Heat Shock Modulates The Expression of Sirtuins and Var Genes in The Malaria Parasite Plasmodium Falciparum

Background: In the malaria parasite Plasmodium falciparum the expression of ‘var’ virulence genes is regulated through epigenetic mechanisms. Two deacetylase enzymes of the sirtuin family have been implicated in this epigenetic control in laboratory-adapted parasites. A previous study of var gene expression in parasites isolated directly from Gambian malaria patients found that high expression levels of severe-disease-associated var variants correlated with high expression of the PfSir2A sirtuin, and these expression patterns also correlated with patient phenotypes of fever and hyperlactataemia. Together, the observations suggest a mechanism through which stress phenotypes in the human host might be sensed via a parasite sirtuin, and virulence gene expression modulated accordingly. Methods: In vitro experiments were conducted using recently-laboratory-adapted Kenyan isolates of P. falciparum to follow up the correlative findings of the field study. To investigate a potential cause-and-effect relationship between host stress factors and parasite gene expression, qPCR was used to measure the expression of sirtuins and var genes after cultured parasites had been exposed to 2h or 6h of heat shock at 40°C or elevated lactate at 5mM.Results: Heat shock was shown to influence the expression of both sirtuins and var genes, whereas exposure to lactate was not. Heat shock in the trophozoite stage resulted in modest upregulation of the expression of sirtuins, particularly PfSir2B, by 2-3 fold in all strains tested. Interestingly, when heat shock was applied in ring stages PfSir2A was still upregulated but PfSir2B was downregulated. This correlated with a general upregulation of ring-stage var transcription, and particularly of severe-disease-associated upsA and upsB var genes, but there was no clear pattern in the dominant var gene(s) ultimately expressed by heat-shocked parasites. Conclusions: This study demonstrates for the first time that heat stress in recently-laboratory-adapted isolates of P. falciparum results in altered sirtuin expression – PfSir2B as well as PfSir2A – and also the upregulation of var gene expression. These may be strategies evolved by the parasite to survive heat stress when a human host experiences malarial fevers. By contrast, the association between hyperlactataemia and sirtuin/var gene expression that was previously observed in vivo appears to be coincidental rather than causative.