Exome Sequencing Identifies a Mutation in EYA4 as a Novel Cause of Autosomal Dominant Non-Syndromic Hearing Loss - Fig 2

<p>(A) SH-01 audiometric phenotype. Pure-tone bone and air conduction thresholds as well as otoacoustic emission are presented for the right ears of the SH-01 family members. Two representative stages (early and late) are presented, with corresponding audiograms. Blue half-frame and open, red circles indicate the bone and air conduction thresholds, respectively, for the indicated ages, from youngest to oldest. Both conduction thresholds were consistent with predominant SNHL, and some individuals at the later stage displayed conductive hearing loss. In the family, Otoacoustic emission could be induced at most frequencies in individuals with normal hearing, while it was the reverse in affected individuals. (B, C) Based on the CT scan and MRI data of the proband (III9) and his aunt (II2), the mastoid process and cochlea were well developed, and the ossicular chain was intact. Additionally, both the internal auditory meatus and the membranous labyrinth were well developed.</p

Exome Sequencing Identifies a Mutation in EYA4 as a Novel Cause of Autosomal Dominant Non-Syndromic Hearing Loss - Fig 3

<p>(A) Read coverage of each chromosome and mutation loci statistical results of the patients and of the normal control; (B) DNA sequence chromatograms presenting the two heterozygous missense mutations c.511G>C; p.G171R in affected individuals (upper panel) compared with the wild-type controls (lower panel). (C) The structure of the EYA4 gene. The EYA4 gene has 20 exons. The c.511G>C; p.G171R mutation identified in the EYA4 gene is in exon 8. The conservation analysis indicates that the Gly residue at 171 in the EYA4 protein is not conserved across <i>Homo sapiens</i>, <i>Pan troglodytes</i>, <i>Macaca mulatta</i>, <i>Mus musculus</i>, <i>Gallus gallus</i>, <i>Takifugu rubripes</i>, <i>Danio rerio</i> and <i>Xenopus tropicalis</i>.</p

The effects of known EYA4 mutations on the EYA4 protein structure and on the cardiac phenotype.

<p>The number of amino acids of each allele product is indicated. Mutations that truncate the C-terminal Eya domain are associated with DFNA10 hearing loss and with a normal cardiac phenotype, whereas E193 truncates the N-terminal variable region and results in hearing loss plus dilated cardiomyopathy. Asp171 is in the variable region domain, and patients with this mutation presented with hearing loss and a normal cardiac phenotype.</p

Partial pedigree of the Chinese family with nonsyndromic autosomal dominant SNHL.

<p>Open symbols denote unaffected individuals; filled black symbols denote affected individuals. The arrow labels the proband.</p

Summary of the audiological features of affected members of family SH-01.

<p>The degree of hearing loss was defined according to pure-tone averages (PTA), which were based on the following four frequencies: 0.5, 1, 2 and 4 kHz.</p><p>Summary of the audiological features of affected members of family SH-01.</p

The effect of pre-eclampsia-like syndrome induced by L-NAME on learning and memory and hippocampal glucocorticoid receptor expression: A rat model

<p><i>Objective</i>: We aimed to study the impacts of pre-eclampsia on the cognitive and learning capabilities of adolescent rat offspring and to explore the possible underlying mechanisms at the molecular level. <i>Methods</i>: Pregnant rats were subcutaneously injected with saline solution (control) (<i>n</i> = 16) or NG-nitro-L-arginine methyl ester (L-NAME) (<i>n</i> = 16) from the 13th day of gestation until parturition. The brain tissues from fetal rats delivered by cesarean section were examined in both groups with hematoxylin and eosin (H&E) staining. Rats born vaginally in both groups were subjected to the Morris water maze test when 8-week-old and their hippocampi were analyzed for glucocorticoid receptor (GR) expression. <i>Results</i>: A pre-eclampsia-like model was successfully built in pregnant rats by infusion of the NO synthase inhibitor L-NAME, including phenotypes as maternal hypertension and proteinuria, high stillbirth rate, and fetal growth retardation. Neuroepithelial cell proliferation was found in the hippocampus of fetal rats in the L-NAME group. Grown to 8-week-old, the L-NAME group showed significantly longer escape latency than the control group in the beginning as well as in the end of navigation trials. At the same time, the swimming distance achieved by the L-NAME group was significantly longer than that of the control group. Such differences in cognitive and learning capabilities between the two groups were not gender dependent. Besides, the 8-week-old rats in the L-NAME group had increased GR expression in the hippocampus than the control group. <i>Conclusion</i>: Pre-eclampsia would impair cognitive and learning capabilities in adolescent offspring, and the upregulated expression of hippocampal GR may be involved in the underlying mechanisms.</p

