Analytical validation of a novel comprehensive genomic profiling informed circulating tumor DNA monitoring assay for solid tumors.

Emerging technologies focused on the detection and quantification of circulating tumor DNA (ctDNA) in blood show extensive potential for managing patient treatment decisions, informing risk of recurrence, and predicting response to therapy. Currently available tissue-informed approaches are often limited by the need for additional sequencing of normal tissue or peripheral mononuclear cells to identify non-tumor-derived alterations while tissue-naïve approaches are often limited in sensitivity. Here we present the analytical validation for a novel ctDNA monitoring assay, FoundationOne®Tracker. The assay utilizes somatic alterations from comprehensive genomic profiling (CGP) of tumor tissue. A novel algorithm identifies monitorable alterations with a high probability of being somatic and computationally filters non-tumor-derived alterations such as germline or clonal hematopoiesis variants without the need for sequencing of additional samples. Monitorable alterations identified from tissue CGP are then quantified in blood using a multiplex polymerase chain reaction assay based on the validated SignateraTM assay. The analytical specificity of the plasma workflow is shown to be 99.6% at the sample level. Analytical sensitivity is shown to be >97.3% at ≥5 mean tumor molecules per mL of plasma (MTM/mL) when tested with the most conservative configuration using only two monitorable alterations. The assay also demonstrates high analytical accuracy when compared to liquid biopsy-based CGP as well as high qualitative (measured 100% PPA) and quantitative precision (<11.2% coefficient of variation).

Characterizing human genomic coevolution in locus-gene regulatory interactions.

BACKGROUND: Coevolution has been used to identify and predict interactions and functional relationships between proteins of many different organisms including humans. Current efforts in annotating the human genome increasingly show that non-coding DNA sequence has important functional and regulatory interactions. Furthermore, regulatory elements do not necessarily reside in close proximity of the coding region for their target genes. RESULTS: We characterize coevolution as it appears in locus-gene interactions in the human genome, focusing on expression Quantitative Trait - Locus (eQTL) interactions. Our results show that in these interactions the conservation status of the loci is predictive of the conservation status of their target genes. Furthermore, comparing the phylogenetic histories of intra-chromosomal pairs of loci and transcription start sites, we find that pairs that appear coevolved are enriched for cis-eQTL interactions. Exploring this property we found that coevolution might be useful in prioritizing association tests in cis-eQTL detection. CONCLUSIONS: The relationship between the conservation status of pairs of loci and protein coding transcription start sites reveal correlations with regulatory interactions. Pairs that appear coevolved are enriched for intra-chromosomal regulatory interactions, thus our results suggest that measures of coevolution can be useful for prediction and detection of new interactions. Measures of coevolution are genome-wide and could potentially be used to prioritize the detection of distant or inter-chromosomal interactions such as trans-eQTL interactions in the human genome.

Pluribus-Exploring the Limits of Error Correction Using a Suffix Tree.

Next generation sequencing technologies enable efficient and cost-effective genome sequencing. However, sequencing errors increase the complexity of the de novo assembly process, and reduce the quality of the assembled sequences. Many error correction techniques utilizing substring frequencies have been developed to mitigate this effect. In this paper, we present a novel and effective method called Pluribus, for correcting sequencing errors using a generalized suffix trie. Pluribus utilizes multiple manifestations of an error in the trie to accurately identify errors and suggest corrections. We show that Pluribus produces the least number of false positives across a diverse set of real sequencing datasets when compared to other methods. Furthermore, Pluribus can be used in conjunction with other contemporary error correction methods to achieve higher levels of accuracy than either tool alone. These increases in error correction accuracy are also realized in the quality of the contigs that are generated during assembly. We explore, in-depth, the behavior of Pluribus , to explain the observed improvement in accuracy and assembly performance. Pluribus is freely available at http://compbio. CASE: edu/pluribus/.