Summary of results for all specimens
| Patient (specimen) . | Days since first known MRD negative by ΔN flow cytometry . | Fusion evaluated . | ΔN flow cytometry result . | Abnormal mast cell phenotype . | RT-PCR result . | Unsorted: FISH % positive (cells) . | Sorted mast cells: FISH % positive (cells) . | Sorted T cells: FISH % positive (cells) . | Sorted myeloblasts: FISH % positive (cells) . | Sorted neutrophils: FISH % positive (cells) . | cKIT mutation status (cell population) . |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | D0 | RUNX1::RUNX1T1 | No malignancy | Yes | Positive | N/A | 64 (200) | Negative (183) | Negative (200) | N/A | N/A |
| 2 | D133 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 83.8 (37) | Negative (200) | Negative (200) | N/A | Negative (unsorted) |
| 3 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 68.4 (38) | N/A | N/A | N/A | Negative (mast cells) |
| 4 | D44 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 96 (100) | Negative (200) | Negative (200) | N/A | Low positive (unsorted) |
| 5 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 11.1 (18) | Negative (200) | Negative (200) | N/A | N/A |
| 6 | D143 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 55 (200) | Negative (199) | Negative (200) | N/A | Negative (unsorted) |
| 7 | D27 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 16.7 (6) | Negative (200) | Negative (200) | N/A | N/A |
| 8 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | Negative (200) | Negative (200) | Negative (200) | N/A | N/A |
| 9a | D0 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 3.9 (59) | Negative (200) | Negative (200) | N/A | N/A |
| 9b | D43 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 22 (100) | Negative (200) | Negative (200) | N/A | N/A |
| 10a | D66 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 27 (37) | Negative (200) | Negative (200) | N/A | N/A |
| 10b | D123 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 16 (38) | Negative (200) | Negative (200) | N/A | Negative (unsorted) |
| 11 | D24 | CBFB::MYH11 | No malignancy | No evidence | Positive | N/A | 76 (200) | N/A | Negative (200) | N/A | N/A |
| 12 | D0 | CBFB::MYH11 | No malignancy | No evidence | Positive | 11.5 (200) | 15.4 (13) | Negative (41) | Negative (19) | N/A | Low positive (unsorted) |
| 13 | D9 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | Negative (200) | Negative (200) | Negative (200) | N/A | N/A |
| 14a | D40 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 90 (200) | Negative (67) | Negative (200) | N/A | N/A |
| 14b | D173 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 93 (200) | Negative (200) | Negative (200) | N/A | N/A |
| 15 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 75.2 (101) | Negative (200) | Negative (100) | N/A | N/A |
| 16 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | Negative (100) | 95.2 (42) | N/A | N/A | N/A | N/A |
| 17 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 43.5 (200) | Negative (200) | Negative (200) | N/A | N/A |
| 18a | D164 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 15.2 (92) | Negative (202) | Negative (200) | Negative (200) | N/A |
| 18b | D204 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 63.5 (200) | Negative (200) | Negative (200) | Negative (200) | N/A |
| 18c | D385 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 68.6 (153) | Negative (200) | Negative (200) | Negative (200) | N/A |
| 18d | D413 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 78.3 (83) | Negative (80) | Negative (100) | Negative (200) | N/A |
| 19a | D79 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 93 (165) | Negative (200) | Negative (52) | N/A | Positive (mast cells) |
| 19b | D99 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 89 (200) | Negative (200) | Negative (200) | N/A | Positive (mast cells) |
| 19c | D159 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 91 (200) | Negative (200) | Negative (200) | N/A | Positive (mast cells) |
| 19d | D259 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 80 (152) | Negative (78) | Negative (159) | N/A | Positive (mast cells) |
| Patient (specimen) . | Days since first known MRD negative by ΔN flow cytometry . | Fusion evaluated . | ΔN flow cytometry result . | Abnormal mast cell phenotype . | RT-PCR result . | Unsorted: FISH % positive (cells) . | Sorted mast cells: FISH % positive (cells) . | Sorted T cells: FISH % positive (cells) . | Sorted myeloblasts: FISH % positive (cells) . | Sorted neutrophils: FISH % positive (cells) . | cKIT mutation status (cell population) . |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | D0 | RUNX1::RUNX1T1 | No malignancy | Yes | Positive | N/A | 64 (200) | Negative (183) | Negative (200) | N/A | N/A |
