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Transcriptome analysis of Plasmodium falciparum isolates from Benin reveals specific gene expression associated with cerebral malaria

ABSTRACT: Transcriptome analysis of Plasmodium falciparum isolates from Benin reveals specific gene expression associated with cerebral malaria

PROVIDER: PRJNA776243 | ENA |

REPOSITORIES: ENA

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Transcriptome analysis of Plasmodium falciparum isolates from Benin reveals specific gene expression associated with cerebral malaria

Project description:The most severe form of Plasmodium falciparum infection, cerebral malaria (CM), is linked to the parasite’s ability to bind to specific endothelial receptors in brain capillaries. However, the host and parasitic factors causing CM are not yet fully elucidated and the transcriptome profile of P. falciparum isolated from patients with CM remains largely unknown. To complement this deficiency, we aimed to characterize the gene expression associated with CM, by sequencing the parasite’s transcriptome from 15 CM and 15 uncomplicated malaria Beninese children. We demonstrated a specific gene expression in CM, making it possible to completely distinguish the two clinical phenotypes. Numerous biological pathways including entry into host and cytoadherence were predicted as impacted. In addition, infected erythrocytes (iE) from CM children had a decreased circulation time. It may be the result of a stronger adhesion capacity of these iE, which we previously demonstrated in vitro. Consistent with this result, we were able to measure an overall increased expression of the var genes family, encoding for the major mediator of adherence to host endothelium. This phenomenon may allow iE to avoid splenic clearance and cause the increased parasitaemia measured in CM.

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Discovery of telomere repeat-binding factor in the malaria parasite Plasmodium falciparum

Project description:ChIP-seq experiments were performed for the putative telomere repeat-binding factor (PfTRF) in the malaria parasite Plasmodium falciparum strain 3D7. The gene encoding this factor (PF3D7_1209300) was endogenously tagged with either a GFP- or a 3xHA-tag and these transgenic parasite lines were used in ChIP-sequencing experiments. Sequencing of the ChIP and input libraries showed enrichment of PfTRF at all telomere-repeat containing chromosome ends (reference genome Plasmodium falciparum 3D7 from PlasmoDB version 6.1) as well as in all upsB var promoters.In addition,PfTRF was enriched at seven additional, intra-chromosomal sites and called in the PfTRF-HA ChIP-seq only. Plasmodium falciparum 3D7 parasites were generated with -GFP or -3xHA C-terminal tagged TRF (PF3D7_1209300). Nuclei were isolated from formaldehyde cross-linked schizont-stage transgenic parasites and used to prepare chromatin. Chromatin immunoprecipitations were performed using mouse anti-GFP (Roche Diagnostics, #11814460001) or rat anti-HA 3F10 (Roche Diagnostics, #12158167001). Sequencing libraries were prepared according to a Plasmodium-optimized library preparation procedure including KAPA polymerase-mediated PCR amplification.

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Transcription profiling of genetically cerebral malaria-resistant (CM-R) and cerebral malaria-susceptible (CM-S) mouse strains

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Blood transcriptional profiles discriminates cerebral and mild malaria patient living in Senegal

Project description:PBMCs were obtain from mild malaria and cerebral malaria patient at the onset of P. falciparum infection. Patient recruitment took place in two hospital centers in Senegal Total RNAs from PBMCs of mild and cerebral malaria patients were profiled after hybridization with Agilent SurePrint G3 Human GE 8x60K Microarray to identify blood biomarkers for cerebral malaria phenotype Plasmodium falciparum malaria remains a major health problem in Africa. The mechanisms of pathogenesis leading to the severe form of the disease are not fully understood. Blood transcriptional profiles were investigated in patient with cerebral malaria, noncerebral malaria , and mild malaria by using the microarray technology. We identified a set of 447 genes that was differentially expressed between the three patient groups after a false discovery rate of 10%. Since the cerebral patients displayed a particular transcriptional pattern, we focused our analysis on the differences between cerebral malaria patients and mild malaria patients. We further found 849 genes differentially expressed after a false discovery rate of 10%. We validated our results by using the qPCR method for 5 genes. The enrichment analysis of their functional annotation indicates that genes involved in A, B, C pathways play a role in the occurrence of cerebral malaria. These results provide new insight into the potential effect of the dysregulation of gene expression and specific pathways. Host genetic variation may partly explain such alteration of gene expression. Further studies are required to investigate this in African populations.

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