title: Laser capture microdissection enables transcriptomic analysis of dividing and quiescent liver stages of Plasmodium relapsing species.
abstract: Dormant liver stage forms (hypnozoites) of the malaria parasite Plasmodium vivax present major hurdles to control and eradicate infection. Despite major research efforts, the molecular composition of hypnozoites remains ill defined. Here, we applied a combination of state-of-the-art technologies to generate the first transcriptome of hypnozoites. We developed a robust laser dissection microscopy protocol to isolate individual Plasmodium cynomolgi hypnozoites and schizonts from infected monkey hepatocytes and optimized RNA-seq analysis to obtain the first transcriptomes of these stages. Comparative transcriptomic analysis identified 120 transcripts as being differentially expressed in the hypnozoite stage relative to the dividing liver schizont, with 69 and 51 mRNAs being up- or down-regulated, respectively, in the hypnozoites. This lead to the identification of potential markers of commitment to and maintenance of the dormant state of the hypnozoite including three transcriptional regulators of the ApiAP2 family, one of which is unique to P. cynomolgi and P. vivax, and the global translational repressor, eIF2a kinase eIK2, all of which are upregulated in the hypnozoite. Together, this work not only provides a primary experimentally-derived list of molecular markers of hypnozoites but also identifies transcriptional and posttranscriptional regulation of gene expression as potentially being key to establishing and maintaining quiescence.
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.
authors.forename: Roger
authors.initials: R
authors.lastname: Cubi
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite-Institut Pasteur, Paris, France.|CNRS ERL 9195, Paris, France.|INSERM U1201, Paris, France.
authors.forename: Shruthi S
authors.initials: SS
authors.lastname: Vembar
authors.affiliation: Centre de Bioinformatique, Biostatistique et Biologie Intégrative (C3BI, USR 3756 Institut Pasteur et CNRS), Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Biton
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.
authors.forename: Jean-Francois
authors.initials: JF
authors.lastname: Franetich
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.
authors.forename: Mallaury
authors.initials: M
authors.lastname: Bordessoulles
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.|Department of Dermatology, Eberhard Karls University, Tübingen, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Sossau
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.
authors.forename: Gigliola
authors.initials: G
authors.lastname: Zanghi
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.
authors.forename: Henriette
authors.initials: H
authors.lastname: Bosson-Vanga
authors.affiliation: Primacen, Normandie Université, Mont-Saint-Aignan, France.
authors.forename: Magalie
authors.initials: M
authors.lastname: Benard
authors.affiliation: AP-HP, Hôpital St. Antoine, Service de Parasitologie-Mycologie, 75012, Paris, France.
authors.forename: Alicia
authors.initials: A
authors.lastname: Moreno
authors.affiliation: Immunology of Viral Infections and Autoimmune Diseases, CEA-Université Paris Sud 1-INSERM U1184, Fontenay-aux-Roses, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Dereuddre-Bosquet
authors.affiliation: Immunology of Viral Infections and Autoimmune Diseases, CEA-Université Paris Sud 1-INSERM U1184, Fontenay-aux-Roses, France.
authors.forename: Roger
authors.initials: R
authors.lastname: Le Grand
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite-Institut Pasteur, Paris, France.|CNRS ERL 9195, Paris, France.|INSERM U1201, Paris, France.
authors.forename: Artur
authors.identifier: 0000-0003-2411-3328
authors.initials: A
authors.lastname: Scherf
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, CNRS ERL8255, INSERM U1135, Sorbonne Universités, UPMC Univ Paris 06, Paris, France.|AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service Parasitologie-Mycologie, Paris, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Mazier
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling; D000882:Haplorhini; D022781:Hepatocytes / Q000469:parasitology; D054884:Host-Pathogen Interactions; D061346:Laser Capture Microdissection; D008099:Liver / Q000469:parasitology*; D016789:Plasmodium cynomolgi / Q000502:physiology*
keywords: Plasmodium; hypnozoite; liver stage; quiescence