title: Effects of the vascular endothelial growth factor receptor-2 (VEGFR-2) inhibitor SU5416 on in vitro cultures of Plasmodium falciparum.
abstract: BACKGROUND
  Vascular endothelial growth factor (VEGF) is taken up by parasitized red blood cells during malaria and stimulates intra-erythrocytic growth of Plasmodium falciparum in vitro. The cause and consequence of this uptake is not understood.
  
  
  METHODS
  Plasmodium falciparum was cultured in vitro. Parasite growth and intracellular VEGF levels were assessed using flow cytometry. Intracellular VEGF was visualized by fluorescence immunocytochemistry. Phosphorylated tyrosine was measured by western blotting. In vivo assessment of intra-erythrocytic VEGF was performed in Plasmodium berghei ANKA-infected C57BL/6 mice.
  
  
  RESULTS
  VEGF accumulated intracellularly in infected red blood cells, particularly in schizonts. In vitro growth of P. falciparum was unchanged when co-cultured with the anti-VEGF antibody bevacizumab or with an anti-VEGF receptor-1 peptide. In contrast, the VEGF receptor-2 inhibitor, SU5416, dose-dependently inhibited growth. None of the treatments reduced intracellular VEGF levels. Thus, the anti-parasitic effect of SU5416 seemed independent of VEGF uptake. SU5416 reduced phosphorylated tyrosine in parasitized red blood cells. Similarly, the broad-spectrum tyrosine kinase inhibitor genistein dose-dependently inhibited P. falciparum growth and reduced tyrosine phosphorylation. Neither bevacizumab nor anti-VEGF receptor-1 peptide affected tyrosine kinase activity. Finally, in vivo uptake of VEGF in P. berghei ANKA was demonstrated, analogous to the in vitro uptake in P. falciparum, making it a possible model for the effects of VEGF signalling in vivo during malaria.
  
  
  CONCLUSIONS
  Inhibition of VEGFR-2 signalling reduces intra-erythrocytic growth of P. falciparum, likely due to tyrosine kinase inhibition. Internalisation of VEGF in P. falciparum-infected red blood cells does not rely on VEGF receptors. The function of in vivo uptake of VEGF can be studied in rodent malaria models.
authors.forename: Casper
authors.initials: C
authors.lastname: Hempel
authors.affiliation: Centre for Medical Parasitology, Department of Clinical Microbiology, Copenhagen University Hospital, Copenhagen, Denmark. nils.hoyer@gmail.com.
authors.forename: Nils
authors.initials: N
authors.lastname: Hoyer
authors.forename: Trine
authors.initials: T
authors.lastname: Staalsø
authors.forename: Jørgen A
authors.initials: JA
authors.lastname: Kurtzhals
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D003600:Cytosol / Q000737:chemistry; D004195:Disease Models, Animal; D004912:Erythrocytes / Q000737:chemistry* / Q000469:parasitology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles / Q000494:pharmacology*; D008288:Malaria / Q000188:drug therapy / Q000469:parasitology; D008810:Mice, Inbred C57BL; D010962:Plasmodium berghei / Q000254:growth & development; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000254:growth & development*; D011758:Pyrroles / Q000494:pharmacology*; D014443:Tyrosine / Q000032:analysis; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis / Q000037:antagonists & inhibitors*
chemical_list: D000962:Antimalarials; D007211:Indoles; D011758:Pyrroles; D014443:Tyrosine; C116890:Semaxinib; D040301:Vascular Endothelial Growth Factor Receptor-2