Sarah D'Alessandro1, Grazia Camarda2, Yolanda Corbett1, Giulia Siciliano2, Silvia Parapini1, Luca Cevenini3, Elisa Michelini4, Aldo Roda4, Didier Leroy5, Donatella Taramelli1, Pietro Alano6. 1. Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Milano, Italy. 2. Dipartimento di Malattie Infettive, Parassitarie, Immunomediate, Istituto Superiore di Sanità, Rome, Italy. 3. Dipartimento di Chimica 'G. Ciamician', Università di Bologna, Bologna, Italy. 4. Dipartimento di Chimica 'G. Ciamician', Università di Bologna, Bologna, Italy Istituto Nazionale Biostrutture e Biosistemi (INBB), Roma, Italy. 5. Medicines for Malaria Venture, Geneva, Switzerland. 6. Dipartimento di Malattie Infettive, Parassitarie, Immunomediate, Istituto Superiore di Sanità, Rome, Italy alano@iss.it.
Abstract
OBJECTIVES: As most available antimalarial drugs are ineffective against the Plasmodium falciparum transmission stages, new drugs against the parasite's gametocytes are urgently needed to combat malaria globally. The unique biology of gametocytes requires assays that need to be specific, to faithfully monitor anti-gametocyte activity, and to be easy to perform, cheap and scalable to high-throughput screening (HTS). METHODS: We developed an HTS cell-based assay with P. falciparum gametocytes specifically expressing a potent luciferase. To confirm HTS hit activity for several parasite genotypes, the luciferase assay and the gametocyte lactate dehydrogenase (LDH) assay, usable on any parasite isolate, were compared by screening antimalarial drugs and determining IC50 values of anti-gametocyte hits from the 'Malaria Box' against early- and late-stage gametocytes. RESULTS: Comparison of the two assays, conducted on the early and on late gametocyte stages, revealed an excellent correlation (R(2) > 0.9) for the IC50 values obtained by the respective readouts. Differences in susceptibility to drugs and compounds between the two parasite developmental stages were consistently measured in both assays. CONCLUSIONS: This work indicates that the luciferase and gametocyte LDH assays are interchangeable and that their specific advantages can be exploited to design an HTS pipeline leading to new transmission-blocking compounds. Results from these assays consistently defined a gametocyte chemical susceptibility profile, relevant to the planning of future drug discovery strategies.
OBJECTIVES: As most available antimalarial drugs are ineffective against the Plasmodium falciparum transmission stages, new drugs against the parasite's gametocytes are urgently needed to combat malaria globally. The unique biology of gametocytes requires assays that need to be specific, to faithfully monitor anti-gametocyte activity, and to be easy to perform, cheap and scalable to high-throughput screening (HTS). METHODS: We developed an HTS cell-based assay with P. falciparum gametocytes specifically expressing a potent luciferase. To confirm HTS hit activity for several parasite genotypes, the luciferase assay and the gametocyte lactate dehydrogenase (LDH) assay, usable on any parasite isolate, were compared by screening antimalarial drugs and determining IC50 values of anti-gametocyte hits from the 'Malaria Box' against early- and late-stage gametocytes. RESULTS: Comparison of the two assays, conducted on the early and on late gametocyte stages, revealed an excellent correlation (R(2) > 0.9) for the IC50 values obtained by the respective readouts. Differences in susceptibility to drugs and compounds between the two parasite developmental stages were consistently measured in both assays. CONCLUSIONS: This work indicates that the luciferase and gametocyte LDH assays are interchangeable and that their specific advantages can be exploited to design an HTS pipeline leading to new transmission-blocking compounds. Results from these assays consistently defined a gametocyte chemical susceptibility profile, relevant to the planning of future drug discovery strategies.
