| Autor: |
Kengne-Ouafo JA; Department of Biochemistry, Cell and Molecular Biology, West African Centre for Cell Biology of Infectious Pathogens, University of Ghana , Accra, Ghana., Bah SY; Department of Biochemistry, Cell and Molecular Biology, West African Centre for Cell Biology of Infectious Pathogens, University of Ghana , Accra, Ghana.; Vaccine and Immunity Theme, MRC Unit The Gambia at London School of Hygiene & Tropical Medicine , Banjul, Gambia., Kemp A; Malaria Programme, Wellcome Sanger Institute, Wellcome Genome Campus , Cambridge, United Kingdom., Stewart L; Department of Infection Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine , London, United Kingdom., Amenga-Etego L; Department of Biochemistry, Cell and Molecular Biology, West African Centre for Cell Biology of Infectious Pathogens, University of Ghana , Accra, Ghana., Deitsch KW; Department of Microbiology and Immunology, Weill Medical College of Cornell University , New York City, New York, USA., Rayner JC; Malaria Programme, Wellcome Sanger Institute, Wellcome Genome Campus , Cambridge, United Kingdom.; Cambridge Institute for Medical Research, University of Cambridge , Cambridge, United Kingdom., Billker O; Malaria Programme, Wellcome Sanger Institute, Wellcome Genome Campus , Cambridge, United Kingdom.; Laboratory for Molecular Infection Medicine Sweden, Department of Molecular Biology, Umeå University , Umeå, Sweden., Binka FN; Department of Epidemiology and Biostatistics, School of Public Health, University of Health and Allied Sciences , Ho, Ghana., Sutherland CJ; Department of Infection Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine , London, United Kingdom., Awandare GA; Department of Biochemistry, Cell and Molecular Biology, West African Centre for Cell Biology of Infectious Pathogens, University of Ghana , Accra, Ghana., Urban BC; Faculty of Biological Sciences, Liverpool School of Tropical Medicine , Liverpool, United Kingdom., Dinko B; Department of Biomedical Sciences, School of Basic and Biomedical Sciences, University of Health and Allied Sciences , Ho, Ghana. |
| Abstrakt: |
Our overall understanding of the developmental biology of malaria parasites has been greatly enhanced by recent advances in transcriptomic analysis. However, most of these investigations rely on laboratory strains (LS) that were adapted into in vitro culture many years ago, and the transcriptomes of clinical isolates (CI) circulating in human populations have not been assessed. In this study, RNA-seq was used to compare the global transcriptome of mid-stage gametocytes derived from three short-term cultured CI, with gametocytes derived from the NF54 reference laboratory strain. The core transcriptome appeared to be consistent between CI- and LS-derived gametocyte preparations, but some important differences were also observed. A majority of gametocyte-specific genes (43/53) appear to have relatively higher expression in CI-derived gametocytes than in LS-derived gametocytes, but a K-means clustering analysis showed that genes involved in flagellum- and microtubule-based processes (movement/motility) were more abundant in both groups, albeit with some differences between them. In addition, gametocytes from one CI described as CI group II gametocytes (CI:GGII) showed gene expression variation in the form of reduced gametocyte-specific gene expression compared to the other two CI-derived gametocytes (CI gametocyte group I, CI:GGI), although the mixed developmental stages used in our study is a potential confounder, only partially mitigated by the inclusion of multiple replicates for each CI. Overall, our study suggests that there may be subtle differences in the gene expression profiles of mid-stage gametocytes from CI relative to the NF54 reference strain of Plasmodium falciparum . Thus, it is necessary to deploy gametocyte-producing clinical parasite isolates to fully understand the diversity of gene expression strategies that may occur during the sequestered development of parasite sexual stages. IMPORTANCE Maturing gametocytes of Plasmodium falciparum are known to sequester away from peripheral circulation into the bone marrow until they are mature. Blocking gametocyte sequestration can prevent malaria transmission from humans to mosquitoes, but most studies aim to understand gametocyte development utilizing long-term adapted laboratory lines instead of clinical isolates. This is a particular issue for our understanding of the sexual stages, which are known to decrease rapidly during adaptation to long-term culture, meaning that many LS are unable to produce transmissible gametocytes. Using RNA-seq, we investigated the global transcriptome of mid-stage gametocytes derived from three clinical isolates and a reference strain (NF54). This identified important differences in gene expression profiles between immature gametocytes of CI and the NF54 reference strain of P . falciparum , suggesting increased investment in gametocytogenesis in clinical isolates. Our transcriptomic data highlight the use of clinical isolates in studying the morphological, cellular features and molecular biology of gametocytes. |
