene expression profiling (Affymetrix microarray), reporter assay, and by immunoblot for A-responsive signals. Using ABI-sensitive LNCaP cells, responses were further characterized in the presence or absence of ligand (dihydrotestosterone [DHT]) and with A-targeting agents TOK-001 and MDV3100. Results: ABI exposure resulted in dose-dependent modulation of A-dependent gene expression in Affymetrix microarray analysis of 2D and 3D cultures. Similar results were seen in an analysis of AR-dependent protein and in an AR-driven reporter assay in 2D LNCaP or VCaP (another HPCCL) cell culture. In LNCaP, AR output was inhibited by low concentrations of ABI (IC50 100-300 nM), comparable to TOK-001 (IC50 30-100 nM) and less potent than MDV3100 (IC50 10-30 nM). Growth inhibition (MTT assay) occurred at concentrations 10-fold higher for all compounds. For ABI and TOK-001, inhibition of AR output was reduced by DHT (30- to 100-fold increase in IC50); with MDV3100, the shift in IC50 was less (10-fold) with agonism of AR output at low nM concentrations. Conclusions: ABI has an inhibitory effect on AR output in LNCaP cells. Partially overcoming this inhibition through addition of DHT implies that CYP17 inhibition and the subsequent A reduction play a role in the intratumoral activity of AA. However, persistent inhibition of AR output in the presence of DHT implies additional anti-A effects of AA in prostate tumor cells. This model may provide a useful tool for the study of AR-targeted therapies, combinations of agents, and resistance to agents targeting A.

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

Cancers ◽

10.3390/cancers13133361 ◽

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.

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Effect of lipopeptide structure on gene delivery system properties: Evaluation in 2D and 3D in vitro models

Colloids and Surfaces B Biointerfaces ◽

10.1016/j.colsurfb.2018.04.003 ◽

2018 ◽

Vol 167 ◽

pp. 328-336 ◽

Cited By ~ 7

Author(s):

O.O. Koloskova ◽

A.M. Gileva ◽

M.G. Drozdova ◽

M.V. Grechihina ◽

N.E. Suzina ◽

...

Keyword(s):

Gene Delivery ◽

Delivery System ◽

In Vitro Models ◽

Gene Delivery System ◽

System Properties ◽

2D And 3D

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Chitosan biopolymer: Alternative adhesion factor and scaffold matrix for 2D and 3D neuronal cultures

Biomedical Science and Engineering ◽

10.4081/bse.2019.107 ◽

2020 ◽

Author(s):

Donatella Di Lisa ◽

Mariateresa Tedesco ◽

Elena Dellacasa ◽

Mattia Pesce ◽

Tiziano Catelani ◽

...

Keyword(s):

Cell Cultures ◽

Culture Media ◽

In Vitro Models ◽

Neuronal Cultures ◽

Culture Techniques ◽

Chitosan Biopolymer ◽

2D And 3D ◽

Adhesion Factor

The increase of different types of cell cultures, which can be used for the in vitro studies of physiological and/or pathological processes, has introduced the need to improve culture techniques through the use of materials and culture media that promote growth, recreating a cellular micro-environment that can be asserted in in vivo condition. The standard methods for the functionalization of supports used for cell cultures are based on the use of synthetic or natural biopolymers, which generally have high costs, such as poly-lysine and polyornithine. The aim of this work is to demonstrate the alternative use of the polysaccharide chitosan as adhesion factor and structural component for 2D/3D neuronal cultures. Thanks to its versatility, it could be easily functionalized for the fabrication of personalized of in vitro models

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Encapsulated multicellular tumor spheroids as a novel in vitro model to study small size tumors

Hemijska industrija ◽

10.2298/hemind0312585m ◽

2003 ◽

Vol 57 (12) ◽

pp. 585-588 ◽

Cited By ~ 4

Author(s):

Elena Markvicheva ◽

Lina Bezdetnaya ◽

Artur Bartkowiak ◽

Annie Marc ◽

Jean-Louis Gorgen ◽

...

Keyword(s):

Tumor Cells ◽

Tumor Biology ◽

Cell Encapsulation ◽

Large Cell ◽

In Vitro Models ◽

Multicellular Tumor Spheroids ◽

Tumor Spheroids ◽

Vitro Model

Presently multicellular tumor spheroids (MTS) are being widely used in various aspects of tumor biology, including studies in biology and photodynamic therapy. The cellular organization of spheroids allows the recreation of in vivo small tumors much better than all common two-dimensional in vitro models. The cell encapsulation method could be proposed as a novel technique to quickly and easily prepare a large number of spheroids with narrow size distribution within a desirable diameter range. Moreover, the proposed technique for spheroid generation using encapsulated growing tumor cells could provide entirely new avenues to develop a novel spheroid co-culture model (for instance, the in vitro co-cultvation of tumor cells and monocytes, or epithelial cells, or fibroblasts etc). The current research was aimed at developing a simple and reliable method to encapsulate tumor cells and to cultivate them in vitro. In order to generate spheroids, MCF-7 cells were encapsulated and cultivated in 200 ml T-flasks in a 5% CO2 atmosphere at 37?C for 4-5 weeks. The cell proliferation was easily observed using a light microscope. The cells grew in aggregates increasing in size with time. The cell growth resulted in the formation of large cell clusters (spheroids) which filled the whole microcapsule volume in 4-5 weeks.

