Whole‐Transcriptome Analysis of<i>Plasmodium falciparum</i>Field Isolates: Identification of New Pathogenicity Factors

Siau, Anthony; Touré, Fousseyni S.; Ouwe-Missi-Oukem-Boyer, Odile; Ciceron, Liliane; Mahmoudi, Nassira; Vaquero, Catherine; Froissard, Patrick; Bisvigou, Ulrick; Bisser, Sylvie; Coppée, Jean‐Yves; Bischoff, Emmanuel; David, Peter H.; Mazier, Dominique

doi:10.1086/522012

Cited by 29 publications

(37 citation statements)

References 46 publications

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“…One way to approach the analysis of parasite-specific factors in disease severity is to compare the transcriptional profiles of parasites taken directly from patients with different clinical presentations. Several such analyses have already been carried out but with conflicting conclusions (1)(2)(3)(4).…”

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confidence: 99%

Statistical estimation of cell-cycle progression and lineage commitment in Plasmodium falciparum reveals a homogeneous pattern of transcription in ex vivo culture

Lemieux

Gomez-Escobar

Feller

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

135

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We have cultured Plasmodium falciparum directly from the blood of infected individuals to examine patterns of mature-stage gene expression in patient isolates. Analysis of the transcriptome of P. falciparum is complicated by the highly periodic nature of gene expression because small variations in the stage of parasite development between samples can lead to an apparent difference in gene expression values. To address this issue, we have developed statistical likelihood-based methods to estimate cell cycle progression and commitment to asexual or sexual development lineages in our samples based on microscopy and gene expression patterns. In cases subsequently matched for temporal development, we find that transcriptional patterns in ex vivo culture display little variation across patients with diverse clinical profiles and closely resemble transcriptional profiles that occur in vitro. These statistical methods, available to the research community, assist in the design and interpretation of P. falciparum expression profiling experiments where it is difficult to separate true differential expression from cell-cycle dependent expression. We reanalyze an existing dataset of in vivo patient expression profiles and conclude that previously observed discrete variation is consistent with the commitment of a varying proportion of the parasite population to the sexual development lineage. malaria ͉ microarray

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mentioning

confidence: 99%

Statistical estimation of cell-cycle progression and lineage commitment in Plasmodium falciparum reveals a homogeneous pattern of transcription in ex vivo culture

Lemieux

Gomez-Escobar

Feller

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

135

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“…The applicability of RNA interference (RNAi), which has been extremely important in elucidating gene function in several eukaryotic organisms, is still being debated. Several groups have reported successfully using double-stranded RNA to specifically knock down target genes such as falcipains-1 and 2 (6, 7), dihydroorotate dehydrogenase (8), PfMyb1, a transcription factor (9), and several genes putatively involved in parasite infected RBC induction of endothelial cell apoptosis (10). However, whether this is mediated via RNAi per se or anti-sense mechanisms remains unclear, especially in light of bioinformatics data suggesting that key conserved components of the RNAi machinery are not encoded in the P. falciparum genome (11,12).…”

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confidence: 99%

Inhibiting Plasmodium falciparum growth and heme detoxification pathway using heme-binding DNA aptamers

Niles¹,

DeRisi

Marletta

2009

Proc. Natl. Acad. Sci. U.S.A.

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The human parasite Plasmodium falciparum enzymatically digests hemoglobin during its intra-erythrocytic developmental stages in acidic food vacuole compartments. The released heme is rapidly detoxified by polymerization into the chemically inert pigment, hemozoin. Several heme-binding anti-malarial compounds, such as chloroquine, efficiently inhibit this process, and this is believed to be the predominant mechanism by which these drugs induce parasite toxicity. In an effort to expand the biochemical tools available for exploration of this pathogen's basic biology, we chose this heme-detoxification pathway as a model system for exploring the suitability of DNA aptamers for modulating this essential parasite biochemical pathway. In this report, we demonstrate that heme-binding DNA aptamers efficiently inhibit in vitro hemozoin formation catalyzed by either a model lipid system or parasite-derived extracts just as or more potently than chloroquine. Furthermore, when parasites are grown in red cells loaded with heme-binding aptamers, their growth is significantly inhibited relative to parasites exposed to non-heme-binding DNA oligonucleotides. Both the timing of parasite-induced toxicity and the concentration of heme-binding aptamer required for inducing toxicity correlate well with the uptake of red cell cytosolic components by the parasite, and the requirement for compounds with similar in vitro hemozoin inhibitory potency to preconcentrate within the parasite before observing toxicity. Thus, these hemebinding aptamers recapitulate the in vitro hemozoin inhibition activity and induce parasite toxicity in a manner consistent with inhibition of this pathway. Altogether, these data demonstrate that aptamers can be versatile tools with applicability in functionally dissecting important P. falciparum-specific pathways both in vitro and in vivo.

