Latent transcriptional variations of individual Plasmodium falciparum uncovered by single-cell RNA-seq and fluorescence imaging.

Autor: Walzer KA; Department of Molecular Genetics and Microbiology, Duke University, Durham, North Carolina, United States of America.; Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, United States of America., Fradin H; Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, United States of America., Emerson LY; Department of Molecular Genetics and Microbiology, Duke University, Durham, North Carolina, United States of America.; Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, United States of America., Corcoran DL; Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, United States of America., Chi JT; Department of Molecular Genetics and Microbiology, Duke University, Durham, North Carolina, United States of America.; Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, United States of America.
Jazyk: angličtina
Zdroj: PLoS genetics [PLoS Genet] 2019 Dec 19; Vol. 15 (12), pp. e1008506. Date of Electronic Publication: 2019 Dec 19 (Print Publication: 2019).
DOI: 10.1371/journal.pgen.1008506
Abstrakt: Malaria parasites follow a complex life cycle that consists of multiple stages that span from the human host to the mosquito vector. Among the species causing malaria, Plasmodium falciparum is the most lethal, with clinical symptoms manifesting during the intraerythrocytic developmental cycle (IDC). During the IDC, P. falciparum progresses through a synchronous and continuous cascade of transcriptional programming previously established using population analyses. While individual parasites are known to exhibit transcriptional variations to evade the host immune system or commit to a sexual fate, such rare expression heterogeneity is largely undetectable on a population level. Therefore, we combined single-cell RNA-sequencing (scRNA-seq) on a microfluidic platform and fluorescence imaging to delineate the transcriptional variations among individual parasites during late asexual and sexual stages. The comparison between asexual and sexual parasites uncovered a set of previously undefined sex-specific genes. Asexual parasites were segregated into three distinct clusters based on the differential expression of genes encoding SERAs, rhoptry proteins, and EXP2 plus transporters. Multiple pseudotime analyses revealed that these stage-specific transitions are distinct. RNA fluorescent in situ hybridization of cluster-specific genes validated distinct stage-specific expression and transitions during the IDC and defined the highly variable transcriptional pattern of EXP2. Additionally, these analyses indicated huge variations in the stage-specific transcript levels among parasites. Overall, scRNA-seq and RNA-FISH of P. falciparum revealed distinct stage transitions and unexpected degrees of heterogeneity with potential impact on transcriptional regulation during the IDC and adaptive responses to the host.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje