HTS approaches to voltage-gated ion channel drug discovery

Denyer, Jane; Worley, Jennings F.; Cox, Brian J.; Allenby, Gary; Banks, Martyn

doi:10.1016/s1359-6446(98)01199-4

Cited by 94 publications

(58 citation statements)

References 20 publications

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“…There is tremendous interest in the pharmaceutical industry for improved high-throughput cell-based screening platforms to expedite target validation as well as for use in preclinical trials (Sundberg, 2000). Currently, cellbased drug screening assays have been implemented for numerous molecular interactions, including ion channel activity (Denyer et al, 1998;Gonzalez et al, 1999), Gprotein-coupled receptor response (Conway et al, 1999), and induction of apoptosis (Lee et al, 2003). Multiplexed devices such as microtiter plates and array bioreactors have been developed for large-scale screening applications (Kostov et al, 2001;Maffia et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays

Hung

Lee

Sabounchi

et al. 2004

Biotechnol. Bioeng.

475

345

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Section: Introductionmentioning

confidence: 99%

Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays

Hung

Lee

Sabounchi

et al. 2004

Biotechnol. Bioeng.

475

345

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“…VGCCs have been an emerging important drug target, as the dysfunction of VGCCs has proved to be related to diseases in the cardiovascular, central nervous, endocrine, and other systems. 39 With the predominant adoption of fluorescence-based detection in cell-based screening programs, the present study was carried out with a calcium fluorescent indicator combined with laser scanning confocal microscopy. Figure 4 shows confocal images of 2D cells on PLLA flat substrates (A), the overlay of the confocal fluorescence on light microscopic images for N2D (B) and 3D (C) cells on microwell patterns after Calcium Green-1 loading, as well as a typical plot for changes in cytoplasmic Calcium Green-1 fluorescence intensity for both a responding and non-responding SH-SY5Y cell upon 50 mM high K + depolarization (D).…”

Section: Vgcc Responsiveness Of Sh-sy5y Cells In Microwell Patterns Vmentioning

confidence: 99%

Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)

Wang²,

Zhang³

et al. 2013

IJN

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Introduction:In this study, quasi-three-dimensional (3D) microwell patterns were fabricated with poly (l-lactic acid) for the development of cell-based assays, targeting voltage-gated calcium channels (VGCCs). Methods and materials: SH-SY5Y human neuroblastoma cells were interfaced with the microwell patterns and found to grow as two dimensional (2D), 3D, and near two dimensional (N2D), categorized on the basis of the cells' location in the pattern. The capability of the microwell patterns to support 3D cell growth was evaluated in terms of the percentage of the cells in each growth category. Cell spreading was analyzed in terms of projection areas under light microscopy. SH-SY5Y cells' VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium Green TM -1. The expression of L-type calcium channels was evaluated using immunofluorescence staining with DM-BODIPY. Results: It was found that cells within the microwells, either N2D or 3D, showed more rounded shapes and less projection areas than 2D cells on flat poly (l-lactic acid) substrates. Also, cells in microwells showed a significantly lower VGCC responsiveness than cells on flat substrates, in terms of both response magnitudes and percentages of responsive cells, upon depolarization with 50 mM K + . This lower VGCC responsiveness could not be explained by the difference in L-type calcium channel expression. For the two patterns addressed in this study, N2D cells consistently exhibited an intermediate value of either projection areas or VGCC responsiveness between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness. Conclusion: These results suggest that the pattern structure and therefore the cell growth characteristics were critical factors in determining cell VGCC responsiveness and thus provide an approach for engineering cell functionality in cell-based assay systems and tissue engineering scaffolds.

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“…In addition, because of the need to develop subtype-selective compounds, multiple cell lines, expressing ion channels that are closely related to the primary target, must be available for discrete assessment of channel selectivity. Historically, binding assays, whereby novel chemical entities are screened for their ability to displace a radiolabeled ligand specific to the site of interest on the channel have not proven to be reliable in predicting ion channel function (Denyer et al, 1998). In addition, by virtue of the novelty of the interactions that are being targeted, specific radioligands have been difficult to develop.…”

Section: Drug Development For Ion Channel Modulators: Hts Limitationsmentioning

confidence: 99%

Drugability of Extracellular Targets: Discovery of Small Molecule Drugs Targeting Allosteric, Functional, and Subunit-Selective Sites on GPCRs and Ion Channels

Grigoriadis

Hoare

Lechner

et al. 2008

Neuropsychopharmacol

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Beginning with the discovery of the structure of deoxyribose nucleic acid in 1953, by James Watson and Francis Crick, the sequencing of the entire human genome some 50 years later, has begun to quantify the classes and types of proteins that may have relevance to human disease with the promise of rapidly identifying compounds that can modulate these proteins so as to have a beneficial and therapeutic outcome. This so called 'drugable space' involves a variety of membrane-bound proteins including the superfamily of G-protein-coupled receptors (GPCRs), ion channels, and transporters among others. The recent number of novel therapeutics targeting membrane-bound extracellular proteins that have reached the market in the past 20 years however pales in magnitude when compared, during the same timeframe, to the advancements made in the technologies available to aid in the discovery of these novel therapeutics. This review will consider select examples of extracellular drugable targets and focus on the GPCRs and ion channels highlighting the corticotropin releasing factor (CRF) type 1 and g-aminobutyric acid receptors, and the Ca V 2.2 voltage-gated ion channel. These examples will elaborate current technological advancements in drug discovery and provide a prospective framework for future drug development.

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HTS approaches to voltage-gated ion channel drug discovery

Cited by 94 publications

References 20 publications

Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays

Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays

Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)

Drugability of Extracellular Targets: Discovery of Small Molecule Drugs Targeting Allosteric, Functional, and Subunit-Selective Sites on GPCRs and Ion Channels

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