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      KSHV oral shedding and plasma viremia result in significant changes in the extracellular tumorigenic miRNA expression profile in individuals infected with the malaria parasite

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          Abstract

          Kaposi's sarcoma herpesvirus (KSHV) is the etiological agent of Kaposi’s sarcoma (KS). Both KSHV and HIV infections are endemic in Uganda, where KS is among the most common cancers in HIV-infected individuals. Recent studies examined the use of small RNAs as biomarkers of disease, including microRNAs (miRNAs), with viral and tumor-derived miRNAs being detected in exosomes from individuals with KSHV-associated malignancies. In the current study, the host and viral extracellular mature miRNA expression profiles were analyzed in blood of KS-negative individuals in Uganda, comparing those with or without KSHV detectable from the oropharynx. We observed increased levels of cellular oncogenic miRNAs and decreased levels of tumor-suppressor miRNAs in plasma of infected individuals exhibiting oral KSHV shedding. These changes in host oncomiRs were exacerbated in people co-infected with HIV, and partially reversed after 2 years of anti-retroviral therapy. We also detected KSHV miRNAs in plasma of KSHV infected individuals and determined that their expression levels correlated with KSHV plasma viremia. Deep sequencing revealed an expected profile of small cellular RNAs in plasma, with miRNAs constituting the major RNA biotype. In contrast, the composition of small RNAs in exosomes was highly atypical with high levels of YRNA and low levels of miRNAs. Mass spectrometry analysis of the exosomes revealed eleven different peptides derived from the malaria parasite, Plasmodium falciparum, and small RNA sequencing confirmed widespread plasmodium co-infections in the Ugandan cohorts. Proteome analysis indicated an exosomal protein profile consistent with erythrocyte and keratinocyte origins for the plasma exosomes. A strong correlation was observed between the abundance of Plasmodium proteins and cellular markers of malaria. As Plasmodium falciparum is an endemic pathogen in Uganda, our study shows that co-infection with other pathogens, such as KSHV, can severely impact the small RNA repertoire, complicating the use of exosome miRNAs as biomarkers of disease.

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          HDL-transferred microRNA-223 regulates ICAM-1 expression in endothelial cells.

          High-density lipoproteins (HDL) have many biological functions, including reducing endothelial activation and adhesion molecule expression. We recently reported that HDL transport and deliver functional microRNAs (miRNA). Here we show that HDL suppresses expression of intercellular adhesion molecule 1 (ICAM-1) through the transfer of miR-223 to endothelial cells. After incubation of endothelial cells with HDL, mature miR-223 levels are significantly increased in endothelial cells and decreased on HDL. However, miR-223 is not transcribed in endothelial cells and is not increased in cells treated with HDL from miR-223(-/-) mice. HDL inhibit ICAM-1 protein levels, but not in cells pretreated with miR-223 inhibitors. ICAM-1 is a direct target of HDL-transferred miR-223 and this is the first example of an extracellular miRNA regulating gene expression in cells where it is not transcribed. Collectively, we demonstrate that HDL's anti-inflammatory properties are conferred, in part, through HDL-miR-223 delivery and translational repression of ICAM-1 in endothelial cells.
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            “Asymptomatic” Malaria: A Chronic and Debilitating Infection That Should Be Treated

            Roland Gosling and colleagues argue that "asymptomatic" malaria infections have significant health and societal consequences, and propose that they should be renamed "chronic" malaria infections.
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              A viral microRNA functions as an orthologue of cellular miR-155.

