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A Cell-Based Assay to Assess Hemichannel Function.

Srinivasan Krishnan1, Mariana C Fiori1, Luis G Cuello1

  • 1Department of Cell Physiology and Molecular Biophysics, and Center for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, TX.

The Yale Journal of Biology and Medicine
|March 31, 2017
PubMed
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Researchers developed a novel assay to screen for drugs targeting connexin hemichannels, crucial in diseases like stroke and deafness. This simple, scalable method uses genetically modified bacteria to identify potential hemichannel inhibitors.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Connexin hemichannels are implicated in various pathologies, including deafness, stroke, and cardiac infarct, making them a significant therapeutic target.
  • Current limitations in developing hemichannel-targeting drugs stem from a lack of selective inhibitors and efficient screening assays.
  • The translational application of potential therapies is hindered by the non-selectivity and isoform non-specificity of existing inhibitors.

Purpose of the Study:

  • To develop a novel, high-throughput screening assay for identifying drugs that modulate connexin hemichannel activity.
  • To overcome the limitations of existing assays and facilitate the discovery of selective hemichannel inhibitors.

Main Methods:

  • Genetically engineered a bacterial strain deficient in potassium (K+) uptake.
Keywords:
aminoglycosidecerebrovascular accidentconnexingap junctionhigh-throughput screeninginfarction channel

Related Experiment Videos

  • Introduced functional human connexins into the modified bacterial strain.
  • Utilized cell growth in low-K+ medium as a readout for functional hemichannel activity and K+ uptake.

    • Main Results:

    • Demonstrated that the engineered bacteria exhibit growth in low-K+ medium only when functional connexin hemichannels are expressed.
    • Established a simple, robust, and scalable cell-growth-based assay for assessing hemichannel function.
    • Validated the assay's potential for high-throughput screening of potential hemichannel-active drugs.

    Conclusions:

    • The developed bacterial assay provides a powerful new tool for discovering and characterizing connexin hemichannel modulators.
    • This assay facilitates the identification of selective inhibitors, paving the way for new therapeutic strategies for hemichannel-related disorders.
    • The assay's scalability and simplicity offer a significant advancement in drug discovery for conditions linked to connexin hemichannel dysfunction.