Global Proteomic Profiling of Cerebral Malaria-Infected Erythrocytes Reveals a specific PfEMP1 Expression Pattern

Cerebral malaria (CM) is the most lethal complication of malaria, causing severe impairment of brain function. Although the precise mechanisms leading to CM remain unknown, the adhesion of infected erythrocytes to the cerebral endothelium, mediated by PfEMP1 proteins, is considered a key factor. These proteins, encoded by var genes, are divided into several groups and contain specific domain cassettes (DC) already known to be associated with CM. In this study, we compared the human and parasite proteomes of Plasmodium falciparum infected erythrocytes from nineteen samples from the NeuroCM cohort, consisting of 10 children with uncomplicated malaria (UM) and 9 with CM from Benin. We used data-independent acquisition mass spectrometry (DIA-MS) with Trapped Ion Mobility Spectrometry (TIMS) and Parallel Accumulation Serial Fragmentation (PASEF) coupled to recent PfEMP1 databases to analyze our samples. Our results identified over 7,000 proteins, including both host and parasite proteins. Analysis of the host proteome revealed an increased abundance of reticulocyte-specific proteins in CM cases, suggesting a possible parasite preference for invading erythroid precursors. On the parasite side, an increase in membrane proteins (PHIST, RIFIN, STEVOR) was observed in CM cases. Moreover, PfEMP1 proteins containing DC8 and DC13 were significantly more abundant in CM patients. These results provide protein-level confirmation of the association between DC8/DC13 of PfEMP1 and CM, supporting the hypothesis of an enhanced parasite adhesion mechanism in CM. Our study highlights the importance of proteomic analysis in understanding the pathogenesis of CM and may contribute to the development of novel therapeutic strategies.