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“…Optical density (OD) was read with a Stat FaxH 3200 (Fisher Bioblock Scientific, Illkirch France) at 405 nm, with a reference filter of 492 nm. The mean OD of pRBC-activated HLEC and hCMEC/D3 to RBC-activated HLEC and hCMEC/D3 and mean OD of 3D7-activated HLEC and hCMEC/D3 to RBC-activated HLEC and hCMEC/D3 was calculated, using a positivity cut-off of 3, as recommended [6, 14]. …”

Section: Methodsmentioning

confidence: 99%

“…Using cocultured human lung EC (HLEC) and pRBC field isolates from Gabon, we showed that although almost all isolates provoked cytoadherence, only a few triggered apoptosis (via cytoadherence or diffusible factors), and that these isolates tended to have been drawn from patients with neurological disorders [9]. The potential of pRBC to trigger HLEC apoptosis varies according to the isolate, and is associated to the expression of Plasmodium apoptosis-linked pathogenicity factors (PALPFs) [9, 14, 15]. Human endothelial cells in different tissues may not all express the same surface receptors [16], and this could explain some functional differences.…”

Section: Introductionmentioning

confidence: 99%

Comparison of apoptosis in human primary pulmonary endothelial cells and a brain microvascular endothelial cell line co-cultured with Plasmodium falciparum field isolates

et al. 2017

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Background Plasmodium falciparum infection can progress unpredictably to severe forms including respiratory distress and cerebral malaria. The mechanisms underlying the variable natural course of malaria remain elusive.MethodsThe cerebral microvascular endothelial cells-D3 and lung endothelial cells both from human were cultured separately and challenged with P. falciparum field isolates taken directly from malaria patients or 3D7 strain (in vitro maintained culture). The capacity of these P. falciparum isolates to induce endothelial cell apoptosis via cytoadherence or not was then assessed.ResultsOverall, 27 P. falciparum isolates were collected from patients with uncomplicated malaria (n = 25) or severe malaria (n = 2). About half the isolates (n = 17) were able to bind brain endothelial cells (12 isolates, 44%) or lung endothelial cells (17 isolates, 63%) or both (12 isolates, 44%). Sixteen (59%) of the 27 isolates were apoptogenic for brain and/or lung endothelial cells. The apoptosis stimulus could be cytoadherence, direct cell-cell contact without cytoadherence, or diffusible soluble factors. While some of the apoptogenic isolates used two stimuli (direct contact with or without cytoadherence, plus soluble factors) to induce apoptosis, others used only one. Among the 16 apoptogenic isolates, eight specifically targeted brain endothelial cells, one lung endothelial cells, and seven both.ConclusionThese results indicate that the brain microvascular cell line was more susceptible to apoptosis triggered by P. falciparum than the primary pulmonary endothelial cells and may have relevance to host-parasite interaction.

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Whole‐Transcriptome Analysis ofPlasmodium falciparumField Isolates: Identification of New Pathogenicity Factors

Abstract: This is the first attempt to identify genes involved in parasite pathogenic mechanisms against human ECs. The finding of PALPFs illuminates perspectives for novel therapeutic strategies against cerebral complications of malaria.

Cited by 29 publications

References 46 publications

Statistical estimation of cell-cycle progression and lineage commitment in Plasmodium falciparum reveals a homogeneous pattern of transcription in ex vivo culture

Statistical estimation of cell-cycle progression and lineage commitment in Plasmodium falciparum reveals a homogeneous pattern of transcription in ex vivo culture

Inhibiting Plasmodium falciparum growth and heme detoxification pathway using heme-binding DNA aptamers

Comparison of apoptosis in human primary pulmonary endothelial cells and a brain microvascular endothelial cell line co-cultured with Plasmodium falciparum field isolates

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