Microfluidic enrichment of plasma cells improves treatment of multiple myeloma

Abstract

Cytogenetic alterations form the basis for risk stratification for multiple myeloma (MM) and guide the selection of therapy; however, current pathology assays performed on bone marrow samples can produce false‐negatives due to the unpredictable distribution and rarity of MM cells. Here, we report on a microfluidic device used to facilitate CD45 depletion to enhance the detection of cytogenetic alterations in plasma cells (PCs). Bone marrow samples from 48 patients with MM were each divided into two aliquots. One aliquot was subjected to classic flow cytometry and fluorescent in situ hybridization (FISH). The other first went through CD45+ cell depletion, further enriched by microfluidic size selection. The enriched samples were then analyzed using flow cytometry and FISH and compared to those analyzed using the classic method only. Unlike the traditional method, the microfluidic device removed the CD45+ leukocytes and specifically selected PCs from the remaining white blood cells. Therefore, the microfluidic method (MF‐CD45‐TACs) significantly increased the percentage of CD38+/CD138+ cells to 37.7 ± 20.4% (P < 0.001) from 10.3 ± 8.5% in bone marrow. After the MF‐CD45‐TAC enrichment, the detection rate of IgH rearrangement, del(13q14), del(17p), and 1q21 gains, rose to 56.3% (P < 0.001), 37.5% (P < 0.001), 22.9% (P < 0.001), and 41.7% (P = 0.001), respectively; all rates of detection were significantly increased compared to the classically analyzed samples. In this clinical trial, this microfluidic‐assisted assay provided a precise detection of cytogenetic alterations in PCs and improved clinical outcomes.

Single-cell transcriptomes reveal the mechanism for a breast cancer prognostic gene panel

“…The inconsistency may be derived from inter-tumor and intra-tumor heterogeneity as these results were based on assays on bulky tumors. We have recently shown that a cancer relapse signaling pathway can be detected only in a single-cell transcriptome, not in a bulky tumor analysis, as bulky tumor transcriptomic analyses obscure the low signal of certain biomarkers [ 8 ]. We rationalized that single-cell transcriptomic analysis can enhance signal-to-noise ratio over bulky tumor transcriptomic analysis.…”

Hunting down the dominating subclone of cancer stem cells as a potential new therapeutic target in multiple myeloma: An artificial intelligence perspective

“…While single-cell proteomics is still uncertain, single-cell RNA-seq is a widespread practice in research laboratories now. Many microfluidic devices, including ours, have been developed for single-cell transcriptome analysis[ 17 ] but clinical application of single-cell transcriptome is still not common, especially in cancer characterization and classification. US Food and Drug Administration approved CellSearch™ (Janssen, Raritan, NJ) of circulating tumor cells (CTC) for predicting PFS and OS in metastatic breast cancer for clinical use in 2005.…”

“…We have conducted and published a small Phase I clinical trial with our devices in myeloma risk stratification of MM[ 26 ]. As a proof-of-concept for instrumentation of our prototype device, we published the clinical trial using MLSCA/MF-CD45-TACs on MM 48 patients[ 17 ], which shed new light for scale-up applications in clinics.…”

“…We applied this MLSCA for a clinical trial of risk stratification of MM[ 17 ]. As our MF-CD45-TACs differs from CellSearch™, we expected single-cell features (FACS, biomarkers) of CellSearch™ differ from those of our MF-CD45-TACs (Figure 2 ).…”

“…Other cells are forced to bypass to the next unit; B: FEOET push the droplet (with a cell) past the open valve (orange bar), which closes, locking the droplet into the 10-nl ring with RT/RT-PCR master mix (green); and C: The ring’s peristaltic pump breaks the droplet to mix the cell with master-mix. When the reaction is finished, oil (blue) pushes the product (cDNA) out of the ring in the form of a droplet (10-nL) for downstream molecular analysis (Refer to[ 17 ] for details).…”