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Development of an Image-Based HCS-Compatible Method for Endothelial Barrier Function Assessment.

Oleksii Dubrovskyi1, Erica Hasten2, Steven M Dudek3

  • 1UICentre, College of Pharmacy, University of Illinois in Chicago, Chicago, IL, USA.

SLAS Discovery : Advancing Life Sciences R & D
|July 16, 2021
PubMed
Summary
This summary is machine-generated.

Phenotypic drug discovery (PDD) offers a promising avenue for treating acute respiratory distress syndrome (ARDS). A new image cytometry assay effectively measures endothelial barrier function, aiding in the search for ARDS therapies.

Keywords:
Celigoacute respiratory distress syndrome (ARDS)cell permeabilityeXpress Permeability Test (XPerT)endothelial barrier functionimage cytometry

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Area of Science:

  • Drug discovery
  • Cell biology
  • Respiratory medicine

Background:

  • Acute respiratory distress syndrome (ARDS) is a severe condition with high mortality, exacerbated by COVID-19.
  • Current treatments for ARDS are limited, with no efficient pharmacological therapies available.
  • Increased endothelial permeability is a key factor in ARDS pathogenesis, but its regulatory mechanisms are not fully understood.

Purpose of the Study:

  • To develop a novel cell-based assay for high-throughput screening (HTS) and high-content screening (HCS) to identify potential ARDS treatments.
  • To establish an image cytometry method for measuring endothelial barrier function and cell permeability.
  • To support phenotypic drug discovery (PDD) approaches for ARDS, which are effective when molecular targets are unclear.

Main Methods:

  • Development of a plate-based image cytometry assay to assess endothelial barrier function.
  • Utilizing digital image analysis to reduce assay variability and enhance signal detection.
  • Simultaneous measurement of cell monolayer permeability and cytological characteristics.

Main Results:

  • The developed image cytometry method demonstrated reduced variability and an increased signal window compared to traditional methods.
  • The assay accurately reflected known changes in endothelial permeability in human pulmonary artery endothelial cells (HPAECs) upon stimulation with thrombin and tumor necrosis factor α.
  • The method's results correlated with established transendothelial electrical resistance (TER) measurements.

Conclusions:

  • The new image cytometry assay is a robust and adaptable tool for measuring endothelial barrier function.
  • This method facilitates rapid, simultaneous assessment of permeability and cell characteristics, suitable for HTS/HCS.
  • The assay holds significant potential for advancing phenotypic drug discovery efforts in ARDS and other conditions involving endothelial barrier dysfunction.