Identification of a New Target of miR-16, Vacuolar Protein Sorting 4a

• DOI: 10.1371/journal.pone.0101509
• Publication Date: 2014-07-17T18:50:40Z
• Authors: Neeta Adhikari; Weihua Guan; Brian Capaldo; Aaron J. Mackey; Marjorie Carlson; Sundaram Ramakris...; Dinesha Walek; Manu Gupta; Adam Mitchell; Peter Eckman; Ranjit John; Euan Ashley; Paul J. Barton; Jennifer L. Hall
• Keywords: EXPRESSION; Male; 0301 basic medicine; 570; Vacuolar Proton-Translocating ATPases; General Science & Technology; Science; 610; Polymorphism, Single Nucleotide; CELL-PROLIFERATION; Cell Line; 03 medical and health sciences; Humans; Polymorphism; 3' Untranslated Regions; Aged; Heart Failure; Science & Technology; Binding Sites; Endosomal Sorting Complexes Required for Transport; Myocardium; Q; R; Single Nucleotide; Middle Aged; FAMILY; Multidisciplinary Sciences; MicroRNAs; HEK293 Cells; MICRORNA-16; DISEASES; OSTEOSARCOMA; Science & Technology - Other Topics; Medicine; ATPases Associated with Diverse Cellular Activities; Female; Heart-Assist Devices; Research Article

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).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).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).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.