ARHI Manhattan plot in GERA non-Hispanic whites.

<p>SNPs meeting genome-wide significance (p<5x10^-8) are above the red line. The circles indicate genotyped SNPs, and the triangles indicate imputed SNPs. Dark colored points indicate previously-described sub-threshold suggestive hits, and light colored points are within 0.5Mb of the actual SNP (blue imputed, green genotyped).</p

Genome-wide significant GWAS SNPs.

<p>The genotype is coded for the risk increasing allele, which is the first one mentioned (e.g., T in T/A). The discovery test of ARHI in GERA non-Hispanic whites was two-sided^#, and all replication tests were one-sided. There are no estimates for African American cases SDS/SRT, as the sample size was too small. OR, odds ratio (for case/control phenotypes); Effect, effect size estimate (for SDS/SRT phenotypes); Freq, frequency of the risk increasing allele; SRT, speech reception threshold; SDS, speech discrimination score.</p

Characteristics of age-related hearing impairment (ARHI) cases and controls in the GERA cohort.

<p>Note that speech reception threshold (SRT) and speech discrimination score (SDS) were only available on a small portion of the cohort. For dichotomous variables, N (% of non-missing), and for continuous variables, mean (sd), unless otherwise indicated.</p

Study design.

<p>(A) GERA genotyping, (B) GERA phenotyping, (C) UK Biobank replication cohort, and (D) analysis. DX, diagnosis.</p

SNPs identified in candidate gene exon analysis.

<p>The genotype is coded for the risk increasing allele, which is the first one mentioned (e.g., A in A/C). The discovery test of ARHI in GERA non-Hispanic whites was two-sided^#, and all replication tests were one-sided. There are no estimates for African American cases SDS/SRT, as the sample size was too small. OR, odds ratio (for case/control phenotypes); Effect, effect size estimate (for SDS/SRT phenotypes); Freq, frequency of the risk increasing allele; SRT, speech reception threshold; SDS, speech discrimination score.</p

Sensorineural structures of ColI(2.3)⁺/Rs1⁺ mice are normal despite the bony overgrowth affecting the ossicular chain.

<p>(<b>A</b>) Histological analysis of the sensorineural structures of 12-week-old WT and ColI(2.3)⁺/Rs1⁺ revealed no gross abnormalities in the organ of Corti (OC), tunnel of Corti (TC), inner hair cells (IHC), or outer hair cells (OHC). (<b>B</b>) Whole mount immunofluorescence preparations of the outer hair cells revealed no visible differences in hair cell structure or number in ColI(2.3)⁺/Rs1⁺ in comparison to WT structures. (<b>C</b>) Micro computed x-ray tomography of the temporal bones was examined. A region of interest including the middle ear was selected in WT and ColI(2.3)⁺/Rs1⁺ at 12-week-old mice. ColI(2.3)⁺/Rs1⁺ temporal bones showed significant bony overgrowth of the middle ear compared to WT. The ossicles are structurally identifiable and are affected in the ColI(2.3)⁺/Rs1⁺ mice [malleus (M), incus (I), and stapes (S)].</p

Abnormal peri-lacunar remodeling in cochlear fibrous dysplasia.

<p>(<b>A</b>) 12-week-old cochlea from male WT and ColI(2.3)⁺/Rs1⁺ cochleae were examined for markers of peri-lacunar remodeling in a specific area of the otic capsule. (<b>B–D</b>) TRAP, a marker for osteoclast activity, was elevated in moderate and severe ColI(2.3)⁺/Rs1⁺ otic capsules compared to the low levels observed in WT controls. (<b>E</b>) The expression of osteocyte secreted remodeling factor, MMP-13 was significantly increased 7-fold in moderate (**, p<0.005) and 100 fold in severe ColI(2.3)⁺/Rs1⁺ cochleae (***, p<1×10⁻⁰⁵) compared to WT cochleae. (n = 6 male WT, n = 5 male ColI(2.3)⁺/Rs1⁺ moderate, and n = 5 male ColI(2.3)⁺/Rs1⁺ severe cochlea). Error bars are mean +/− SEM of triplicate measurements. (<b>F–H</b>) Immunohistochemistry showed that MMP-13 expression was confined to the otic capsule in WT cochleae, but the domain of expression is increased in ColI(2.3)⁺/Rs1⁺ moderate and severe cochlea. (<b>I–K</b>) Thionin staining of the canalicular network was examined in WT and ColI(2.3)⁺/Rs1⁺ moderate and severe cochleae. The canalicular network in WT cochlea shows normal elliptical osteocyte morphology and connectivity. ColI(2.3)⁺/Rs1⁺ moderate and severe cochleae show an abnormal rounded osteocyte morphologies with disrupted and disorganized canalicular networks of varying severity (n = 5 males per genotype).</p

Irregular lesions in ColI(2.3)⁺/Rs1⁺ cochlea involve the apex and the labyrinth causing thickening of the otic capsule.

<p>(<b>A–C</b>) Cochlea from 12-week-old WT and ColI(2.3)⁺/Rs1⁺ mice were stained for toluidine blue and examined histologically. Mixed boney fibrous lesions were often observed surrounding ColI(2.3)⁺/Rs1⁺ cochlea compared to WT controls. ColI(2.3)⁺/Rs1⁺ cochlea had multiple fibrous boney overgrowths of the vestibular bone compared to normal WT morphology. Scale bar, 150 µm. (<b>D–F</b>) The walls of ColI(2.3)⁺/Rs1⁺ cochleae showed significant thickening in comparison to WT cochleae, possibly due to the overall thickening of the surrounding otic capsule. The stria vascularis (SV) in the ColI(2.3)⁺/Rs1⁺ mice appear normal. Scale bar, 100 µm.</p