FERNANDO PÉREZ SANZ¹, ÁNGEL ESTEBAN GIL², MARÍA ANTONIA PARREÑO GONZÁLEZ¹, MARÍA DEL CARMEN LEGAZ GARCÍA², ANA ISABEL ANTON GARCIA³, VERÓNICA PALMA BARQUEROS⁴, NATALIYA BOHDAN ⁵, JUAN FRANCISCO RUIZ PIVIDAL⁶, Vicente Vicente García⁷, Loredana Bury⁸, José María Bastida⁹, Mª Luisa LOZANO ALMELA⁷, José Rivera Pozo⁷

(1) PLATAFORMA DE INFORMÁTICA BIOMÉDICA Y BIOINFORMÁTICA, IMIB, España
(2) TECNOLOGÍAS DE MODELADO, PROCESAMIENTO Y GESTIÓN DEL CONOCIMIENTO, IMIB, España
(3) PLATAFORMA DE GENÓMICA, IMIB, España
(4) Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Arrixaca, U765-CIBERER, Murcia, Spain, 30003, España (Región de Murcia)
(5)
(6) Investigador, 30800, España (Región de Murcia)
(7) HEMATOLOGÍA Y ONCOLOGÍA MÉDICA CLÍNICO-EXPERIMENTAL, IMIB, España
(8) Department of Medicine, Section of Internal and Cardiovascular Medicine, University of Perugia, Perugia, Italia
(9) Unidad de Trombosis y Hemostasia. H. Universitario de Salamanca-IBSAL, Salamanca, España (Castilla y León)

High-throughput sequencing (HTS) is being introduced into diagnosis practice in inherited platelet disorders (IPDs). A major difficulty in this setting is analysis of the massive molecular information obtained, requiring bioinformatic expertise. The aim of this study is the development of a web platform allowing comfortable analysis of molecular data from HTS investigation of IPDs.

DNA from IPD patients were assayed with Ion Torrent platform, with probes targeting exons, UTR and flanking regions of 90 genes (241,348 bp). Sequence data are formatted in FASTA files and aligned against hg19 in BAM files. Then, variant call format (VCF) files listing all gene sequence variations are created, and uploaded in cluster servers. DIGEVAR “Discovering Genetic Variants” web platform was developed in JAVA to access VCF files (https://digevar.imib.es). DIGEVAR allows authorized users to perform multiple selection of variant filtering strategies including: a) variant severity; b) variant gene location; c) minor allele frequency (MAF); d) sequence quality and coverage; f) gene (single or multiple analysis); g) DNA sample(s). By executing “Filter” DIGEVAR provides the list of variants fulfilling the filtering criteria.

So far, 151 VCF files listing 5685 different variants, are available for DIGEVAR analysis. Platform connection with public databases (ENSEMBL, NCBI, CLINVAR, ExAC,etc.) and variant analysis software (MutationTaster, Polyphen, Sift, PDB, etc.), provides extensive information including: allele frequency, rs ID, transcript, nucleotide/protein change, variant consequence in gene/protein, phylogenetics, CLINVAR significance. Complementary modules in DIGEVAR provide: i) coverage information for each explored region in each gene; ii) variant information in public databases (PubMed, dbSNP, etc.); iii) variant interpretation according to ACMG and AMP guidelines.

DIGEVAR is a user-friendly web platform allowing a rapid analysis, by non- experts in bioinformatics, of data from multigenic HTS in IPDs, or in other diseases. DIGEVAR facilitates the analysis of HTS data in clinical practice (ISCIII&Feder 17/01311&17/01966&CB15/0055; FS-19873/GERM/15; GRS-1647/A/17V; IBSAL-IBY17/00006).