Justin Gullingsrud1, Neta Milman1, Tracy Saveria1, Olga Chesnokov2, Kathryn Williamson1, Anand Srivastava3, Benoit Gamain3, Patrick E Duffy4, Andrew V Oleinikov5. 1. Seattle Biomedical Research Institute, Seattle, Washington. 2. Charles E. Schmidt College of Medicine, Department of Biomedical Science, Florida Atlantic University, Boca Raton. 3. Inserm UMR 1134 Université Paris Diderot, Sorbonne Paris Cité, UMR S1134 Institut National de la Transfusion Sanguine, Paris, France. 4. Laboratory of Malaria Immunology and Vaccinology, National Institutes of Health, Bethesda, Maryland. 5. Seattle Biomedical Research Institute, Seattle, Washington Charles E. Schmidt College of Medicine, Department of Biomedical Science, Florida Atlantic University, Boca Raton.
Abstract
BACKGROUND: We developed a 2-step approach to screen molecules that prevent and/or reverse Plasmodium falciparum-infected erythrocyte (IE) binding to host receptors. IE adhesion and sequestration in vasculature causes severe malaria, and therefore antiadhesion therapy might be useful as adjunctive treatment. IE adhesion is mediated by the polymorphic family (approximately 60 members) of P. falciparum EMP1 (PfEMP1) multidomain proteins. METHODS: We constructed sets of PfEMP1 domains that bind ICAM-1, CSA, or CD36, receptors that commonly support IE binding. Combinations of domain-coated beads were assayed by Bio-Plex technology as a high-throughput molecular platform to screen antiadhesion molecules (antibodies and small molecules). Molecules identified as so-called hits in the screen (first step) then could be assayed individually for inhibition of binding of live IE to receptors (second step). RESULTS: In proof-of-principle studies, the antiadhesion activity of several antibodies was concordant in Bio-Plex and live IE assays. Using this 2-step approach, we identified several molecules in a small molecule library of 10 000 compounds that could inhibit and reverse binding of IEs to ICAM-1 and CSA receptors. CONCLUSION: This 2-step screening approach should be efficient for identification of antiadhesion drug candidates for falciparum malaria. Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US.
BACKGROUND: We developed a 2-step approach to screen molecules that prevent and/or reverse Plasmodium falciparum-infected erythrocyte (IE) binding to host receptors. IE adhesion and sequestration in vasculature causes severe malaria, and therefore antiadhesion therapy might be useful as adjunctive treatment. IE adhesion is mediated by the polymorphic family (approximately 60 members) of P. falciparum EMP1 (PfEMP1) multidomain proteins. METHODS: We constructed sets of PfEMP1 domains that bind ICAM-1, CSA, or CD36, receptors that commonly support IE binding. Combinations of domain-coated beads were assayed by Bio-Plex technology as a high-throughput molecular platform to screen antiadhesion molecules (antibodies and small molecules). Molecules identified as so-called hits in the screen (first step) then could be assayed individually for inhibition of binding of live IE to receptors (second step). RESULTS: In proof-of-principle studies, the antiadhesion activity of several antibodies was concordant in Bio-Plex and live IE assays. Using this 2-step approach, we identified several molecules in a small molecule library of 10 000 compounds that could inhibit and reverse binding of IEs to ICAM-1 and CSA receptors. CONCLUSION: This 2-step screening approach should be efficient for identification of antiadhesion drug candidates for falciparum malaria. Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US.
Entities:
Keywords:
CD36; CSA; ICAM-1; PfEMP1 proteins; Plasmodium falciparum; antiadhesion therapy; high-throughput screening; host receptors; malaria; sequestration of parasites; small molecules
Authors: Pamela A Magistrado; Daniel Minja; Justin Doritchamou; Nicaise Tuikue Ndam; Davis John; Christentze Schmiegelow; Achille Massougbodji; Madeleine Dahlbäck; Sisse B Ditlev; Vera V Pinto; Mafalda Resende; John Lusingu; Thor G Theander; Ali Salanti; Morten A Nielsen Journal: Vaccine Date: 2010-11-12 Impact factor: 3.641
Authors: Morten A Nielsen; Vera V Pinto; Mafalda Resende; Madeleine Dahlbäck; Sisse B Ditlev; Thor G Theander; Ali Salanti Journal: Infect Immun Date: 2009-03-23 Impact factor: 3.441
Authors: Joel H Janes; Christopher P Wang; Emily Levin-Edens; Inès Vigan-Womas; Micheline Guillotte; Martin Melcher; Odile Mercereau-Puijalon; Joseph D Smith Journal: PLoS Pathog Date: 2011-05-05 Impact factor: 6.823
Authors: Jun-Hong Ch'ng; Kirsten Moll; Maria Del Pilar Quintana; Sherwin Chun Leung Chan; Ellen Masters; Ernest Moles; Jianping Liu; Anders B Eriksson; Mats Wahlgren Journal: Sci Rep Date: 2016-07-11 Impact factor: 4.379