Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 23, Issue 24, Pages -Publisher
MDPI
DOI: 10.3390/ijms232415691
Keywords
multiple myeloma; single-cell RNA sequencing; clonal biomarker; V(D)J rearrangement
Funding
- European Research Council under the European Union [817997]
- Associazione Italiana Ricerca sul Cancro [IG25739]
- Umberto Veronesi Foundation
- Pfizer Global Medical Grants [75340503]
- Italian Ministry of Health-Current research IRCCS
- European Research Council (ERC) [817997] Funding Source: European Research Council (ERC)
Ask authors/readers for more resources
Multiple myeloma (MM) has a heterogeneous genetic background, making it difficult to track. However, each MM patient's malignant plasma cells share unique V(D)J rearranged sequences, which can serve as ideal disease biomarkers. By using single-cell RNA sequencing, the dominant clonotype in each sample can be easily identified, and clonal productive rearrangements can be accurately detected. This method may be used to track rare clonal cells and lay the foundation for functional single-cell analysis of minimal residual disease.
Multiple myeloma (MM) has a highly heterogeneous genetic background, which complicates its molecular tracking over time. Nevertheless, each MM patient's malignant plasma cells (PCs) share unique V(D)J rearranged sequences at immunoglobulin loci, which represent ideal disease biomarkers. Because the tumor-specific V(D)J sequence is highly expressed in bulk RNA in MM patients, we wondered whether it can be identified by single-cell RNA sequencing (scRNA-seq). To this end we analyzed CD138(+) cells purified from bone marrow aspirates of 19 samples with PC dyscrasias by both a standard method based on bulk DNA and by an implementation of the standard 10x Genomics protocol to detect expressed V(D)J sequences. A dominant clonotype was easily identified in each sample, accounting on average for 83.65% of V(D)J-rearranged cells. Compared with standard methods, scRNA-seq analysis proved highly concordant and even more effective in identifying clonal productive rearrangements, by-passing limitations related to the misannealing of consensus primers in hypermutated regions. We next validated its accuracy to track 5 clonal cells with absolute sensitivity in a virtual sample containing 3180 polyclonal cells. This shows that single-cell V(D)J analysis may be used to find rare clonal cells, laying the foundations for functional single-cell dissection of minimal residual disease.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available