Authors: Giulia Siciliano; T R Santha Kumar; Roberta Bona; Grazia Camarda; Maria Maddalena Calabretta; Luca Cevenini; Elisabeth Davioud-Charvet; Katja Becker; Andrea Cara; David A Fidock; Pietro Alano Journal: Mol Microbiol Date: 2017-02-21 Impact factor: 3.501
Authors: Dina Coertzen; Janette Reader; Mariëtte van der Watt; Sindisiwe H Nondaba; Liezl Gibhard; Lubbe Wiesner; Peter Smith; Sarah D'Alessandro; Donatella Taramelli; Ho Ning Wong; Jan L du Preez; Ronald Wai Keung Wu; Lyn-Marie Birkholtz; Richard K Haynes Journal: Antimicrob Agents Chemother Date: 2018-07-27 Impact factor: 5.191
Authors: Juliana Calit; Irina Dobrescu; Xiomara A Gaitán; Miriam H Borges; Marisé S Ramos; Richard T Eastman; Daniel Y Bargieri Journal: Antimicrob Agents Chemother Date: 2018-10-24 Impact factor: 5.191
Authors: Wesley C Van Voorhis; John H Adams; Roberto Adelfio; Vida Ahyong; Myles H Akabas; Pietro Alano; Aintzane Alday; Yesmalie Alemán Resto; Aishah Alsibaee; Ainhoa Alzualde; Katherine T Andrews; Simon V Avery; Vicky M Avery; Lawrence Ayong; Mark Baker; Stephen Baker; Choukri Ben Mamoun; Sangeeta Bhatia; Quentin Bickle; Lotfi Bounaadja; Tana Bowling; Jürgen Bosch; Lauren E Boucher; Fabrice F Boyom; Jose Brea; Marian Brennan; Audrey Burton; Conor R Caffrey; Grazia Camarda; Manuela Carrasquilla; Dee Carter; Maria Belen Cassera; Ken Chih-Chien Cheng; Worathad Chindaudomsate; Anthony Chubb; Beatrice L Colon; Daisy D Colón-López; Yolanda Corbett; Gregory J Crowther; Noemi Cowan; Sarah D'Alessandro; Na Le Dang; Michael Delves; Joseph L DeRisi; Alan Y Du; Sandra Duffy; Shimaa Abd El-Salam El-Sayed; Michael T Ferdig; José A Fernández Robledo; David A Fidock; Isabelle Florent; Patrick V T Fokou; Ani Galstian; Francisco Javier Gamo; Suzanne Gokool; Ben Gold; Todd Golub; Gregory M Goldgof; Rajarshi Guha; W Armand Guiguemde; Nil Gural; R Kiplin Guy; Michael A E Hansen; Kirsten K Hanson; Andrew Hemphill; Rob Hooft van Huijsduijnen; Takaaki Horii; Paul Horrocks; Tyler B Hughes; Christopher Huston; Ikuo Igarashi; Katrin Ingram-Sieber; Maurice A Itoe; Ajit Jadhav; Amornrat Naranuntarat Jensen; Laran T Jensen; Rays H Y Jiang; Annette Kaiser; Jennifer Keiser; Thomas Ketas; Sebastien Kicka; Sunyoung Kim; Kiaran Kirk; Vidya P Kumar; Dennis E Kyle; Maria Jose Lafuente; Scott Landfear; Nathan Lee; Sukjun Lee; Adele M Lehane; Fengwu Li; David Little; Liqiong Liu; Manuel Llinás; Maria I Loza; Aristea Lubar; Leonardo Lucantoni; Isabelle Lucet; Louis Maes; Dalu Mancama; Nuha R Mansour; Sandra March; Sheena McGowan; Iset Medina Vera; Stephan Meister; Luke Mercer; Jordi Mestres; Alvine N Mfopa; Raj N Misra; Seunghyun Moon; John P Moore; Francielly Morais Rodrigues da Costa; Joachim Müller; Arantza Muriana; Stephen Nakazawa Hewitt; Bakela Nare; Carl Nathan; Nathalie Narraidoo; Sujeevi Nawaratna; Kayode K Ojo; Diana Ortiz; Gordana Panic; George Papadatos; Silvia Parapini; Kailash Patra; Ngoc Pham; Sarah Prats; David M Plouffe; Sally-Ann Poulsen; Anupam Pradhan; Celia Quevedo; Ronald J Quinn; Christopher A Rice; Mohamed Abdo Rizk; Andrea Ruecker; Robert St Onge; Rafaela Salgado Ferreira; Jasmeet Samra; Natalie G Robinett; Ulrich Schlecht; Marjorie Schmitt; Filipe Silva Villela; Francesco Silvestrini; Robert Sinden; Dennis A Smith; Thierry Soldati; Andreas Spitzmüller; Serge Maximilian Stamm; David J Sullivan; William Sullivan; Sundari Suresh; Brian M Suzuki; Yo Suzuki; S Joshua Swamidass; Donatella Taramelli; Lauve R Y Tchokouaha; Anjo Theron; David Thomas; Kathryn F Tonissen; Simon Townson; Abhai K Tripathi; Valentin Trofimov; Kenneth O Udenze; Imran Ullah; Cindy Vallieres; Edgar Vigil; Joseph M Vinetz; Phat Voong Vinh; Hoan Vu; Nao-Aki Watanabe; Kate Weatherby; Pamela M White; Andrew F Wilks; Elizabeth A Winzeler; Edward Wojcik; Melanie Wree; Wesley Wu; Naoaki Yokoyama; Paul H A Zollo; Nada Abla; Benjamin Blasco; Jeremy Burrows; Benoît Laleu; Didier Leroy; Thomas Spangenberg; Timothy Wells; Paul A Willis Journal: PLoS Pathog Date: 2016-07-28 Impact factor: 6.823