A novel 3D high-content assay identifies compounds that prevent fibroblast invasion into tissue surrogates

Wenzel, Carsten; Otto, Saskia A.; Prechtl, Stefan; Parczyk, Karsten; Steigemann, Patrick

doi:10.1016/j.yexcr.2015.10.003

Cited by 22 publications

(14 citation statements)

References 45 publications

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“…Although spheroid size and shape remain the primary parameters for real-time monitoring of a drug effect ( 21 , 38 , 39 ), the use of fluorescent probes provides additional information regarding the mechanism of drug action ( 20 , 23 , 47 , 48 ). In principle, the more probes you use, the more information you gather.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, many reports have focused on the difference in drug cytotoxicity in various cancer models ( 46 , 74 , 105 – 107 ). Beyond the cytotoxicity issue, 3D models have also been applied to discover molecules that suppress tumor-relevant processes, such as hypoxia-induced drug resistance ( 23 ), metastasis-like invasion and cancer cell spreading ( 21 ), and cancer-associated fibroblast infiltration ( 20 ). Often, analysis of complex phenotypes requires high-content monitoring of multiple parameters.…”

Section: Discussionmentioning

confidence: 99%

“…Many techniques have been developed to generate spheroids ( 12 , 24 ), including growing cells in spinner flasks ( 25 ), in hanging drops ( 26 ), in levitation ( 27 ), or microgravity ( 28 ) within a natural ( 29 , 30 ) or synthetic ( 31 ) polymer matrix and in liquid-overlay culture on agar- ( 32 ), agarose- ( 22 ), or polyHEMA-coated ( 33 , 34 ) plates. In particular, agarose coating of standard microtiter plates creates concave-bottomed wells used to produce an array of individual spheroids and subsequent large-scale drug screening in a 3D format ( 20 , 22 , 23 ). However, because of reproducibility issues, in-lab coating is not always suitable for automated spheroid imaging ( 21 ).…”

Section: Introductionmentioning

confidence: 99%

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High-Content Monitoring of Drug Effects in a 3D Spheroid Model

et al. 2017

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A recent decline in the discovery of novel medications challenges the widespread use of 2D monolayer cell assays in the drug discovery process. As a result, the need for more appropriate cellular models of human physiology and disease has renewed the interest in spheroid 3D culture as a pertinent model for drug screening. However, despite technological progress that has significantly simplified spheroid production and analysis, the seeming complexity of the 3D approach has delayed its adoption in many laboratories. The present report demonstrates that the use of a spheroid model may be straightforward and can provide information that is not directly available with a standard 2D approach. We describe a cost-efficient method that allows for the production of an array of uniform spheroids, their staining with vital dyes, real-time monitoring of drug effects, and an ATP-endpoint assay, all in the same 96-well U-bottom plate. To demonstrate the method performance, we analyzed the effect of the preclinical anticancer drug MLN4924 on spheroids formed by VCaP and LNCaP prostate cancer cells. The drug has different outcomes in these cell lines, varying from cell cycle arrest and protective dormancy to senescence and apoptosis. We demonstrate that by using high-content analysis of spheroid arrays, the effect of the drug can be described as a series of EC50 values that clearly dissect the cytostatic and cytotoxic drug actions. The method was further evaluated using four standard cancer chemotherapeutics with different mechanisms of action, and the effect of each drug is described as a unique multi-EC50 diagram. Once fully validated in a wider range of conditions, this method could be particularly valuable for phenotype-based drug discovery.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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High-Content Monitoring of Drug Effects in a 3D Spheroid Model

et al. 2017

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“…However, differentiated iPSC assays, combined with more informative functional screening technology, provide improved models of normal and diseased tissue compared with traditional assays, leading many investigators to anticipate they will ultimately improve clinical success rates. Further high-content image-based 3D spheroid assays have been applied to smallmolecule screens investigating compounds that specifically target dormant tumour cells within the inner core of tumour spheroids or compounds which prevent either fibroblast or immune invasion into tumour spheroids [114][115][116] . Such 3D spheroid assays formats have also been used as tissue surrogates to study immune infiltration into specific tissue types and represent a rapid and cost-effective alternative to animal models for studying hostimmune response 114,115 .…”

