rine MicroRNA in T1D Albuminuric vs Normoalbuminuric in the MA group Pathway Signal Transduction Signaling by SCF-KIT Signaling by Insulin receptor Signaling by NGF Signaling by Rho GTPases Signaling by ERBB4 Signaling by ERBB2 Signaling by PDGF Signaling by VEGF Signaling by EGFR Dowstream signaling of activated FGFR Signaling by BMP Signaling by TGFb DAG and IP3 signaling PIP3 activates AKT signaling RAF/MAP kinase cascade Signaling by Notch Interaction of integrin a5b3 with JNJ-26481585 manufacturer fibrillin Interaction of integrin a5b3 with von Willbrand factor Integrin cell surface interactions Cell-Cell Communication Cell Cycle G0 and early G1 Metabolism Metabolism of lipids and lipoproteins Cysteine formation from homocysteine Integration of energy metabolism Metabolism of proteins Post-translational protein modification Transmembrane transport of small molecules Membrane trafficking Apoptosis Caspase-8 is formed from procaspase-8 Gene Expression RNA Polymerase 25136132 II Transcription Capping complex formation Nuclear Receptor Transcription Steroid hormones Vitamin D metabolism Activated AMPK stimulates fatty-acid oxidation in muscle Neuronal System Heterodimerization of CEACAMs Transmission across Chemical Synapses Immune System Interleukin-2 signaling 14-3-3 zeta binding allows recruitment of PI3K Signaling by interleukins Urine MicroRNA in T1D Albuminuric vs Normoalbuminuric in the MA group Pathway Hemostasis Platelet homeostasis Platelet activation, signaling and aggregation 0.008 0.011 14/56 35/187 P-value Fraction Overt vs Normal P-value,0.001 Fraction 206/426 P-value: the p-value of the hypergeometric test unadjusted for multiple comparisons, Fraction: number of proteins in the pathway that are targets of differentially expressed miRNAs over the total number of proteins in each 
pathway. doi:10.1371/journal.pone.0054662.t005 precluding a meaningful assessment with this structured vocabulary. The results from Gene Ontology analysis were consistent with REACTOME, and also identified enrichment of terms relating to nitrogen compound metabolism, Golgi/membrane/ER vesicle recycling, ubiquitin-dependent degradation, cell adhesion and cell adhesion. In addition, GO analyses also suggested the enrichment of renal and non-renal developmental pathways, myoblast determination innate immunity and free radical generation/oxidative stress. Discussion In this paper we report the changes of urinary miRNA spectrum in T1D patients with different stages of albuminuria and nephropathy. We found concentration changes on specific miRNAs that may involve in specific pathways known to be altered in various forms of renal diseases. Since the kidney is the most likely 23388095 source of these urinary miRNAs, we suggest that these miRNAs may be of biological and clinical significance in T1D. A global Principal Component Analysis viewpoint of the microRNA profiles analyzed in this report suggests that there are some differences in the expression of urinary microRNA which appear to follow the clinical classification of patients and urinary samples with respect to albumin excretion. The apparent clustering of profiles from patients who had been matched into pairs, suggests that there are other factors affecting urinary microRNA besides the clinical classification of disease. Such factors are likely related to the variables we used in patient matching e.g. age, sex, and duration of disease and level of glycemic control. This observation justifies post-hoc our decision to explore specific microR
