Determination of complex subclonal structures of hematological malignancies by multiplexed genotyping of blood progenitor colonies.

Abstract

Current next generation sequencing (NGS) technologies allow unprecedented insights into the mutational profiles of tumours. Recent studies in myeloproliferative neoplasms have further demonstrated that not only the mutational profile, but also the order in which these mutations are acquired is relevant for our understanding of the disease. Our ability to assign mutation order from NGS data alone, is however limited. Here we present a strategy of highly multiplexed genotyping of burst forming unit-erythroid (BFU-E) colonies based on NGS results to assess subclonal tumour structure. This allowed for the generation of complex clonal hierarchies and determination of order of mutation acquisition far more accurately than was possible from NGS data alone.