title: Identification of a new target of miR-16, Vacuolar Protein Sorting 4a.
abstract: BACKGROUND
  The rationale was to utilize a bioinformatics approach to identify miRNA binding sites in genes with single nucleotide mutations (SNPs) to discover pathways in heart failure (HF).
  
  
  OBJECTIVE
  The objective was to focus on the genes containing miRNA binding sites with miRNAs that were significantly altered in end-stage HF and in response to a left ventricular assist device (LVAD).
  
  
  RESULTS
  BEDTools v2.14.3 was used to discriminate SNPs within predicted 3'UTR miRNA binding sites. A member of the miR-15/107 family, miR-16, was decreased in the circulation of end-stage HF patients and increased in response to a LVAD (p<0.001). MiR-16 decreased Vacuolar Protein Sorting 4a (VPS4a) expression in HEK 293T cells (p<0.01). The SNP rs16958754 was identified in the miR-15/107 family binding site of VPS4a which abolished direct binding of miR-16 to the 3'UTR of VPS4a (p<0.05). VPS4a was increased in the circulation of end-stage HF patients (p<0.001), and led to a decrease in the number of HEK 293T cells in vitro (p<0.001).
  
  
  CONCLUSIONS
  We provide evidence that miR-16 decreases in the circulation of end-stage HF patients and increases with a LVAD. Modeling studies suggest that miR-16 binds to and decreases expression of VPS4a. Overexpression of VPS4a decreases cell number. Together, these experiments suggest that miR-16 and VPS4a expression are altered in end-stage HF and in response to unloading with a LVAD. This signaling pathway may lead to reduced circulating cell number in HF.
authors.affiliation: Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Neeta
authors.initials: N
authors.lastname: Adhikari
authors.affiliation: Division of Biostatistics, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Weihua
authors.initials: W
authors.lastname: Guan
authors.affiliation: Center for Public Health Genomics, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: Brian
authors.initials: B
authors.lastname: Capaldo
authors.affiliation: Center for Public Health Genomics, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Mackey
authors.affiliation: Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Marjorie
authors.initials: M
authors.lastname: Carlson
authors.affiliation: Department of Pharmacology, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Sundaram
authors.initials: S
authors.lastname: Ramakrishnan
authors.affiliation: University of Minnesota Genomics Center, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Dinesha
authors.initials: D
authors.lastname: Walek
authors.affiliation: National Institute of Heart Research, Cardiovascular Biomedical Research Unit, Royal Brompton and Harefield Trust, London, United Kingdom.
authors.forename: Manu
authors.initials: M
authors.lastname: Gupta
authors.affiliation: Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Adam
authors.initials: A
authors.lastname: Mitchell
authors.affiliation: Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Peter
authors.initials: P
authors.lastname: Eckman
authors.affiliation: Department of Surgery, Cardiothoracic Surgery Division, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Ranjit
authors.initials: R
authors.lastname: John
authors.affiliation: Center for Inherited Cardiovascular Disease, Department of Medicine, Stanford University, Stanford, California, United States of America.
authors.forename: Euan
authors.initials: E
authors.lastname: Ashley
authors.affiliation: Heart Science Centre, National Heart and Lung Institute, Imperial College, London, United Kingdom.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Barton
authors.affiliation: Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Hall
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D000074183:ATPases Associated with Diverse Cellular Activities; D000368:Aged; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D056827:Endosomal Sorting Complexes Required for Transport / Q000096:biosynthesis / Q000097:blood / Q000235:genetics*; D005260:Female; D057809:HEK293 Cells; D006333:Heart Failure / Q000097:blood / Q000235:genetics* / Q000628:therapy; D006353:Heart-Assist Devices; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000097:blood / Q000235:genetics*; D008875:Middle Aged; D009206:Myocardium / Q000473:pathology; D020641:Polymorphism, Single Nucleotide; D025262:Vacuolar Proton-Translocating ATPases / Q000096:biosynthesis / Q000097:blood / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; D056827:Endosomal Sorting Complexes Required for Transport; C505362:MIRN16 microRNA, human; D035683:MicroRNAs; D025262:Vacuolar Proton-Translocating ATPases; D000074183:ATPases Associated with Diverse Cellular Activities; C490305:VPS4A protein, human