| 2 | D133 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 83.8 (37) | Negative (200) | Negative (200) | N/A | Negative (unsorted) |
| 3 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 68.4 (38) | N/A | N/A | N/A | Negative (mast cells) |
| 4 | D44 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 96 (100) | Negative (200) | Negative (200) | N/A | Low positive (unsorted) |
| 5 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 11.1 (18) | Negative (200) | Negative (200) | N/A | N/A |
| 6 | D143 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 55 (200) | Negative (199) | Negative (200) | N/A | Negative (unsorted) |
| 7 | D27 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | 16.7 (6) | Negative (200) | Negative (200) | N/A | N/A |
| 8 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | Low positive | N/A | Negative (200) | Negative (200) | Negative (200) | N/A | N/A |
| 9a | D0 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 3.9 (59) | Negative (200) | Negative (200) | N/A | N/A |
| 9b | D43 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 22 (100) | Negative (200) | Negative (200) | N/A | N/A |
| 10a | D66 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 27 (37) | Negative (200) | Negative (200) | N/A | N/A |
| 10b | D123 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | 16 (38) | Negative (200) | Negative (200) | N/A | Negative (unsorted) |
| 11 | D24 | CBFB::MYH11 | No malignancy | No evidence | Positive | N/A | 76 (200) | N/A | Negative (200) | N/A | N/A |
| 12 | D0 | CBFB::MYH11 | No malignancy | No evidence | Positive | 11.5 (200) | 15.4 (13) | Negative (41) | Negative (19) | N/A | Low positive (unsorted) |
| 13 | D9 | CBFB::MYH11 | No malignancy | No evidence | Low positive | N/A | Negative (200) | Negative (200) | Negative (200) | N/A | N/A |
| 14a | D40 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 90 (200) | Negative (67) | Negative (200) | N/A | N/A |
| 14b | D173 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 93 (200) | Negative (200) | Negative (200) | N/A | N/A |
| 15 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 75.2 (101) | Negative (200) | Negative (100) | N/A | N/A |
| 16 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | Negative (100) | 95.2 (42) | N/A | N/A | N/A | N/A |
| 17 | D0 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 43.5 (200) | Negative (200) | Negative (200) | N/A | N/A |
| 18a | D164 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 15.2 (92) | Negative (202) | Negative (200) | Negative (200) | N/A |
| 18b | D204 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 63.5 (200) | Negative (200) | Negative (200) | Negative (200) | N/A |
| 18c | D385 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 68.6 (153) | Negative (200) | Negative (200) | Negative (200) | N/A |
| 18d | D413 | RUNX1::RUNX1T1 | No malignancy | No evidence | N/A | N/A | 78.3 (83) | Negative (80) | Negative (100) | Negative (200) | N/A |
| 19a | D79 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 93 (165) | Negative (200) | Negative (52) | N/A | Positive (mast cells) |
| 19b | D99 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 89 (200) | Negative (200) | Negative (200) | N/A | Positive (mast cells) |
| 19c | D159 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 91 (200) | Negative (200) | Negative (200) | N/A | Positive (mast cells) |
| 19d | D259 | RUNX1::RUNX1T1 | No malignancy | Yes | N/A | N/A | 80 (152) | Negative (78) | Negative (159) | N/A | Positive (mast cells) |
Mast cells were deemed phenotypically normal or abnormal based on right angle light scatter, CD45, CD117, CD2, and CD25 expression and were evaluated by difference from normal flow cytometry (ΔN). Positive RT-PCR results indicate a normalized copy number of >0.005, and low positive results indicate a normalized copy number of 0.0005 to 0.005. cKIT mutations were evaluated for D816V in exon 17 using RT-PCR, next generation sequencing, or both; patient 12 was additionally evaluated for D816Y in exon 17 and Y418_D419insFF in exon 8. Mast cells (bright CD117+/CD34–), T cells (CD3+), myeloblasts (CD34+/CD117 heterogenous), and neutrophils (SSC/CD33+) were flow cytometrically sorted for FISH analysis. Purity of sorts was confirmed by analysis of sorted CD3+ T cells and CD33+ neutrophils when quality control was performed prior to sorting. Time point is defined as days since first known AML-MRD-negative result by ΔN flow cytometry.
N/A, not available.