Video_1_miR-210-3p protects against osteoarthritis through inhibiting subchondral angiogenesis by targeting the expression of TGFBR1 and ID4.mp4

Excessive subchondral angiogenesis is a key pathological feature of osteoarthritis (OA), as it alters the balance of subchondral bone remodeling and causes progressive cartilage degradation. We previously found that miR-210-3p correlates negatively with angiogenesis, though the specific mechanism of miR-210-3p-related angiogenesis in subchondral bone during OA progression remains unclear. This study was conducted to identify the miR-210-3p-modulating subchondral angiogenesis mechanism in OA and investigate its therapeutic effect. We found that miR-210-3p expression correlated negatively with subchondral endomucin positive (Emcn+) vasculature in the knee joints of OA mice. miR-210-3p overexpression regulated the angiogenic ability of endothelial cells (ECs) under hypoxic conditions in vitro. Mechanistically, miR-210-3p inhibited ECs angiogenesis by suppressing transforming growth factor beta receptor 1 (TGFBR1) mRNA translation and degrading DNA-binding inhibitor 4 (ID4) mRNA. In addition, TGFBR1 downregulated the expression of ID4. Reduced ID4 levels led to a negative feedback regulation of TGFBR1, enhancing the inhibitory effect of miR-210-3p on angiogenesis. In OA mice, miR-210-3p overexpression in ECs via adeno-associated virus (AAV) alleviated cartilage degradation, suppressed the type 17 immune response and relieved symptoms by attenuating subchondral Emcn+ vasculature and subchondral bone remodeling. In conclusion, we identified a miR-210-3p/TGFBR1/ID4 axis in subchondral ECs that modulates OA progression via subchondral angiogenesis, representing a potential OA therapy target.</p

DataSheet_1_miR-210-3p protects against osteoarthritis through inhibiting subchondral angiogenesis by targeting the expression of TGFBR1 and ID4.docx

Excessive subchondral angiogenesis is a key pathological feature of osteoarthritis (OA), as it alters the balance of subchondral bone remodeling and causes progressive cartilage degradation. We previously found that miR-210-3p correlates negatively with angiogenesis, though the specific mechanism of miR-210-3p-related angiogenesis in subchondral bone during OA progression remains unclear. This study was conducted to identify the miR-210-3p-modulating subchondral angiogenesis mechanism in OA and investigate its therapeutic effect. We found that miR-210-3p expression correlated negatively with subchondral endomucin positive (Emcn+) vasculature in the knee joints of OA mice. miR-210-3p overexpression regulated the angiogenic ability of endothelial cells (ECs) under hypoxic conditions in vitro. Mechanistically, miR-210-3p inhibited ECs angiogenesis by suppressing transforming growth factor beta receptor 1 (TGFBR1) mRNA translation and degrading DNA-binding inhibitor 4 (ID4) mRNA. In addition, TGFBR1 downregulated the expression of ID4. Reduced ID4 levels led to a negative feedback regulation of TGFBR1, enhancing the inhibitory effect of miR-210-3p on angiogenesis. In OA mice, miR-210-3p overexpression in ECs via adeno-associated virus (AAV) alleviated cartilage degradation, suppressed the type 17 immune response and relieved symptoms by attenuating subchondral Emcn+ vasculature and subchondral bone remodeling. In conclusion, we identified a miR-210-3p/TGFBR1/ID4 axis in subchondral ECs that modulates OA progression via subchondral angiogenesis, representing a potential OA therapy target.</p

The results of 55 patients detected by MLPA.

<p>A total of 43 cases (78.2%) showed 22q11.2 heterozygous deletion, of whom 40 (93.0%) exhibited typical 3-Mb deletion, while 3 (7.0%) showed proximal 1.5-Mb deletion; no case was found having atypical deletion on 22q11.2. Only 1 case (1.8%) had 3-Mb duplication. None of the chromosomal abnormalities in the MLPA kit were found in the other 11 patients.</p

Characteristic face of VCFS.

<p>These Chinese VCFS patients all presented with a characteristic face, consisting of vertically long face, narrow palpebral fissures, fleshy nose with a broad nasal root, flattened malar region, retrognathia, and sometimes overfolded helix (E) or cup-shaped ear (F).</p