- Open Access
miRdSNP: a database of disease-associated SNPs and microRNA target sites on 3'UTRs of human genes
© Bruno et al; licensee BioMed Central Ltd. 2012
- Received: 22 September 2011
- Accepted: 25 January 2012
- Published: 25 January 2012
Single nucleotide polymorphisms (SNPs) can lead to the susceptibility and onset of diseases through their effects on gene expression at the posttranscriptional level. Recent findings indicate that SNPs could create, destroy, or modify the efficiency of miRNA binding to the 3'UTR of a gene, resulting in gene dysregulation. With the rapidly growing number of published disease-associated SNPs (dSNPs), there is a strong need for resources specifically recording dSNPs on the 3'UTRs and their nucleotide distance from miRNA target sites. We present here miRdSNP, a database incorporating three important areas of dSNPs, miRNA target sites, and diseases.
miRdSNP provides a unique database of dSNPs on the 3'UTRs of human genes manually curated from PubMed. The current release includes 786 dSNP-disease associations for 630 unique dSNPs and 204 disease types. miRdSNP annotates genes with experimentally confirmed targeting by miRNAs and indexes miRNA target sites predicted by TargetScan and PicTar as well as potential miRNA target sites newly generated by dSNPs. A robust web interface and search tools are provided for studying the proximity of miRNA binding sites to dSNPs in relation to human diseases. Searches can be dynamically filtered by gene name, miRBase ID, target prediction algorithm, disease, and any nucleotide distance between dSNPs and miRNA target sites. Results can be viewed at the sequence level showing the annotated locations for miRNA target sites and dSNPs on the entire 3'UTR sequences. The integration of dSNPs with the UCSC Genome browser is also supported.
miRdSNP provides a comprehensive data source of dSNPs and robust tools for exploring their distance from miRNA target sites on the 3'UTRs of human genes. miRdSNP enables researchers to further explore the molecular mechanism of gene dysregulation for dSNPs at posttranscriptional level. miRdSNP is freely available on the web at http://mirdsnp.ccr.buffalo.edu.
- miRNA Target
- UCSC Genome Browser
- miRNA Binding Site
- miRNA Target Site
- miRNA Target Prediction
Single nucleotide polymorphisms (SNPs) underlie disease susceptibility through their effects on protein function and gene expression. Most identified mutations are non-synonymous SNPs that result in amino acid changes in proteins. It is well known that non-coding disease-associated SNPs (dSNPs) within regulatory regions of the genome can result in gene dysregulation at either transcriptional or posttranscriptional level. One potential source for the latter is SNPs which create, destroy, or modify the efficiency of miRNA binding to the 3'UTR of a gene. Supporting this idea, SNPs within the miRNA target sites of genes have been implicated in hippocampal sclerosis , parkinson disease , tourette's syndrome , asthma , cardiovascular disease , neurodegenerative disease , periodontal diseases , tumor susceptibility , and various types of cancers [9–12]. Other than SNPs within miRNA target sites, SNPs outside miRNA binding site can affect miRNA function. One recent finding  demonstrates that a polymorphism outside the miR-24 binding site in the 3'UTR of human dihydrofolate reductase (DHFR) affects DHFR expression by interfering with miR-24 function, resulting in DHFR over-expression and methotrexate resistance. There is also a report suggesting that SNPs within a certain region on both sides of miRNA target sites have the highest influence on miRNA binding to the target sites and that SNPs on the rest of 3'UTR sequences have impact on miRNA function as well .
A few databases have been built to aid researchers in exploring the impact of SNPs on the binding of miRNA and targets. While polymiRTS  represents the polymorphism in putative miRNA target sites and their involvement in quantitative trait locus effects, Patrocles database compiles DNA sequence polymorphisms in the 3'UTR of genes in seven vertebrate species that perturb miRNA-mediated gene regulation . The findings of dSNPs on the 3'UTRs have been growing rapidly during the past few years. Furthermore, a few dSNPs [1–5, 7–12] have been proven to alter gene expression through modifying specific miRNA target sites, however the molecular mechanism causing diseases for majority of dSNPs on the 3'UTRs is largely not known. There is a strong need to have a database specifically recording dSNPs and tools for capturing their proximity to miRNA target sites on the 3'UTRs so that researchers can explore further the molecular mechanism of gene dysregulation for dSNPs at posttranscriptional level.
Aiming to provide a comprehensive data source of dSNPs affecting posttranscriptional regulation of disease-related genes and tools for exploring the nucleotide distance between miRNA target sites and dSNPs, we present here miRdSNP, a database of manually curated dSNPs on the 3'UTRs of human genes from available publications in PubMed. A robust web interface and advanced search tools are provided showing the nucleotide distance between dSNPs and predicted miRNA target sites from the most popular algorithms, namely TargetScan  and PicTar . We also incorporated all SNPs on 3'UTRs of individual genes into the database so the relationship of SNPs with both dSNPs and miRNA binding sites can be analyzed using the web interface. In addition, we also include predicted miRNA target sites generated by dSNPs based on our analysis and annotate genes with experimentally confirmed targeting by miRNAs from four separate curated databases.
