Assay and Drug Development Technologies - 5

REVIEWS
Advances in High Content Screening
for Drug Discovery
Kenneth A. Giuliano, Jeffrey R. Haskins, and D. Lansing Taylor
Cellomics, Inc., Pittsburgh, Pennsylvania.
ABSTRACT
Cell-based target validation, secondary screening, lead optimization,
and structure-activity relationships have been recast with the
advent of HCS. Prior to HCS, a computational approach to the
characterization of the functions of specific target proteins and
other cellular constituents, along with whole-cell functions employing
fluorescence cell-based assays and microscopy, required
extensive interaction among the researcher, instrumentation, and
software tools. Early HCS platforms were instrument-centric and
addressed the need to interface fully automated fluorescence
microscopy, plate-handling automation, and seamless image
analysis. HCS has since evolved into an integrated solution for
accelerated drug discovery by encompassing the workflow components
of assay and reagent design, robust instrumentation for
automated fixed-end-point and live cell kinetic analysis, generalized
and specific BioApplication software (Cellomics, Pittsburgh,
PA) modules that produce information on drug responses from cell
image data, and informatics/bioinformatics solutions that build
knowledge from this information while providing a means to
globalize HCS throughout an entire organization. This review
communicates how these recent advances are incorporated into
the drug discovery workflow by presenting a real-world use case.
HCS HAS REVOLUTIONIZED SECONDARY
SCREENING AND LEAD OPTIMIZATION
A
dramatically new approach to early drug discovery
was introduced with the launch of HCS around the
initial ArrayScan (Cellomics, Pittsburgh, PA) instrument
in 1997.1 HCS is a technology platform
designed to define the temporal and spatial activities ofgenes,
proteins, and other cellular constituents in living cells. An
integrated platform must provide instruments, general and
specific BioApplication software (Cellomics), informatics, and
reagents to automatically determine changes in distribution,
environment, and/or activity of fluorescently labeled reporter
molecules in arrays of cells. An automated, computerized
Reprinted with permission from Assay Drug Dev Technol 2003;1(4):565-557.
method for processing, analyzing, displaying, and storing the
data is a key component of the platform that allows analyses
of large numbers of experimental treatments including small
molecules and biologicals that could affect specific cell
functions and constituents.
Although HCS was introduced as a solution for early drug
discovery just a few years ago, several reviews, authored by
researchers within the pharmaceutical industry, included HCS
as a strategic part of the continually evolving drug discovery
pipeline.2-4 Furthermore, HCS has already become a subject
for recurring symposia.5
Early research papers demonstrated that cellular functions
such as transcription factor activation,6 cytotoxicity,7,8 and
receptor internalization9-11 could be quantified by HCS in a
screening environment. Additional papers have described
other cellular parameters that can be screened by HCS.12-15
HCS IS AN INTEGRATED SOLUTION FOR DRUG
DISCOVERY
Two key research technologies, electronic imaging fluorescence
microscopy and fluorescence-based reagents, were
fundamental components of HCS. Imaging fluorescence microscopy
became a valuable human-interactive research tool
starting >20 years ago.16-18 Key modes of research fluorescence
microscopy that were developed over this time period
included ratio imaging,19,20 laser scanning confocal fluorescence,21
spinning disc confocal,22 deconvolution threedimensional
imaging,23 multimode microscopy,24 automated
interactive microscopy,25
standing-wave excitation fluorescence
microscopy,26 total internal reflectance microscopy,27
and multiphoton imaging.28 General research imaging software,
such as Image-Pro Plus (Media Cybernetics, Silver
Spring, MD), Metamorph (Universal Imaging, Downingtown,
PA), and Visilog (Noesis, France), has been available for smallscale
human-interactive research imaging fluorescence microscopy
for many years. These software products helped establish
image processing and analysis as a valuable tool for
cell and molecular biologists. Each of these products has an
internal programming language that enables users to create
macros or ''scripts'' to perform the analysis. Experienced users
can use these tools to design analyses to address both basic
and complex biological processes. Scripting languages give
DOI: 10.1089/154065803322302826
ª 2022 MARY ANN LIEBERT, INC. ASSAY and Drug Development Technologies 5
Assay and Drug Development Technologies

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