A cost-effective quasi single-cell assay for deciphering of clonal architecture of leukemic cells

Introduction Clonality assessment using IG/TCR rearrangements is an essential tool for clonal evolution analysis and minimal residual disease monitoring in acute lymphoblastic leukemia (ALL). Here we present a new multiplex method for ALL clonal structures analysis at the single-cell level. Material and Methods: DNA from bone marrow (BM) samples of 20 T-ALL and 60 B-ALL patients was used for initial rearrangements detection by targeted high-throughput sequencing. The nuclei from all 80 BM samples were extracted, measured, pooled together evenly and sorted by FACS in 96-tube plate 75 nuclei per tube. The detection of initially identified IG/TCR was performed in all nuclei aliquots. Results: The presence of all patient-specific IG/TCR was detected and analyzed for each sample. The clonal structure was resolved by pairing of clonal and subclonal rearrangements in aliquots. Discussion: The developed method is a useful tool for single-cell level clonality analysis and can be easily implemented in routine ALL diagnostics. The work is supported by RSF grant 18-14-00244, RFBR grants 20-015-00462 and 18-29-09132 and Charity foundation Podari Zhizn.

Publication History

Article published online:
13 May 2020