We obtained 3'UTRs of human RefSeq genes (hg18 March 2006 assembly) from the UCSC Genome browser . A total of 19,834 genes (including introns) were parsed and loaded into miRdSNP and the chromosomal coordinates for each gene were indexed along with the exon lengths. If a gene had multiple transcripts we selected the one with the longest sequence length. We obtained the SNP dataset from UCSC Genome browser (NCBI dbSNP  Build 130). Of 18,833,531 SNPs we indexed the genomic coordinates for a subset of 175,351 located on 3'UTRs of 16,810 genes. SNPs aligning to more than 1 locus or mapped to intron regions were excluded. We then annotated dSNPs using an in-house developed data pipeline which searches for PubMed articles linked to SNPs. The data pipeline queries ELink from the Entrez Programming Utilities to find all PubMed IDs linked to SNPs via the "snp pubmed" link. We queried all 3'UTR SNPs and found 2,785 PubMed publications linked to 16,447 unique SNPs. We then manually reviewed these literatures and identified 630 dSNPs for 204 human diseases from 754 publications. The data pipeline for harvesting PubMed-SNP associations from Entrez is automated and the results are displayed in a web interface, allowing multiple users to manually review articles in parallel. This enables us to update the curated dSNP dataset frequently as the literature evolves.
We captured linkage disequilibrium (LD) information for each dSNP using the latest data provided by the HapMap project  (version 2009-04_rel27). We downloaded the raw LD files for each population and searched for pairs of genetic variants that included dSNPs. We then indexed all variants in strong LD of each dSNP using an R2 ≥ 0.80 threshold. Regional LD plots were also generated for each dSNP using a modified version of the R script provided by SNAP  and data from the CEU (CEPH Utah Residents with Northern and Western European Ancestry) population.
We obtained miRNA target site datasets from two miRNA target prediction algorithms, namely TargetScan 5.1 and PicTar which were found to have the highest precision and sensitivity out of eight commonly used algorithms . The data for each prediction algorithm was downloaded from the respective source, and the genomic coordinates for each target site were indexed and mapped to RefSeq genes. In addition, each miRNA target site was cross referenced with miRBase  and targets referencing dead or non-existent miRNAs were excluded. The genomic coordinates from PicTar were converted from hg17 assembly to hg18 assembly using the LiftOver utility from UCSC. The exon index for each miRNA target site was computed and used to calculate the nucleotide distance between SNPs and predicted miRNA target sites. This distance is calculated from the start or end location of the miRNA target "seed" region (~7nt), depending on whether the SNP is upstream or downstream of the miRNA target site. SNPs which fall inside the miRNA target "seed" region have a distance of 0. To address the low prediction specificity of the miRNA target prediction algorithms we incorporate data from four curation databases (TarBase , miRTarBase , miRecords , and miR2disease ) which collect experimentally confirmed miRNA target interactions. Genes with experimentally confirmed targeting by miRNAs were annotated in miRdSNP and displayed with a green check icon within the user interface.
To predict new miRNA target sites created by dSNPs, we first generated candidate sequences using a 6-nt flank up/down stream from each dSNP location replacing the dSNP with the observed allele. Using these new candidate sequences we searched for perfect match 7-mer seed regions from miRBase mature sequence data. We then extracted 25-nt flank upstream of the matching seed region and used the miRNA target prediction program miRanda  with default cutoff to further eliminate false positives. We were able to identify 180 newly created miRNA target sites from 138 dSNPs.
Loading and indexing new data in miRdSNP is an automated process allowing for streamlined updates to the database as new RefSeq, miRNA target prediction, and SNP data become available. All data in miRdSNP is available for download in raw text format (CSV, BED) with access to previous versions.
Utility and discussion
miRdSNP is an ongoing effort to create a comprehensive data source for exploring the effect of SNPs on miRNA binding in relation to human diseases. We are working on importing data from other miRNA target prediction algorithms such as DIANA-microT v3.0  and ElMMo . Since the accuracy of the manually curated database of dSNPs is an integral part of miRdSNP, we aim to further broaden the amount of data captured from the manual process. Data such as study design, sample size, and p-values would further enhance the ability to determine the disease-SNP association. We aim to update the dSNP curation database yearly and as new versions of the miRNA target prediction algorithms become available.
miRdSNP is freely available on the web at http://mirdsnp.ccr.buffalo.edu.
This work was supported, in part, by United States Public Health Service (National Institutes of Health) grant 1R01EY020545 (ZH) and by an unrestricted grant from Research to Prevent Blindness (ZH).
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