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Polysaccharide Multilayer Films in Sensors for Detecting Prostate Tumor Cells Based on Hyaluronan-CD44 Interactions

Cells ◽

10.3390/cells9061563 ◽

2020 ◽

Vol 9 (6) ◽

pp. 1563 ◽

Cited By ~ 1

Author(s):

João Batista Maia Rocha Neto ◽

Andrey Coatrini Soares ◽

Rogério Aparecido Bataglioli ◽

Olívia Carr ◽

Carlos Alberto Rodrigues Costa ◽

...

Keyword(s):

Tumor Cells ◽

Point Of Care ◽

Prostate Tumor ◽

Layer By Layer ◽

Label Free ◽

Point Of Care Diagnostics ◽

Lbl Films ◽

Prostatic Tumor ◽

Prostate Tumor Cells

The increasing need for point-of-care diagnosis has sparked the development of label-free sensing platforms, some of which are based on impedance measurements with biological cells. Here, interdigitated electrodes were functionalized with layer-by-layer (LbL) films of hyaluronan (HA) and chitosan (CHI) to detect prostatic tumor cells (PC3 line). The deposition of LbL films was confirmed with atomic force microscopy and polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), which featured the vibrational modes of the HA top layer capable of interacting specifically with glycoprotein CD44 receptors overexpressed in tumor cells. Though the CHI/HA LbL films cannot be considered as a traditional biosensor due to their limited selectivity, it was possible to distinguish prostate tumor cells in the range from 50 to 600 cells/µL in in vitro experiments with impedance spectroscopy. This was achieved by treating the impedance data with information visualization methods, which confirmed the distinguishing ability of the films by observing the absence of false positives in a series of control experiments. The CD44–HA interactions may, therefore, be exploited in clinical analyses and point-of-care diagnostics for cancer, particularly if computational methods are used to process the data.

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Abstract B38: Ras clipping by bacterial toxin RRSP reduces viability and proliferation of Ras-dependent cancer cell lines in 2D and 3D in vitro models

10.1158/1557-3125.ras18-b38 ◽

2020 ◽

Author(s):

Vania Vidimar ◽

Minyoung Park ◽

Roman A. Melnyk ◽

Karla J.F. Satchell

Keyword(s):

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

In Vitro Models ◽

Bacterial Toxin ◽

2D And 3D

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An Innovative Cell Microincubator for Drug Discovery Based on 3D Silicon Structures

Journal of Nanomaterials ◽

10.1155/2016/8236539 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

Francesca Aredia ◽

Francesca Carpignano ◽

Salvatore Surdo ◽

Giuseppe Barillaro ◽

Giuliano Mazzini ◽

...

Keyword(s):

Drug Discovery ◽

Tumor Cells ◽

Three Dimensional ◽

Label Free ◽

Core Element ◽

Great Ability ◽

Label Free Detection ◽

Ht1080 Cells ◽

Power Radiation

We recently employed three-dimensional (3D) silicon microstructures (SMSs) consisting in arrays of 3 μm-thick silicon walls separated by 50 μm-deep, 5 μm-wide gaps, as microincubators for monitoring the biomechanical properties of tumor cells. They were here applied to investigate the in vitro behavior of HT1080 human fibrosarcoma cells driven to apoptosis by the chemotherapeutic drug Bleomycin. Our results, obtained by fluorescence microscopy, demonstrated that HT1080 cells exhibited a great ability to colonize the narrow gaps. Remarkably, HT1080 cells grown on 3D-SMS, when treated with the DNA damaging agent Bleomycin under conditions leading to apoptosis, tended to shrink, reducing their volume and mimicking the normal behavior of apoptotic cells, and were prone to leave the gaps. Finally, we performed label-free detection of cells adherent to the vertical silicon wall, inside the gap of 3D-SMS, by exploiting optical low coherence reflectometry using infrared, low power radiation. This kind of approach may become a new tool for increasing automation in the drug discovery area. Our results open new perspectives in view of future applications of the 3D-SMS as the core element of a lab-on-a-chip suitable for screening the effect of new molecules potentially able to kill tumor cells.

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3D Bone Morphology Alters Gene Expression, Motility, and Drug Responses in Bone Metastatic Tumor Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21186913 ◽

2020 ◽

Vol 21 (18) ◽

pp. 6913

Author(s):

Ushashi C. Dadwal ◽

Alyssa R. Merkel ◽

Jonathan M. Page ◽

Kristin A. Kwakwa ◽

Michael Kessler ◽

...

Keyword(s):

Gene Expression ◽

Tumor Cells ◽

Metastatic Tumor ◽

Skeletal Metastases ◽

Bone Morphology ◽

3D Scaffolds ◽

Β Receptor ◽

2D And 3D

Patients with advanced skeletal metastases arising from primary cancers including breast, lung, and prostate suffer from extreme pain, bone loss, and frequent fractures. While the importance of interactions between bone and tumors is well-established, our understanding of complex cell–cell and cell–microenvironment interactions remains limited in part due to a lack of appropriate 3D bone models. To improve our understanding of the influence of bone morphometric properties on the regulation of tumor-induced bone disease (TIBD), we utilized bone-like 3D scaffolds in vitro and in vivo. Scaffolds were seeded with tumor cells, and changes in cell motility, proliferation, and gene expression were measured. Genes associated with TIBD significantly increased with increasing scaffold rigidity. Drug response differed when tumors were cultured in 3D compared to 2D. Inhibitors for Integrin β3 and TGF-β Receptor II significantly reduced bone-metastatic gene expression in 2D but not 3D, while treatment with the Gli antagonist GANT58 significantly reduced gene expression in both 2D and 3D. When tumor-seeded 3D scaffolds were implanted into mice, infiltration of myeloid progenitors changed in response to pore size and rigidity. This study demonstrates a versatile 3D model of bone used to study the influence of mechanical and morphometric properties of bone on TIBD.

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