Phenotypic Screening

Palmer, Michelle

doi:10.1002/9781118771723.ch10

Cited by 1 publication

(3 citation statements)

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“…This methodology provides more biologically-relevant information in a target-agnostic manner. 4 Phenotypic screening, generally considered to be a classical method of drug discovery, has recently seen resurgence in the field due to significant technological advancements for both target identification and small-molecule discovery. Contrary to most target-based approaches, phenotypic screens offer the ability to deliver novel and first-in-class therapeutics.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…8 After this clustering, the challenge remains to elucidate the targets and MoAs associated with the phenotypes, but this burden decreased with recent advancements including genome-wide CRISPR screens. [4][5][6] While the observation of morphological phenotypes has advanced phenotypic screens, it is understood that the observable endpoints resulting from compound treatment are caused by changes in the underlying metabolic pathways. 9,10 Many have explored mass spectrometry (MS)-based approaches such as metabolomics as ways to accomplish phenotypic drug discovery by monitoring metabolites and the associated activation of biological pathways corresponding to observed phenotypes.…”

Section: Introductionmentioning

confidence: 99%

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Metabolite Fingerprinting for Phenotypic Screening by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry

Joignant,

Pu,

McLoughlin

et al. 2023

Preprint

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The adoption of mass spectrometry for high-throughput screening in drug discovery has become increasingly prevalent and has enabled label-free screening against diverse targets. Cellular assays for phenotypic screening, however, are primarily conducted by microscopy as there remain many challenges associated with conducting phenotypic screens via ultra-high throughput mass spectrometry.Following a simple on-plate extraction, infrared matrix assisted laser desorption electrospray ionization (IR-MALDESI) was employed to directly sample the cell lysate at a speed of one second per sample. A549 cells were treated with compounds identified as hits in literature including a recently reported glutaminase cellular screen. Among the test compounds, there were confirmed glutaminase inhibitors, proposed nuisance compounds, and cell-active but enzyme-inactive compounds. Filtered data were further processed in R for dimensionality reduction and unsupervised clustering.Though it was observed that all compounds affected the intracellular conversion of glutamine to glutamate, there were clear metabolic differences between the biochemically active compounds and the off-target hits. Moreover, two nuisance compounds were observed to cluster separately from the confirmed glutaminase inhibitors in the observed metabolite fingerprints.This proof-of-concept work establishes a complete workflow that enables highthroughput mass spectrometry-based phenotypic screening. The methods proposed herein, at the throughput enabled by IR-MALDESI, could offer a new avenue for discovery of novel drugs.

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Section: Introductionmentioning

confidence: 99%