title: Repressing PU.1 by miR-29a∗ in NK cells of HCV patients, diminishes its cytolytic effect on HCV infected cell models.
abstract: OBJECTIVE
  Natural killer cells are immune safeguards against HCV infection. PU.1 is a pivotal transcription factor in the development of NK cells. This study aimed at studying the regulatory effect of miRNAs on both development and function of NK cells isolated from HCV patients.
  
  
  METHODS
  NK cells were isolated from 17 chronic HCV patients and 12 healthy controls; after which miRNA and mRNA were quantified using qRT-PCR. Manipulating miRNA expression using mimics and antagomirs, was performed followed by investigating downstream targets as well as viral abundance.
  
  
  RESULTS
  PU.1 expression levels were upregulated in NK cells of HCV patients. In silico analysis revealed PU.1 to be a potential downstream target of miR-29a(∗), where miR-29a(∗) overexpression in NK cells caused a significant downregulation in PU.1 mRNA. Forcing miR-29a(∗) caused a downregulation of the cytotoxicity determinant NK activating receptor (NKG2D) via upregulation of miR-155. Moreover, perforin-1 mRNA was found to be downregulated upon forcing the expression of miR-29a(∗) in NK cells of HCV patients. This decrease in NK cytolytic function was accompanied by an 80% viral load increase in cocultured HCVcc cell models.
  
  
  CONCLUSIONS
  This study showed that HCV infection might abrogate NK cytotoxic potential through altering PU.1, NKG2D receptor and perforin molecules.
authors.affiliation: The Molecular Pathology Research Group, Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Main Entrance Al Tagamoa Al Khames, 11835 Cairo, Egypt.
authors.forename: Noha Mousaad
authors.initials: NM
authors.lastname: Elemam
authors.affiliation: The Molecular Pathology Research Group, Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Main Entrance Al Tagamoa Al Khames, 11835 Cairo, Egypt.
authors.forename: Radwa Yehia
authors.initials: RY
authors.lastname: Mekky
authors.affiliation: The Molecular Pathology Research Group, Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Main Entrance Al Tagamoa Al Khames, 11835 Cairo, Egypt.
authors.forename: Nada Magdy
authors.initials: NM
authors.lastname: El-Ekiaby
authors.affiliation: The Molecular Pathology Research Group, Department of Pharmaceutical Biology, German University in Cairo, New Cairo City, Main Entrance Al Tagamoa Al Khames, 11835 Cairo, Egypt.
authors.forename: Shereen Ahmed
authors.initials: SA
authors.lastname: El Sobky
authors.affiliation: Department of Tropical Medicine and Hepatology, Cairo University, Kasr El-Aini St, Cairo, Egypt.
authors.forename: Mohammad Ahmad Mohey
authors.initials: MA
authors.lastname: El Din
authors.affiliation: Department of Tropical Medicine and Hepatology, Cairo University, Kasr El-Aini St, Cairo, Egypt.
authors.forename: Gamal
authors.initials: G
authors.lastname: Esmat
authors.affiliation: The Molecular Pathology Research Group, Department of Pharmacology and Toxicology, German University in Cairo, New Cairo City, Main Entrance Al Tagamoa Al Khames, 11835 Cairo, Egypt. Electronic address: ahmed.abdel-aziz@guc.edu.eg.
authors.forename: Ahmed Ihab
authors.initials: AI
authors.lastname: Abdelaziz
mesh_terms: D020413:3' Untranslated Regions; D016022:Case-Control Studies; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016174:Hepacivirus / Q000276:immunology*; D006526:Hepatitis C / Q000235:genetics* / Q000276:immunology*; D019698:Hepatitis C, Chronic / Q000235:genetics / Q000276:immunology; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics*; D000099232:Proto-Oncogene Protein Spi-1
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs; D055655:NK Cell Lectin-Like Receptor Subfamily K; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1; C527085:KLRK1 protein, human; C524942:MIRN155 microRNA, human; C532045:MIRN29a microRNA, human
keywords: HCV; Natural killer cells; PU.1; miRNAs