Dual Host-Intracellular Parasite Transcriptome of Enucleated Cells Hosting Leishmania amazonensis: Control of Half-Life o Host Cell Transcripts by the Parasite

Author(s):Show less -

Orikaza, Cristina M. ^[1] ; Pessoa, Carina C. ^[1] ; Paladino, V, Fernanda ; Florentino, V, Pilar T. ; Barbieri, Clara L. ^[2] ; Goto, Hiro ^[3] ; Ramos-Sanchez, Eduardo Milton ^{[3, 4]} ; Silveira, Jose Franco ^[2] ; Rabinovitch, Michel ^[2] ; Mortara, Renato A. ^[2] ; Real, Fernando ^{[2, 5]}

Affiliation

^[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Sao Paulo - Brazil

^[2] Florentino, Pilar T., V, Univ Fed Sao Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Sao Paulo - Brazil

^[3] Univ Sao Paulo, Fac Med, Inst Med Trop, Lab Soroepidemiol & Imunobiol, Sao Paulo - Brazil

^[4] Univ Nacl Toribio Rodriguez de Mendoza Amazonas, Fac Ciencias Salud, Dept Salud Publ, Chachapoyas - Peru

^[5] Inst Cochin, INSERM, U1016, Dept Infect, Immunite, Inflammat, Paris - France

• Document type: Journal article
• Source: Infection and Immunity; v. 88, n. 11 NOV 2020.
• Web of Science Citations: 0

Abstract

Enucleated cells or cytoplasts (cells whose nucleus is removed in vitro) represent an unexplored biological model for intracellular infection studies due to the abrupt interruption of nuclear processing and new RNA synthesis by the host cell in response to pathogen entry. Using enucleated fibroblasts hosting the protozoan parasite Leishmania amazonensis, we demonstrate that parasite multiplication and biogenesis of large parasitophorous vacuoles in which parasites multiply are independent of the host cell nucleus. Dual RNA sequencing of both host cytoplast and intracellular parasite transcripts identified host transcripts that are more preserved or degraded upon interaction with parasites and also parasite genes that are differentially expressed when hosted by nucleated or enucleated cells. Cytoplasts are suitable host cells, which persist in culture for more than 72 h and display functional enrichment of transcripts related to mitochondrial functions and mRNA translation. Crosstalk between nucleated host de novo gene expression in response to intracellular parasitism and the parasite gene expression to counteract or benefit from these host responses induces a parasite transcriptional profile favoring parasite multiplication and aerobic respiration, and a host-parasite transcriptional landscape enriched in host cell metabolic functions related to NAD, fatty acid, and glycolytic metabolism. Conversely, interruption of host nucleus-parasite cross talk by infection of enucleated cells generates a host-parasite transcriptional landscape in which cytoplast transcripts are enriched in phagolysosome-related pathway, prosurvival, and SerpinB-mediated immunomodulation. In addition, predictive in since analyses indicated that parasite transcript products secreted within cytoplasts interact with host transcript products conserving the host V-ATPase proton translocation function and glutamine/proline metabolism. The collective evidence indicates parasite-mediated control of host cell transcripts half-life that is beneficial to parasite intracellular multiplication and escape from host immune responses. These findings will contribute to improved drug targeting and serve as database for L. arnazonensis-host cell interactions. (AU)

• FAPESP's process: 14/25296-2 - Identification of the minimum requirements for the survival of Leishmania in host cells: RNA-seq of enucleated L929 fibroblasts infected with Leishmania amazonensis
• Grantee: Renato Arruda Mortara
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/15000-4 - Trypanosoma cruzi: intra and interspecific genomic variability and mechanisms of cell invasion/egress
• Grantee: Renato Arruda Mortara
• Support Opportunities: Research Projects - Thematic Grants