Biomedical Engineering Reference
In-Depth Information
FIGURE 15.1
Phospho flow is an enabling platform in drug discovery. The phospho flow
method is unique in that it can be used throughout the drug discovery process. At the target
identification stage, phospho flow can be used to generate signaling profiles to characterize
disease mechanism directly in patient samples or from animal models. For high-throughput
screening, the fluorescent cell barcoding (FCB) method and the multiparameter capabilities of
phospho flow provide novel cell-based assays to measure drug effects. At the preclinical and
clinical trial stages, the platform enables measurement of drug efficacy directly in whole blood
samples frompatients. Finally, for treatment and diagnosis of patients, phospho flowcan stratify
patients based on signaling profiles for more accurate molecular/cellular classification of
individual patient's disease states and prognosis.
flow is that the same platform can be used throughout drug discovery, such that the
same assay that was used to initially identify a lead compound can be used in
preclinical testing as well as monitoring patient response to therapy. This reduces the
need to later develop secondary assays to monitor drug action or to determine efficacy
in vivo, greatly streamlining the discovery process. In this chapter, we will explore all
these areas and the unique capabilities of the flow cytometer in advancing our
understanding of signaling networks at the single-cell level (Figure 15.1). In addition,
wewill discuss a novel hardware platform for flow cytometric analysis that is based on
mass spectrometry analysis of elemental mass tags rather than fluorescence. This
platformmay push flow cytometry into next-generation diagnostics and applications,
enabling measurement of 30-50 parameters per cell.
The chapter will be divided into sections that are summarized below.
Understanding Disease Mechanism and Target Identification Perhaps the most
important aspects of effective drug discovery are determining the molecular
causes of the disease of interest and finding appropriate drug targets. Phospho
flow enables researchers to measure the complex changes that occur in cellular
signaling networks directly from primary tissues, such as splenocytes from
murine models of disease, or directly from patient blood samples. To char-
acterize the disease, we probe the network by adding exogenous stimuli to the
cells, forcing them to respond. In this way, changes such as negative regulation
or mutations in signaling proteins can be observed and understood. These
“signaling profiles” provide a more complete view of disease pathogenesis than
does evaluation of the constitutive state of the network and a rich source of
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