Section: Developments To Improve Disease Modelling [Au:ok?]mentioning

confidence: 99%

Screening out irrelevant cell-based models of disease

Horváth

Aulner

Bickle

et al. 2016

Nat Rev Drug Discov

426

321

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The common and persistent failures to translate promising preclinical drug candidates into clinical success highlight the limited effectiveness of disease models currently used in drug discovery. An apparent reluctance to explore and adopt alternative cell- and tissue-based model systems, coupled with a detachment from clinical practice during assay validation, contributes to ineffective translational research. To help address these issues and stimulate debate, here we propose a set of principles to facilitate the definition and development of disease-relevant assays, and we discuss new opportunities for exploiting the latest advances in cell-based assay technologies in drug discovery, including induced pluripotent stem cells, three-dimensional (3D) co-culture and organ-on-a-chip systems, complemented by advances in single-cell imaging and gene editing technologies. Funding to support precompetitive, multidisciplinary collaborations to develop novel preclinical models and cell-based screening technologies could have a key role in improving their clinical relevance, and ultimately increase clinical success rates.

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“…For example, the importance of the latter in the progression to invasive carcinoma was demonstrated by Bischel et al, who showed that ductal carcinoma in situ cells patterned in a luminal structure progressed to an invasive phenotype in the presence of fibroblasts . In another example, Bayer Pharma AG used arrays of co‐culture spheroids consisting of either MCF7 cells or H1299 cells coated in fluorescently labeled human pulmonary fibroblasts to perform a high‐content phenotypic screen of 480 known bioactive substances and identified both prostaglandins and Rho‐associated protein kinase (ROCK) inhibitors as compounds that inhibited fibroblast invasion. Paper‐supported multilayer models have also been used to explore the effect of stromal cells and an oxygen gradient on local invasion.…”

Section: Current Progress In Modeling Cancer Biology In Vitromentioning

confidence: 99%

The Current Landscape of 3D In Vitro Tumor Models: What Cancer Hallmarks Are Accessible for Drug Discovery?

Rodenhizer

Dean

D’Arcangelo

et al. 2018

Adv Healthcare Materials

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Cancer prognosis remains a lottery dependent on cancer type, disease stage at diagnosis, and personal genetics. While investment in research is at an all-time high, new drugs are more likely to fail in clinical trials today than in the 1970s. In this review, a summary of current survival statistics in North America is provided, followed by an overview of the modern drug discovery process, classes of models used throughout different stages, and challenges associated with drug development efficiency are highlighted. Then, an overview of the cancer hallmarks that drive clinical progression is provided, and the range of available clinical therapies within the context of these hallmarks is categorized. Specifically, it is found that historically, the development of therapies is limited to a subset of possible targets. This provides evidence for the opportunities offered by novel disease-relevant in vitro models that enable identification of novel targets that facilitate interactions between the tumor cells and their surrounding microenvironment. Next, an overview of the models currently reported in literature is provided, and the cancer biology they have been used to explore is highlighted. Finally, four priority areas are suggested for the field to accelerate adoption of in vitro tumour models for cancer drug discovery.

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A novel 3D high-content assay identifies compounds that prevent fibroblast invasion into tissue surrogates

Cited by 22 publications

References 45 publications

High-Content Monitoring of Drug Effects in a 3D Spheroid Model

High-Content Monitoring of Drug Effects in a 3D Spheroid Model

Screening out irrelevant cell-based models of disease

The Current Landscape of 3D In Vitro Tumor Models: What Cancer Hallmarks Are Accessible for Drug Discovery?

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