Drug Discovery Approaches Utilizing Three-Dimensional Cell Culture

Ryan, Sarah-Louise; Baird, Anne‐Marie; Vaz, Gisela M. F.; Urquhart, Aaron; Senge, Ma­thias O.; Richard, Derek J.; O’Byrne, Kenneth J.; Davies, Anthony M.

doi:10.1089/adt.2015.670

Cited by 89 publications

(77 citation statements)

References 79 publications

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“…It is well established for cells of various origins [1–3, 37–39], including medulloblastoma [25, 26], that cells cultured in 2D differ from those in 3D cultures. We compared the expression profiles of three differentiation and stem cell markers in ONS-76 cells grown in 2D monolayers and in 3D hydrogels with MAX8 and RGDS-tagged MAX8 cell constructs (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Beta-hairpin hydrogels as scaffolds for high-throughput drug discovery in three-dimensional cell culture

Worthington

Drake

et al. 2017

Analytical Biochemistry

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Automated cell-based high-throughput screening (HTS) is a powerful tool in drug discovery, and it is increasingly being recognized that three-dimensional (3D) models, which more closely mimic in vivo-like conditions, are desirable screening platforms. One limitation hampering the development of 3D HTS is the lack of suitable 3D culture scaffolds that can readily be incorporated into existing HTS infrastructure. We now show that β-hairpin peptide hydrogels can serve as a 3D cell culture platform that is compatible with HTS. MAX8 β-hairpin peptides can physically assemble into a hydrogel with defined porosity, permeability and mechanical stability with encapsulated cells. Most importantly, the hydrogels can then be injected under shear-flow and immediately reheal into a hydrogel with the same properties exhibited prior to injection. The post-injection hydrogels are cell culture compatible at physiological conditions. Using standard HTS equipment and medulloblastoma pediatric brain tumor cells as a model system, we show that automatic distribution of cell-peptide mixtures into 384-well assay plates results in evenly dispensed, viable MAX8-cell constructs suitable for commercially available cell viability assays. Since MAX8 peptides can be functionalized to mimic the microenvironment of cells from a variety of origins, MAX8 peptide gels should have broad applicability for 3D HTS drug discovery.

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Section: Resultsmentioning

confidence: 99%

“…Thus far, there are several different approaches to creating a 3D HTS method, but each comes with its own set of challenges and difficulties [1, 37–39]. The best studied methods employ spheroids or hydrogels.…”

Section: Discussionmentioning

confidence: 99%

Beta-hairpin hydrogels as scaffolds for high-throughput drug discovery in three-dimensional cell culture

Worthington

Drake

et al. 2017

Analytical Biochemistry

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“…Current model systems primarily rely on the monolayer cell cultures and animal models. Simplistic monolayer cultures have their advantages, but they are often significantly different in gene expression, epigenetics, and cell function compared to native 3D tissues . Also, they often lack cell–cell and cell–matrix interactions, leading to the absence of tissue specific properties.…”

Section: Introductionmentioning

confidence: 99%

“…Simplistic monolayer cultures have their advantages, but they are often significantly different in gene expression, epigenetics, and cell function compared to native 3D tissues . Also, they often lack cell–cell and cell–matrix interactions, leading to the absence of tissue specific properties. Although animal models are widely used in biomedical research, they fail to faithfully predict human responses in a physiologically relevant manner due to the significant species divergences .…”

