Imaging flow cytometry shows monosomy 17 in circulating plasma cells in myeloma

Lam, Stephanie; Mincherton, Thomas; Hui, Henry; Sidiqi, M Hasib; Fuller, Kathryn A.; Erber, Wendy N.

doi:10.1016/j.pathol.2021.12.296

Cited by 5 publications

(6 citation statements)

References 12 publications

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“…This exquisite sensitivity was achievable because of the combination of antigenic cell identification and number of acquired cells. This low limit of detection complements the findings of Lam et al who reported detection of monosomy 17 in CD38/CD138-positive plasma cells in the blood and the marrow when only being present in small numbers (0.0039% and 0.15% of nucleated cells, respectively) 45…”

Section: Discussionsupporting

confidence: 87%

IGHcytogenetic abnormalities can be detected in multiple myeloma by imaging flow cytometry

et al. 2022

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AimsCytogenetic abnormalities involving the IGH gene are seen in up to 55% of patients with multiple myeloma. Current testing is performed manually by fluorescence in situ hybridisation (FISH) on purified plasma cells. We aimed to assess whether an automated imaging flow cytometric method that uses immunophenotypic cell identification, and does not require cell isolation, can identify IGH abnormalities.MethodsAspirated bone marrow from 10 patients with multiple myeloma were studied. Plasma cells were identified by CD38 and CD138 coexpression and assessed with FISH probes for numerical or structural abnormalities of IGH. Thousands of cells were acquired on an imaging flow cytometer and numerical data and digital images were analysed.ResultsUp to 30 000 cells were acquired and IGH chromosomal abnormalities were detected in 5 of the 10 marrow samples. FISH signal patterns seen included fused IGH signals for IGH/FGFR3 and IGH/MYEOV, indicating t(4;14) and t(11;14), respectively. In addition, three IGH signals were identified, indicating trisomy 14 or translocation with an alternate chromosome. The lowest limit of detection of an IGH abnormality was in 0.05% of all cells.ConclusionsThis automated high-throughput immuno-flowFISH method was able to identify translocations and trisomy involving the IGH gene in plasma cells in multiple myeloma. Thousands of cells were analysed and without prior cell isolation. The inclusion of positive plasma cell identification based on immunophenotype led to a lowest detection level of 0.05% marrow cells. This imaging flow cytometric FISH method offers the prospect of increased precision of detection of critical genetic lesions involving IGH and other chromosomal defects in multiple myeloma.

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Section: Discussionsupporting

confidence: 87%

IGHcytogenetic abnormalities can be detected in multiple myeloma by imaging flow cytometry

et al. 2022

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“…This was achieved as many thousands of cells were analyzed, and FISH signals were assessed in gated cells with the phenotype of interest (i.e., CD38/CD138‐positive) ensuring plasma cell specificity. This adds to the literature of imaging flow cytometry which had previously shown to be able to identify IgH rearrangements and hyperdiploidy in myeloma 14–16 …”

Section: Discussionmentioning

confidence: 80%

“…This adds to the literature of imaging flow cytometry which had previously shown to be able to identify IgH rearrangements and hyperdiploidy in myeloma. [14][15][16] Most FISH methods require positive isolation or enrichment of plasma cells to increase the yield of cells of interest prior to chromosomal analysis. In the present study, the only prior sample handling was red cell lysis or density gradient separation to remove erythrocytes and granulocytic cells.…”

Section: Discussionmentioning

confidence: 99%

“…11,12 A relatively new imaging flow cytometric method has been shown to be able to assess chromosomes by FISH in specific cell populations based on their immunophenotype, initially on chronic lymphocytic leukemia. [13][14][15] This technique, called 'immuno-flowFISH' combines immunophenotyping and FISH with analysis on an imaging flow cytometer and has been shown to be able to identify numerical (e.g., hyperdiploidy) and structural chromosomal abnormalities (e.g., IGH translocations) in antigen-positive cells in hematological malignancies. 14,16,17 In the present study, we aimed to determine the potential of immuno-flowFISH to identify abnormalities of chromosome 17, and specifically del(17p), in phenotypically identified plasma cells in both bone marrow and blood of patients with myeloma.…”

Section: Introductionmentioning

confidence: 99%

“…[13][14][15] This technique, called 'immuno-flowFISH' combines immunophenotyping and FISH with analysis on an imaging flow cytometer and has been shown to be able to identify numerical (e.g., hyperdiploidy) and structural chromosomal abnormalities (e.g., IGH translocations) in antigen-positive cells in hematological malignancies. 14,16,17 In the present study, we aimed to determine the potential of immuno-flowFISH to identify abnormalities of chromosome 17, and specifically del(17p), in phenotypically identified plasma cells in both bone marrow and blood of patients with myeloma. We hypothesized that this approach would give greater specificity than standard FISH and be able to assess del(17p) with lower-level plasma cell burden.…”

Section: Introductionmentioning

confidence: 99%

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Imaging flow cytometric detection of del(17p) in bone marrow and circulating plasma cells in multiple myeloma

Mincherton,

Lam,

Clarke

et al. 2024

Int J Lab Hematology

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BackgroundDetection of del(17p) in myeloma is generally performed by fluorescence in situ hybridization (FISH) on a slide with analysis of up to 200 nuclei. The small cell sample analyzed makes this a low precision test. We report the utility of an automated FISH method, called “immuno‐flowFISH”, to detect plasma cells with adverse prognostic risk del(17p) in bone marrow and blood samples of patients with myeloma.MethodsBone marrow (n = 31) and blood (n = 19) samples from 35 patients with myeloma were analyzed using immuno‐flowFISH. Plasma cells were identified by CD38/CD138‐immunophenotypic gating and assessed for the 17p locus and centromere of chromosome 17. Cells were acquired on an AMNIS ImageStreamX MkII imaging flow cytometer using INSPIRE software.ResultsChromosome 17 abnormalities were identified in CD38/CD138‐positive cells in bone marrow (6/31) and blood (4/19) samples when the percent plasma cell burden ranged from 0.03% to 100% of cells. Abnormalities could be identified in 14.5%–100% of plasma cells.ConclusionsThe “immuno‐flowFISH” imaging flow cytometric method could detect del(17p) in plasma cells in both bone marrow and blood samples of myeloma patients. This method was also able to detect gains and losses of chromosome 17, which are also of prognostic significance. The lowest levels of 0.009% (bone marrow) and 0.001% (blood) for chromosome 17 abnormalities was below the detection limit of current FISH method. This method offers potential as a new means of identifying these prognostically important chromosomal defects, even when only rare cells are present and for serial disease monitoring.

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