University of Birmingham - Whole cell protein synthesis inhibition assay for high-throughput drug discovery

Start : September 2016 | Status : Active

The scientist:  Dr Gurcha is a Post-Doctoral Researcher in the Institute of Microbiology and Infection at the University of Birmingham (BHAM), UK. After graduating from the University of Newcastle with a PhD in Microbiology, he continued his academic training under the supervision of Prof Gurdyal S. Besra aimed at understanding the biochemistry and molecular genetics of cell wall assembly in Mycobacterium tuberculosis, the causative bacterial agent of tuberculosis infections. His current research focuses on developing protein synthesis inhibition assays to identify drugs against M. tuberculosis. In addition, she is also interested in hydrophobic cell surface properties of various Mycobacterium species as it may prove useful in determining the role of cell envelope to pathogenesis and treatment of mycobacterial infections.

The sponsor: University of Birmingham

Foundation funding: The Foundation is providing £102,849 in support.

GSK’s contribution: GSK is providing expertise in High Throughput Assays (HTA) and access to Biosafety Level 3 facilities and GSK’s collection of compounds.

Project Description: Our aim is to conduct a phenotypic high-throughput screen of the “TB box”, a set of known M. tuberculosis inhibitors from the GSK collection. BHAM have developed a novel whole cell reporter assay for protein synthesis inhibition in mycobacteria, such as M. bovis BCG and M. tuberculosis. This assay utilises anhydrotetracycline-inducible mCherry expression that, in the presence of a dose response ribosomal inhibitors, produces a dose-dependent reduction in fluorescent output, which can be monitored by an automatic fluorimetric plate reader. This assay will provide an insight into the mode of action of hit compounds, which can be further validated biochemically and by the generation of spontaneous resistant mutants and whole genome sequencing. This work will be used to compliment a biochemical ribosomal HTS being conducted by Prof. Sacchettini´s group, wherein phenotypic hits will be validated for target engagement in the biochemical assay at Texas A&M University.