title: miR-29b and miR-198 overexpression in CD8+ T cells of renal cell carcinoma patients down-modulates JAK3 and MCL-1 leading to immune dysfunction.
abstract: BACKGROUND
  Mammalian microRNAs (miR) regulate the expression of genes relevant for the development of adaptive and innate immunity against cancer. Since T cell dysfunction has previously been reported in patients with renal cell carcinoma (RCC; clear cell type), we aimed to analyze these immune cells for genetic and protein differences when compared to normal donor T cells freshly after isolation and 35 days after in vitro stimulation (IVS) with HLA-matched RCC tumor cells.
  
  
  METHODS
  We investigated gene expression profiles of tumor-reactive CD8(+) T cells obtained from RCC patient and compared with their HLA-matched healthy sibling donors using a microarray approach. In addition, miRNAs analysis was performed in a validation cohort of peripheral blood CD8(+) T cells from 25 RCC patients compared to 15 healthy volunteers.
  
  
  RESULTS
  We observed that CD8(+) T cells from RCC patients expressed reduced levels of anti-apoptotic and proliferation-associated gene products when compared with normal donor T cells both pre- and post-IVS. In particular, JAK3 and MCL-1 were down-regulated in patient CD8(+) T cells versus their normal counterparts, likely due to defective suppressor activity of miR-29b and miR-198 in RCC CD8(+) T cells. Indeed, specific inhibition of miR-29b or miR-198 in peripheral blood mononuclear cells (PBMCs) isolated from RCC patients, resulted in the up-regulation of JAK3 and MCL-1 proteins and significant improvement of cell survival in vitro.
  
  
  CONCLUSIONS
  Our results suggest that miR-29b and miR-198 dysregulation in RCC patient CD8(+) T cells is associated with dysfunctional immunity and foreshadow the development of miR-targeted therapeutics to correct such T cell defects in vivo.
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Margherita
authors.initials: M
authors.lastname: Gigante
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Pontrelli
authors.affiliation: Department of Medicine III, Hematology and Oncology, University Medical Center of Regensburg, Regensburg, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Herr
authors.affiliation: Department of Medical and Surgical Sciences, Faculty of Medicine, University of Foggia, Viale Luigi Pinto, 1, 71100, Foggia, Italy.
authors.forename: Maddalena
authors.initials: M
authors.lastname: Gigante
authors.affiliation: Department of Internal Medicine and Oncology, University of Bari, Bari, Italy.
authors.forename: Morena
authors.initials: M
authors.lastname: D'Avenia
authors.affiliation: Renal Unit, Department of Medicine, University of Verona, Verona, Italy.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Zaza
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Cavalcanti
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Accetturo
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Lucarelli
authors.affiliation: Department of Medical and Surgical Sciences, Faculty of Medicine, University of Foggia, Viale Luigi Pinto, 1, 71100, Foggia, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Carrieri
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Battaglia
authors.affiliation: Department of Dermatology and Immunology, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Storkus
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Loreto
authors.initials: L
authors.lastname: Gesualdo
authors.affiliation: Department of Medical and Surgical Sciences, Faculty of Medicine, University of Foggia, Viale Luigi Pinto, 1, 71100, Foggia, Italy. elena.ranieri@unifg.it.
authors.forename: Elena
authors.initials: E
authors.lastname: Ranieri
mesh_terms: D000328:Adult; D000368:Aged; D017209:Apoptosis / Q000235:genetics; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000276:immunology*; D002469:Cell Separation; D015536:Down-Regulation / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D053616:Janus Kinase 3 / Q000235:genetics / Q000378:metabolism*; D007680:Kidney Neoplasms / Q000235:genetics / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D014019:Tissue Donors; D014162:Transfection; D014184:Transplantation, Homologous; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000235:genetics
chemical_list: C579095:MCL1 protein, human; C535221:MIRN198 microRNA, human; C532045:MIRN29a microRNA, human; D035683:MicroRNAs; D064549:Myeloid Cell Leukemia Sequence 1 Protein; C507933:JAK3 protein, human; D053616:Janus Kinase 3
keywords: Apoptosis; CD8+ T cells; JAK3; MCL-1; Renal cell carcinoma; miRNA