              All metazoan eukaryotes express microRNAs (miRNAs), roughly 22-nucleotide regulatory RNAs that can repress the expression of messenger RNAs bearing complementary sequences. Several DNA viruses also express miRNAs in infected cells, suggesting a role in viral replication and pathogenesis. Although specific viral miRNAs have been shown to autoregulate viral mRNAs or downregulate cellular mRNAs, the function of most viral miRNAs remains unknown. Here we report that the miR-K12-11 miRNA encoded by Kaposi's-sarcoma-associated herpes virus (KSHV) shows significant homology to cellular miR-155, including the entire miRNA 'seed' region. Using a range of assays, we show that expression of physiological levels of miR-K12-11 or miR-155 results in the downregulation of an extensive set of common mRNA targets, including genes with known roles in cell growth regulation. Our findings indicate that viral miR-K12-11 functions as an orthologue of cellular miR-155 and probably evolved to exploit a pre-existing gene regulatory pathway in B cells. Moreover, the known aetiological role of miR-155 in B-cell transformation suggests that miR-K12-11 may contribute to the induction of KSHV-positive B-cell tumours in infected patients.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: Methodology
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: Writing – review & editing
                Role: Data curationRole: Project administrationRole: ResourcesRole: Writing – review & editing
                Role: Formal analysisRole: MethodologyRole: Software
                Role: Formal analysisRole: MethodologyRole: Software
                Role: Formal analysisRole: MethodologyRole: Software
                Role: Formal analysisRole: Software
                Role: Funding acquisitionRole: Project administrationRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                9 February 2018
                2018
                : 13
                : 2
                : e0192659
                Affiliations
                [1 ] Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, Washington, United States of America
                [2 ] Department of Pediatrics, University of British Columbia, BC Children’s Hospital, Vancouver, British Columbia, Canada
                [3 ] Infectious Disease Research Institute, Seattle, Washington, United States of America
                [4 ] Departments of Medicine and Global Health, University of Washington, Seattle, Washington, United States of America
                [5 ] Proteomics, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America
                [6 ] Genomics, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America
                [7 ] Department of Pediatrics, University of Washington, Seattle, Washington, United States of America
                Institut de Pharmacologie Moleculaire et Cellulaire, FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                [¤a]

                Current address: Just biotherapeutics for all, Seattle, Washington, United States of America.

                [¤b]

                Current address: King County Environmental Laboratory, Dept. of Natural Resources and Parks, Seattle, Washington, United States of America

                Author information
                http://orcid.org/0000-0001-7148-5621
                Article
                PONE-D-17-42992
                10.1371/journal.pone.0192659
                5806893
                29425228
                2dbca411-69b1-4383-b500-997b57da9caa
                © 2018 Ikoma et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 7 December 2017
                : 26 January 2018
                Page count
                Figures: 14, Tables: 8, Pages: 46
                Funding
                Funded by: NIH/National Institute of Dental and Craniofacial Research
                Award ID: 1P01 DE021954
                Award Recipient :
                This work was supported by an NIH funded program, NIH/National Institute of Dental and Craniofacial Research, Interdisciplinary Research on Oral Manifestations of HIV/AIDS in Vulnerable Populations, 1P01 DE021954. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and life sciences
                Genetics
                Gene expression
                Gene regulation
                MicroRNAs
                Biology and life sciences
                Biochemistry
                Nucleic acids
                RNA
                Non-coding RNA
                MicroRNAs
                Biology and Life Sciences
                Anatomy
                Body Fluids
                Blood
                Blood Plasma
                Medicine and Health Sciences
                Anatomy
                Body Fluids
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                Physiology
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                Blood Plasma
                Medicine and Health Sciences
                Physiology
                Body Fluids
                Blood
                Blood Plasma
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Vesicles
                Exosomes
                Medicine and Health Sciences
                Parasitic Diseases
                Malaria
                Medicine and Health Sciences
                Tropical Diseases
                Malaria
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
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                Immunodeficiency Viruses
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                Pathogens
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                Organisms
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                Retroviruses
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                Kaposi's Sarcoma-Associated Herpesvirus
                Biology and Life Sciences
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                Kaposi's Sarcoma-Associated Herpesvirus
                Medicine and Health Sciences
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                Custom metadata
                The proteomic data were submitted to ProteomeXchange via the PRIDE database under accession number GSE1062338. Similarly, all the sequencing data were submitted to the GEO database under a single accession number: PXD008107.

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