FAITH - Fast Assembly Inhibitor Test for HIV

Area of Science:

• Virology
• Drug Discovery
• Biochemistry

Background:

• Highly active antiretroviral therapy (HAART) for HIV-1 generates drug-resistant strains, necessitating novel therapeutic strategies.
• Inhibiting the assembly of HIV-1 Gag polyprotein into viral particles is a promising approach to block the retroviral life cycle.
• Currently, no clinically approved HIV-1 assembly inhibitors are available, partly due to a lack of efficient screening methods.

Purpose of the Study:

• To develop a novel, reliable, and simple fluorescent high-throughput screening (HTS) assay for identifying HIV-1 assembly inhibitors.
• To quantify the efficiency of retroviral particle assembly in vitro using a 96-well plate format.
• To discover compounds that inhibit the formation of both immature and mature HIV-1 particles.

Main Methods:

• Development of a fluorescent HTS assay utilizing HIV-1 Gag-derived proteins.
• Incorporation of a dual-labelled oligonucleotide that signals inhibition of particle assembly through fluorescence.
• Validation of the assay using known HIV-1 assembly inhibitors (CAI, BM2, PF74).

Main Results:

• Successfully established a new fluorescent HTS assay for quantifying HIV-1 Gag assembly efficiency in vitro.
• The assay demonstrated sensitivity in detecting inhibition of retroviral particle formation.
• The developed method was validated with three reference compounds, confirming its utility.

Conclusions:

• The novel fluorescent HTS assay provides a robust platform for identifying potential HIV-1 assembly inhibitors.
• This assay can accelerate the discovery of new anti-HIV-1 therapeutics targeting viral assembly.
• The method addresses the need for efficient screening tools to combat drug-resistant HIV-1.