DEVELOPMENT OF A MULTIMODAL SCREENING APPROACH TOWARDS BORRELIACIDAL COMPOUNDS WITH NOVEL MECHANISM OF ACTION

Abstract

This work addresses the unmet need for effective therapies targeting Lyme disease by developing innovative platforms and methods. The research focuses on identifying small molecules capable of killing Borrelia burgdorferi, the bacterium responsible for Lyme disease, through parallel target-based and phenotypic screening strategies. Central to this effort is the investigation of the BbHtrA protein, a chaperone-protease essential for bacterial virulence, as a promising therapeutic target. We establish workflows for the expression, purification, and biophysical characterization of BbHtrA, alongside novel assay platforms to screen for modulators of its activity. A critical contribution of this work is the development of orthogonal assays for hit validation, ensuring the robustness of identified lead compounds. We also develop and optimize Borrelia viability screening assays, including luciferase-based and orthogonal assays, to identify borreliacidal compounds with several orders of magnitude stronger potency than those in the prior art. This research represents a comprehensive effort to advance early-stage drug discovery for Lyme disease, addressing current therapeutic limitations while introducing new tools, assays, and insights to the field.