Development of NanoLuc-PEST expressing Leishmania mexicana as a new drug discovery tool for axenic- and intramacrophage-based assays

PLoS Neglected Tropical Diseases
Sarah L BerryHelen P Price

Abstract

The protozoan parasite Leishmania causes leishmaniasis; a spectrum of diseases of which there are an estimated 1 million new cases each year. Current treatments are toxic, expensive, difficult to administer, and resistance to them is emerging. New therapeutics are urgently needed, however, screening the infective amastigote form of the parasite is challenging. Only certain species can be differentiated into axenic amastigotes, and compound activity against these does not always correlate with efficacy against the parasite in its intracellular niche. Methods used to assess compound efficacy on intracellular amastigotes often rely on microscopy-based assays. These are laborious, require specialist equipment and can only determine parasite burden, not parasite viability. We have addressed this clear need in the anti-leishmanial drug discovery process by producing a transgenic L. mexicana cell line that expresses the luciferase NanoLuc-PEST. We tested the sensitivity and versatility of this transgenic strain, in comparison with strains expressing NanoLuc and the red-shifted firefly luciferase. We then compared the NanoLuc-PEST luciferase to the current methods in both axenic and intramacrophage amastigotes following treatment with ...Continue Reading

References

Aug 1, 1980·International Journal of Cancer. Journal International Du Cancer·S TsuchiyaK Tada
Jul 1, 1996·Trends in Biochemical Sciences·M Rechsteiner, S W Rogers
Apr 1, 1997·Antimicrobial Agents and Chemotherapy·H L CallahanM Grogl
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Dec 30, 2003·Journal of Medicinal Chemistry·Antti MäntyläTomi Järvinen
Mar 16, 2007·Assay and Drug Development Technologies·Frank Fan, Keith V Wood
Apr 6, 2007·Molecular and Biochemical Parasitology·Gabriela BurkardIsabel Roditi
Feb 2, 2010·Nature Chemical Biology·Tilman Schneider-PoetschJun O Liu
Oct 12, 2010·PLoS Neglected Tropical Diseases·Daniel P DepledgeDeborah F Smith
Feb 26, 2011·Immunological Reviews·L David Sibley
Jul 13, 2011·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Eric H EklandDavid A Fidock
Aug 5, 2011·Parasitology·Yolanda Pérez-PertejoRosa M Reguera
Aug 24, 2011·Future Medicinal Chemistry·Robert T JacobsRobert Don
Nov 5, 2011·Assay and Drug Development Technologies·Pulin CheQianjun Li
Nov 22, 2011·The Journal of Antimicrobial Chemotherapy·Nuria AndreuBrian D Robertson
Dec 20, 2012·PloS One·Samuel O OyolaDeborah F Smith
Feb 12, 2013·Malaria Journal·Sandra HasenkampPaul Horrocks
Mar 1, 2013·Open Biology·Elizabeth BilslandStephen G Oliver
Apr 11, 2013·Antimicrobial Agents and Chemotherapy·Manu De RyckerDavid W Gray
Oct 1, 2013·Bioorganic & Medicinal Chemistry Letters·Tsutomu AkamaJacob J Plattner
Nov 28, 2013·PLoS Neglected Tropical Diseases·Alex P McLatchieMartin C Taylor
Jan 8, 2014·Antimicrobial Agents and Chemotherapy·Paola Vacchina, Miguel A Morales
Apr 11, 2014·Journal of Clinical Microbiology·Marine GinouvesGhislaine Prevot
Jul 8, 2010·ACS Medicinal Chemistry Letters·Dazhong DingJacob J Plattner
Jul 25, 2014·The Journal of Veterinary Medical Science·Keisuke SuganumaNoboru Inoue
Aug 15, 2014·Journal of Biomolecular Screening·P W Denny, P G Steel
Jun 17, 2015·BioMed Research International·Juliana Perrone Bezerra de MenezesPatrícia Sampaio Tavares Veras
Oct 6, 2015·Nature Reviews. Drug Discovery·Kei KatsunoB T Slingsby

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Citations

Sep 15, 2020·Frontiers in Pharmacology·Abdelhalim Babiker MahmoudPascal Mäser
Apr 14, 2020·Drug Design, Development and Therapy·Markos TadeleAsrat Hailu
May 13, 2021·Acta Tropica·Geovane Dias-LopesCarlos Roberto Alves
Jul 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Hippolyt L GreveThomas J Schmidt

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Methods Mentioned

BETA
flow cytometry
transgenic
PCR
Assay

Software Mentioned

MMV Pathogen Box
GraphPad Prism

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