Linked-read whole-genome sequencing resolves common and private structural variants in multiple myeloma

Pena Perez, L.; Frengen, N.; Hauenstein, J.; Gran, C.; Gustafsson, C.; Eisfeldt, J.; Kierczak, M.; Taborsak-Lines, F.; Olsen, R.-A.; Wallblom, A.; Krstic, A.; Ewels, P. A.; Lindstrand, A.; Mansson, R.

doi:10.1101/2021.12.09.471893

bioRxiv · 10.1101/2021.12.09.471893

Linked-read whole-genome sequencing resolves common and private structural variants in multiple myeloma

Pena Perez, L.·Frengen, N.·Hauenstein, J.·Gran, C.·Gustafsson, C.·Eisfeldt, J.·Kierczak, M.·Taborsak-Lines, F.·Olsen, R.-A.·Wallblom, A.·Krstic, A.·Ewels, P. A.·Lindstrand, A.·Mansson, R.

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Abstract

Multiple myeloma (MM) is an incurable and aggressive plasma cell malignancy characterized by a complex karyotype with multiple structural variants (SVs) and copy number variations (CNVs). Linked-read whole-genome sequencing (lrWGS) allows for refined detection and reconstruction of SVs by providing long-range genetic information from standard short-read sequencing. This makes lrWGS an attractive solution for capturing the full genomic complexity of MM. Here we show that high-quality lrWGS data can be generated from low numbers of FACS sorted cells without DNA purification. Using this protocol, we analyzed FACS sorted MM cells from 37 MM patients with lrWGS. We found high concordance between lrWGS and FISH for the detection of recurrent translocations and CNVs. Outside of the regions investigated by FISH, we identified >150 additional SVs and CNVs across the cohort. Analysis of the lrWGS data allowed for resolving the structure of diverse SVs affecting the MYC and t(11;14) loci causing the duplication of genes and gene regulatory elements. In addition, we identified private SVs causing the dysregulation of genes recurrently involved in translocations with the IGH locus and show that these can alter the molecular classification of the MM. Overall, we conclude that lrWGS allows for the detection of aberrations critical for MM prognostics and provides a feasible route for providing comprehensive genetics. Implementing lrWGS could provide more accurate clinical prognostics, facilitate genomic medicine initiatives, and greatly improve the stratification of patients included in clinical trials. KEY POINTS- Linked-read WGS can be performed without DNA purification and allows for resolving the diverse structural variants found in multiple myeloma. - Linked-read WGS can, as a stand-alone assay, provide comprehensive genetics in myeloma and other diseases with complex genomes.

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Pena Perez, L., Frengen, N., Hauenstein, J., Gran, C., Gustafsson, C., Eisfeldt, J., Kierczak, M., Taborsak-Lines, F., Olsen, R.-A., Wallblom, A., Krstic, A., Ewels, P. A., Lindstrand, A., Mansson, R.. 2021-12-09. Linked-read whole-genome sequencing resolves common and private structural variants in multiple myeloma. https://doi.org/10.1101/2021.12.09.471893

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