TY - JOUR T1 - Next-generation sequencing identified SPATC1L as a possible candidate gene for both early-onset and age-related hearing loss. JF - Eur J Hum Genet Y1 - 2019 A1 - Morgan, Anna A1 - Vuckovic, Dragana A1 - Krishnamoorthy, Navaneethakrishnan A1 - Rubinato, Elisa A1 - Ambrosetti, Umberto A1 - Castorina, Pierangela A1 - Franzè, Annamaria A1 - Vozzi, Diego A1 - La Bianca, Martina A1 - Cappellani, Stefania A1 - Di Stazio, Mariateresa A1 - Gasparini, Paolo A1 - Girotto, Giorgia AB -

Hereditary hearing loss (HHL) and age-related hearing loss (ARHL) are two major sensory diseases affecting millions of people worldwide. Despite many efforts, additional HHL-genes and ARHL genetic risk factors still need to be identified. To fill this gap a large genomic screening based on next-generation sequencing technologies was performed. Whole exome sequencing in a 3-generation Italian HHL family and targeted re-sequencing in 464 ARHL patients were performed. We detected three variants in SPATC1L: a nonsense allele in an HHL family and a frameshift insertion and a missense variation in two unrelated ARHL patients. In silico molecular modelling of all variants suggested a significant impact on the structural stability of the protein itself, likely leading to deleterious effects and resulting in truncated isoforms. After demonstrating Spatc1l expression in mice inner ear, in vitro functional experiments were performed confirming the results of the molecular modelling studies. Finally, a candidate-gene population-based statistical study in cohorts from Caucasus and Central Asia revealed a statistically significant association of SPATC1L with normal hearing function at low and medium hearing frequencies. Overall, the amount of different genetic data presented here (variants with early-onset and late-onset hearing loss in addition to genetic association with normal hearing function), together with relevant functional evidence, likely suggest a role of SPATC1L in hearing function and loss.

VL - 27 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/30177775?dopt=Abstract ER - TY - JOUR T1 - Genome-wide association analysis on normal hearing function identifies PCDH20 and SLC28A3 as candidates for hearing function and loss. JF - Hum Mol Genet Y1 - 2015 A1 - Vuckovic, Dragana A1 - Dawson, Sally A1 - Scheffer, Deborah I A1 - Rantanen, Taina A1 - Morgan, Anna A1 - Di Stazio, Mariateresa A1 - Vozzi, Diego A1 - Nutile, Teresa A1 - Concas, Maria P A1 - Biino, Ginevra A1 - Nolan, Lisa A1 - Bahl, Aileen A1 - Loukola, Anu A1 - Viljanen, Anne A1 - Davis, Adrian A1 - Ciullo, Marina A1 - Corey, David P A1 - Pirastu, Mario A1 - Gasparini, Paolo A1 - Girotto, Giorgia AB -

Hearing loss and individual differences in normal hearing both have a substantial genetic basis. Although many new genes contributing to deafness have been identified, very little is known about genes/variants modulating the normal range of hearing ability. To fill this gap, we performed a two-stage meta-analysis on hearing thresholds (tested at 0.25, 0.5, 1, 2, 4, 8 kHz) and on pure-tone averages (low-, medium- and high-frequency thresholds grouped) in several isolated populations from Italy and Central Asia (total N = 2636). Here, we detected two genome-wide significant loci close to PCDH20 and SLC28A3 (top hits: rs78043697, P = 4.71E-10 and rs7032430, P = 2.39E-09, respectively). For both loci, we sought replication in two independent cohorts: B58C from the UK (N = 5892) and FITSA from Finland (N = 270). Both loci were successfully replicated at a nominal level of significance (P < 0.05). In order to confirm our quantitative findings, we carried out RT-PCR and reported RNA-Seq data, which showed that both genes are expressed in mouse inner ear, especially in hair cells, further suggesting them as good candidates for modulatory genes in the auditory system. Sequencing data revealed no functional variants in the coding region of PCDH20 or SLC28A3, suggesting that variation in regulatory sequences may affect expression. Overall, these results contribute to a better understanding of the complex mechanisms underlying human hearing function.

VL - 24 IS - 19 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26188009?dopt=Abstract ER - TY - JOUR T1 - PSIP1/LEDGF: a new gene likely involved in sensorineural progressive hearing loss. JF - Sci Rep Y1 - 2015 A1 - Girotto, Giorgia A1 - Scheffer, Deborah I A1 - Morgan, Anna A1 - Vozzi, Diego A1 - Rubinato, Elisa A1 - Di Stazio, Mariateresa A1 - Muzzi, Enrico A1 - Pensiero, Stefano A1 - Giersch, Anne B A1 - Corey, David P A1 - Gasparini, Paolo AB -

Hereditary Hearing Loss (HHL) is an extremely heterogeneous disorder. Approximately 30 out of 80 known HHL genes are associated with autosomal dominant forms. Here, we identified PSIP1/LEDGF (isoform p75) as a novel strong candidate gene involved in dominant HHL. Using exome sequencing we found a frameshift deletion (c.1554_1555del leading to p.E518Dfs*2) in an Italian pedigree affected by sensorineural mild-to-moderate HHL but also showing a variable eye phenotype (i.e. uveitis, optic neuropathy). This deletion led to a premature stop codon (p.T519X) with truncation of the last 12 amino acids. PSIP1 was recently described as a transcriptional co-activator regulated by miR-135b in vestibular hair cells of the mouse inner ear as well as a possible protector against photoreceptor degeneration. Here, we demonstrate that it is ubiquitously expressed in the mouse inner ear. The PSIP1 mutation is associated with a peculiar audiometric slope toward the high frequencies. These findings indicate that PSIP1 likely plays an important role in HHL.