Section: Introductionmentioning

confidence: 99%

Advances in Hydrogels in Organoids and Organs‐on‐a‐Chip

et al. 2019

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The major motivation is to better mimic human physiology and functions at multiscales from the molecular to the cellular, tissue, organ or even whole organism level. Current model systems primarily rely on the monolayer cell cultures and animal models. Simplistic monolayer cultures have their advantages, but they are often significantly different in gene expression, epigenetics, and cell function compared to native 3D tissues. [1,2] Also, they often lack cell-cell and cell-matrix interactions, [2,3] leading to the absence of tissue specific properties. Although animal models are widely used in biomedical research, they fail to faithfully predict human responses in a physiologically relevant manner due to the significant species divergences. [4] The limitations of these systems have provided an impetus for the development of alternative cell-based 3D models in vitro that better resemble the complex functionalities of living organs. Over the last several years, organoids and organ-on-a-chip (OOC), representing the major technological breakthrough, have emerged as two distinct model systems to achieve the same goal of building 3D organotypic models in vitro by bridging the gap between animal models and monolayer cultures. These engineered models are different in terms of cell source, tissue composition, architectural variability, functional features, scales, cellular fidelity, etc. Organoids, primarily evolving from developmental principles, refer to 3D multicellular tissues by self-organization of stem cells or organ-specific progenitors, which can recapitulate the intricate architectures and functionalities of in vivo organs. [5][6][7] Recent breakthroughs in organoid technology have enabled the successful generation of a variety of human organoids, such as the brain, [8,9] intestine, [10,11] liver, [12] kidney, [13,14] lung, [15] etc. These near physiological 3D organoids have garnered momentum for their potential applications in human organ development and disease modeling, drug screening and regenerative medicine. The explosion of organoids research has been catalyzed by the significant progress of stem cell biology and availability of human stem cells. In contrast, the OOC, relying on the bioengineering design principle, is a miniaturized in vitro model that can recreate the functional units of living organs on a microfluidic cell culture device using predifferentiated cells, often cell lines. [1,16] The OOC is primarily evolved from the convergence of tissue engineering and microfabricated technologies, which is characterized by the capability to simulate cellular microenvironment by precise control over fluid flow, mechanical forces and biochemical factors. [4,17] Remarkable progress has been made Significant advances in materials, microscale technology, and stem cell biology have enabled the construction of 3D tissues and organs, which will ultimately lead to more effective diagnostics and therapy. Organoids and organs-on-a-chip (OOC), evolved from developmental biology and bioengineering principles, have e...

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“…The 3D cell-based experiments mimic tumors in some aspects and hence can be used as preliminary investigations before in vivo studies[34–36]. The hanging drop method of obtaining spheroids is advantageous because of its simplicity and reproducibility; moreover, these spheroids resemble tissue[37] unlike in other methods such as extracellular matrix (ECM) scaffolds and hydrogel systems.…”

Section: Resultsmentioning

confidence: 99%

Design of a doxorubicin-peptidomimetic conjugate that targets HER2-positive cancer cells

Pallerla

Gauthier

Sable

et al. 2017

European Journal of Medicinal Chemistry

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Doxorubicin (DOX) belongs to the anthracycline class of drugs that are used in the treatment of various cancers. It has limited cystostatic effects in therapeutic doses, but higher doses can cause cardiotoxicity. In the current approach, we conjugated a peptidomimetic (Arg-aminonaphthylpropionic acid-Phe, compound 5) known to bind to HER2 protein to DOX via a glutaric anhydride linker. Antiproliferative assays suggest that the DOX-peptidomimetic conjugate has activity in the lower micromolar range. The conjugate exhibited higher toxicity in HER2-overexpressed cells than in MCF-7 and MCF-10A cells that do not overexpress HER2 protein. Cellular uptake studies using confocal microscope experiments showed that the conjugate binds to HER2-overexpressed cells and DOX is taken up into the cells in 4 h compared to conjugate in MCF-7 cells. Binding studies using surface plasmon resonance indicated that the conjugate binds to the HER2 extracellular domain with high affinity compared to compound 5 or DOX alone. The conjugate was stable in the presence of cells with a half-life of nearly 4 h and 1 h in human serum. DOX is released from the conjugate and internalized into the cells in 4 h, causing cellular toxicity. These results suggest that this conjugate can be used to target DOX to HER2-overexpressing cells and can improve the therapeutic index of DOX for HER2-positive cancer.

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Drug Discovery Approaches Utilizing Three-Dimensional Cell Culture

Cited by 89 publications

References 79 publications

Beta-hairpin hydrogels as scaffolds for high-throughput drug discovery in three-dimensional cell culture

Beta-hairpin hydrogels as scaffolds for high-throughput drug discovery in three-dimensional cell culture

Advances in Hydrogels in Organoids and Organs‐on‐a‐Chip

Design of a doxorubicin-peptidomimetic conjugate that targets HER2-positive cancer cells

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