A high-throughput approach to in vitro anti-leishmanial drug screening-rapidly short-listing biologically effective drugs for in vivo trials

• Forschungsstelle: COST
• Projektnummer: C14.0070
• Projekttitel Englisch: A high-throughput approach to in vitro anti-leishmanial drug screening-rapidly short-listing biologically effective drugs for in vivo trials

Schlüsselwörter (Englisch)

Endoparasites; drug resistance; drug screen; Leishmania; macrophage

Forschungsprogramme (Englisch)

COST-Action CM1307 - Targeted chemotherapy towards diseases caused by endoparasites

Kurzbeschreibung (Englisch)

Leishmaniases have persisted for centuries as life threatening and disfiguring parasitic diseases. Albeit disfiguring and often disabling, cutaneous leishmaniasis is generally self-healing or responsive to 1st line therapies. A growing proportion, however are progressing to visceral or metastatic infections that are difficult to treat and more frequently plagued by symptomatic relapse. There is a clear need for novel strategies to promote and prolong the activity of anti-leishmanials. Current high-throughput drug screening efforts often focus on direct molecular interactions between putative drug targets as a preliminary indication of their parasitotoxicity, while overlooking the interactions of the host's metabolic and immune response, which may reduce drug efficacy. This proposal aims at implementing a biologically relevant process that will better gauge efficacy in the complex environment of an active infection by focusing on the effect of drugs on parasite metabolism. This process is made possible using photo-converting fluorescent transfected parasites. The quantification of the fluorescence recovery after photo-conversion subsequent to infection permits to mark parasite metabolic activity. The measurement of drug impact on pathogen metabolic activity enables the reliable evaluation of drug efficacy by a variety of techniques measurable on parasite culture as well as within an infected host.

Partner und Internationale Organisationen (Englisch)

BE; HR; CZ; EE; FR; DE; EL; IE; IL; IT; LT; MT; NL; PL; PT; RO: RS; SK; SI; ES; SE; TR; UK; US; IN; UY

Abstract (Englisch)

Our proposal fs aimed at implementing a high-throughput biologically relevant approach that will beffer gauge efficacy in the complex environment of an active infection by focusing on the effect of drugs on parasite metabolism in culture as weil as in the infected host using photo-convertinle fluorescent transfected parasites to better understand treatment failure and foresee the potential of symptomatic relapses. During this first year, we optimized a high content drug screening protocol for Leishmania RNA virus containing Leishmania guyanensis parasites within the primary murine macrophages and a mouse model of leishmaniasis fot in vivo drug testing. We performed two independent drug screening using FDA approved Prestwick Drug Library at 10 mM concentration. We identified 41 compounds out of 1280 displaying overfifty percent parasite killing activity. Currently. we are sorting compounds that will be used in hit verification experiments using photo-convertible fluorescent transcfeted parasites. We established a drug testing protocol using the only oral FDA apptoved anti leishmanial drug, We also äeveloped an in vivo drug testing protocol to fight metastatic human leishmaniasis. This approach will enable to further test efficacy and efficiency of our cornpounds. Taken together, we implemented a drug-screening pipeline, covering the in vitro and in vivo aspect ofthe leishrnaniasis.

Datenbankreferenzen (Englisch)

Swiss Database: COST-DB of the State Secretariat for Education and Research Hallwylstrasse 4 CH-3003 Berne, Switzerland Tel. [phone] Swiss Project-Number: C14.0070