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. 2022 Feb;40(1):57-62.
doi: 10.1002/hon.2932. Epub 2021 Oct 15.

Droplet digital polymerase chain reaction for the assessment of disease burden in hairy cell leukemia

Alessandro Broccoli  1   2 , Carolina Terragna  1 , Laura Nanni  2 , Marina Martello  1   2 , Silvia Armuzzi  1   2 , Claudio Agostinelli  2   3 , Alice Morigi  2 , Beatrice Casadei  2 , Cinzia Pellegrini  1 , Vittorio Stefoni  1   2 , Elena Sabattini  3 , Lisa Argnani  2 , Pier Luigi Zinzani  1   2
Affiliations

Droplet digital polymerase chain reaction for the assessment of disease burden in hairy cell leukemia

Alessandro Broccoli et al. Hematol Oncol. 2022 Feb.

Abstract

BRAFV600E mutation is the pathogenic driver of hairy cell leukemia (HCL) found in the vast majority of cases both at onset and during recurrences. The identification of the mutated allele in blood and marrow correlates with the presence of neoplastic cells and can be considered a marker of active disease. Likewise, the absence of the mutation after treatment may indicate a state of deep response. The BRAFV600E burden was measured by droplet digital polymerase chain reaction (ddPCR) and expressed as fractional abundance in 35 HCL patients at different stages of disease (onset, relapse, complete response [CR] after treatment, long-term remission) in peripheral blood and/or bone marrow (when available). Mean values of fractional abundance for patients at diagnosis, relapse and response, respectively, were 12.26%, 16.52% and 0.02% in peripheral blood and 23.51%, 13.96% and 0.26% in bone marrow. Four patients out of 6 evaluated at response were molecularly negative for BRAFV600E in peripheral blood. Mean fractional abundance in peripheral blood tested in 14 patients with long lasting CR was 0.05%, and 10 patients were BRAFV600E negative. These preliminary results suggest that ddPCR permits to assess the active tumor burden in HCL at different disease phases and support the hypothesis that some patients in CR qualify for a molecular CR.

Keywords: BRAF-V600E mutation; complete response; droplet digital PCR; hairy cell leukemia.

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Conflict of interest statement

The authors have stated that they have no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Positive control, BRAF V600E‐positive A375 melanoma cell‐line (panel A). Negative control, BRAF wild‐type HL60 acute myeloid leukemia cell‐line (panel B). BRAF V600E‐positive HCL patient at disease onset, peripheral blood (panel C). An HCL patient with long lasting complete response, peripheral blood (panel D)
FIGURE 2
FIGURE 2
BRAF V600E allele burden expressed as fractional abundance at each disease phase, according to sampling site. Vertical axis is in logarithmic scale. Note that the logarithmic scale does not show patients whose fractional abundance is zero
See this image and copyright information in PMC

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