Background Whole exome sequencing (WES) is a powerful technique for Mendelian disease gene discovery. However, variant prioritization remains a challenge. We applied WES to identify the causal variant in a large family with familial dilated cardiomyopathy (DMC) of unknown etiology. Methods and Results A large family with autosomal dominant, familial DCM was identified. Exome capture and sequencing was performed in 3 remotely related, affected subjects predicted to share <0.1% of their genomes by descent. Shared variants were filtered for rarity, evolutionary conservation, and predicted functional significance, and remaining variants were filtered against 71 locally generated exomes. Variants were also prioritized using the Variant Annotation Analysis and Search Tool (VAAST). Final candidates were validated by Sanger sequencing and tested for segregation. There were 664 shared heterozygous nonsense, missense, or splice site variants, of which 26 were rare (minor allele frequency ≤ 0.001 or not reported) in two public databases. Filtering against internal exomes reduced the number of candidates to 2, and of these, a single variant (c.1907 G>A) in RBM20, segregated with disease status and was absent in unaffected internal reference exomes. Bioinformatic prioritization with VAAST supported this result. Conclusions WES of remotely related DCM subjects from a large, multiplex family, followed by systematic filtering, identified a causal RBM20 mutation without the need for linkage analysis.

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Idiopathic dilated cardiomyopathy (DCM) is a primary myocardial disorder characterized by ventricular chamber enlargement and systolic dysfunction. Twenty to fifty percent of idiopathic DCM cases are thought to have a genetic cause. Of more than 30 genes known to be associated with DCM, rare variants in the VCL and MYBPC3 genes have been reported in several cases of DCM. In this report, we describe a family with DCM and congenital abnormalities who carry a novel missense mutation in the VCL gene. More severely affected family members also possess a second missense variant in MYBPC3, raising the possibility that this variant may be a disease modifier. Interestingly, many of the affected individuals also have congenital defects, including two with bicuspid aortic valve with aortic regurgitation. We discuss the implications of the family history and genetic information on management of at-risk individuals with aortic regurgitation.

Background: The purpose of this study was to investigate the association between haplotype-tagging single nucleotide polymorphisms (SNPs) within the Aurora Kinase A (AURKA) gene and prostate cancer outcomes in patients with clinically localized prostate cancer. Methods:Four intronic haplotype-tagging SNPs within the AURKA gene were individually selected and examined in regard to their influence on clinical outcomes in 212 patients who underwent radical prostatectomy as first-line treatment. Haplotype-tagging SNPs were selected using the ABI SNP Browser to cover SNPs with an r 2 of 0.90 or greater in the AURKA gene with a minor allele frequency of at least 0.25. Results:In our study, a log-rank univariate analysis was performed to identify significant associations between probability of recurrence-free survival or disease-free survival and known prognostic indicators as well as AURKA genotypes. The prognostic indicators Gleason grade, surgical margin, extracapsular extension, and disease stage were associated with recurrence-free survival (all p<0.001). Only Gleason grade was associated with disease-free survival (p<0.001). No associations were found for the SNPs rs1468055, rs8117896, rs2064863, and rs1468056 analyzed for either RFS (p=0.7213, p=0.5140, p=0.0714, p=0.4731, respectively) or DFS (p=0.3282, p=0.1909, p=0.4111, p=0.5014, respectively). Clinical data on patient follow-up at Vanderbilt University Medical Center were retrospectively gathered via electronic medical records. The mean follow-up for disease-free survival and assessment of prostate cancer progression were 8.3 ± 2.4-y and 4.4 ± 3.9-y, respectively. The endpoints analyzed were recurrence-free survival (RFS) and diseasefree survival (DFS). Recurrence post-prostatectomy was classified as biochemical (which has been shown to be a poor predictor of overall survival in patients with localized prostate cancer treated with radical prostatectomy) [14], local or distant, and the most advanced recurrence type documented was assigned to each patient. Biochemical recurrence was defined as a prostate-specific antigen (PSA) detection of > 0.1 ng/ ml in at least two consecutive tests. DFS was defined as the length of time between the date of prostatectomy to the date of death or last follow-up. Conclusions Genotyping of Aurora Kinase A polymorphisms in prostate cancer samplesProstatectomy specimens were processed and genomic DNA extracted from deparaffinizedspecimens as described previously [15]. Purified genomic DNA was genotyped for the following haplotypetagging SNPs in the AURKA gene: rs1468055, rs8117896, rs2064863, and rs1468056. Haplotype-tagging SNPs (htSNPs) were selected using the ABI SNP Browser to cover SNPs with an r 2 of 0.90 or greater in the AURKA gene with a minor allele frequency (MAF) of at least 0.25. A total of 4 SNPs were selected and genotyped in our patient cohort. Allelic discrimination of these AURKA polymorphisms was performed using Taqman SNP genotyping assays (Applied Biosystems: and C_8947664_10 [rs1468056]). The fina...

Background53BP1 binds to the tumor suppressor p53 and has a key role in DNA damage response and repair. Low 53BP1 expression has been associated with decreased survival in breast cancer and has been shown to interact with several prognostic factors in non-small cell lung cancer. The role of 53BP1 in pancreatic ductal adenocarcinoma (PDAC) has yet to be determined. We aimed to investigate whether 53BP1 levels interact with established prognostic factors in PDAC.Methods106 patients for whom there was tissue available at time of surgical resection for PDAC were included. A tissue microarray was constructed using surgical specimens, stained with antibodies to 53BP1, and scored for expression intensity. Univariate and multivariate statistical analyses were performed to investigate the association between 53BP1 and patient survival with known prognostic factors for survival.ResultsThe association of 53BP1 with several established prognostic factors was examined, including stage, tumor grade, surgical margin, peripancreatic extension, lymph node ratio (LNR), and CA 19–9. We found that 53BP1 modified the effects of known prognostic variables including LNR and CA 19–9 on survival outcomes. When 53BP1 intensity was low, increased LNR was associated with decreased OS (HR 4.84, 95% CI (2.26, 10.37), p<0.001) and high CA19-9 was associated with decreased OS (HR 1.72, 95% CI (1.18, 2.51), p=0.005). When 53BP1 intensity was high, LNR and CA19-9 were no longer associated with OS (p=0.958 and p=0.606, respectively).ConclusionsIn this study, 53BP1, a key player in DNA damage response and repair, was found to modify the prognostic value of two established prognostic factors, LNR and CA 19–9, suggesting 53BP1 may alter tumor behavior and ultimately impact how we interpret the value of other prognostic factors.