Identification of a Non-Coding Causative Variant Underlying Warsaw Breakage Syndrome Using Long-Read Based Genomic Sequencing and Transcriptome Analysis.

Currently, exome and genome sequencing achieve a diagnostic rate of 30%-50% for rare genetic diseases. With multi-modal technologies profiling the genome, transcriptome, and epigenome, interrogation of genomic elements outside of protein-coding regions shows potential to improve this as demonstrated herein. Siblings with sensorineural hearing loss, microcephaly, intellectual impairment, and growth restriction were seen in consultation. Following extensive clinical testing, long-read whole genome and cDNA-based transcriptome sequencing on the Oxford Nanopore platform identified a homozygous 1.6 kb deletion of the 5' UTR and promoter region of DDX11, a gene associated with Warsaw breakage syndrome. The deletion included the hypomethylated CpG island regulating DDX11, led to a loss of expression of DDX11 mRNA and protein, and resulted in the characteristic "railroad chromosome." Identifying a causal variant for this family required expanding the search space for genomic variants beyond protein-coding regions, and multi-modal data integration enabled a more holistic approach to variant prioritization and classification prior to pursuing targeted protein and functional assays. This multi-modal genome-wide approach heralds promise for patients on the diagnostic odyssey and who have exhausted standard of care testing.