VL - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26689366?dopt=Abstract ER - TY - JOUR T1 - Clinical and genetic aspects of Bernard-Soulier syndrome: searching for genotype/phenotype correlations. JF - Haematologica Y1 - 2011 A1 - Savoia, Anna A1 - Pastore, Annalisa A1 - De Rocco, Daniela A1 - Civaschi, Elisa A1 - Di Stazio, Mariateresa A1 - Bottega, Roberta A1 - Melazzini, Federica A1 - Bozzi, Valeria A1 - Pecci, Alessandro A1 - Magrin, Silvana A1 - Balduini, Carlo L A1 - Noris, Patrizia KW - Adolescent KW - Adult KW - Amino Acid Sequence KW - Bernard-Soulier Syndrome KW - Blood Platelets KW - Cell Shape KW - Child KW - Child, Preschool KW - Female KW - Genetic Association Studies KW - Genetic Markers KW - Hemorrhage KW - Homozygote KW - Humans KW - Italy KW - Male KW - Membrane Glycoproteins KW - Middle Aged KW - Molecular Sequence Data KW - Platelet Aggregation KW - Platelet Count KW - Platelet Glycoprotein GPIb-IX Complex KW - Point Mutation KW - Polymerase Chain Reaction KW - Ristocetin KW - Thrombocytopenia KW - von Willebrand Factor KW - Young Adult AB -

BACKGROUND: Bernard-Soulier syndrome is a severe bleeding disease due to a defect of GPIb/IX/V, a platelet complex that binds the von Willebrand factor. Due to the rarity of the disease, there are reports only on a few cases compromising any attempt to establish correlations between genotype and phenotype. In order to identify any associations, we describe the largest case series ever reported, which was evaluated systematically at the same center.

DESIGN AND METHODS: Thirteen patients with the disease and seven obligate carriers were enrolled. We collected clinical aspects and determined platelet features, including number and size, expression of membrane glycoproteins, and ristocetin induced platelet aggregation. Mutations were identified by direct sequencing of the GP1BA, GP1BB, and GP9 genes and their effect was shown by molecular modeling analyses.

RESULTS: Patients all had a moderate thrombocytopenia with giant platelets and a bleeding tendency whose severity varied among individuals. Consistent with expression levels of GPIbα always lower than 10% of control values, platelet aggregation was absent or severely reduced. Homozygous mutations were identified in the GP1BA, GP1BB and GP9 genes; six were novel alterations expected to destabilize the conformation of the respective protein. Except for obligate carriers of a GP9 mutation with a reduced GPIb/IX/V expression and defective aggregation, all the other carriers had no obvious anomalies.

CONCLUSIONS: Regardless of mutations identified, the patients' bleeding diathesis did not correlate with thrombocytopenia, which was always moderate, and platelet GPIbα expression, which was always severely impaired. Obligate carriers had features similar to controls though their GPIb/IX/V expression showed discrepancies. Aware of the limitations of our cohort, we cannot define any correlations. However, further investigations should be encouraged to better understand the causes of this rare and underestimated disease.

VL - 96 IS - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21173099?dopt=Abstract ER - TY - JOUR T1 - Mutations in the 5' UTR of ANKRD26, the ankirin repeat domain 26 gene, cause an autosomal-dominant form of inherited thrombocytopenia, THC2. JF - Am J Hum Genet Y1 - 2011 A1 - Pippucci, Tommaso A1 - Savoia, Anna A1 - Perrotta, Silverio A1 - Pujol-Moix, Núria A1 - Noris, Patrizia A1 - Castegnaro, Giovanni A1 - Pecci, Alessandro A1 - Gnan, Chiara A1 - Punzo, Francesca A1 - Marconi, Caterina A1 - Gherardi, Samuele A1 - Loffredo, Giuseppe A1 - De Rocco, Daniela A1 - Scianguetta, Saverio A1 - Barozzi, Serena A1 - Magini, Pamela A1 - Bozzi, Valeria A1 - Dezzani, Luca A1 - Di Stazio, Mariateresa A1 - Ferraro, Marcella A1 - Perini, Giovanni A1 - Seri, Marco A1 - Balduini, Carlo L KW - Ankyrin Repeat KW - Base Sequence KW - Chromosome Breakage KW - Chromosome Disorders KW - Conserved Sequence KW - Female KW - Genes, Dominant KW - Genetic Loci KW - Haploinsufficiency KW - Humans KW - Male KW - Molecular Sequence Data KW - Mutation KW - Pedigree KW - Thrombocytopenia AB -

THC2, an autosomal-dominant thrombocytopenia described so far in only two families, has been ascribed to mutations in MASTL or ACBD5. Here, we show that ANKRD26, another gene within the THC2 locus, and neither MASTL nor ACBD5, is mutated in eight unrelated families. ANKRD26 was also found to be mutated in the family previously reported to have an ACBD5 mutation. We identified six different ANKRD26 mutations, which were clustered in a highly conserved 19 bp sequence located in the 5' untranslated region. Mutations were not detected in 500 controls and are absent from the 1000 Genomes database. Available data from an animal model and Dr. Watson's genome give evidence against haploinsufficiency as the pathogenetic mechanism for ANKRD26-mediated thrombocytopenia. The luciferase reporter assay suggests that these 5' UTR mutations might enhance ANKRD26 expression. ANKRD26 is the ancestor of a family of primate-specific genes termed POTE, which have been recently identified as a family of proapoptotic proteins. Dysregulation of apoptosis might therefore be the pathogenetic mechanism, as demonstrated for another thrombocytopenia, THC4. Further investigation is needed to provide evidence supporting this hypothesis.

VL - 88 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21211618?dopt=Abstract ER -