Adverse Prognostic Impact of the KIT D816V Transcriptional Activity in Advanced Systemic Mastocytosis
Abstract
:1. Introduction
2. Results
2.1. Patients’ Characteristics
2.2. Assessment of Analytical Sensitivity, Specificity and Reproducibility of the dPCR Assay
2.3. Inter-Laboratory Round-Robin Test
2.4. Comparison of VAF between PB and BM
2.5. Comparison between EAB and VAF
2.6. Disease Characteristics in Cohorts A and B
2.7. Prognostic Value of EAB, VAF and EAB/VAF Ratio
3. Discussion
4. Material and Methods
4.1. Patients and Samples
4.2. RNA-Based Assessment of KIT D816V
4.3. DNA-Based Assessment of KIT D816V
4.3.1. Chip-Based Digital PCR.
4.3.2. Droplet Digital PCR.
4.3.3. Quantitative Real-Time PCR.
4.3.4. Round-Robin Test for Various DNA-Based PCR Platforms
4.3.5. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
References
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Variables | ISM | AdvSM | p-Value |
---|---|---|---|
Number of patients (n) | 40 | 121 | - |
Age in years, median (range) | 54 (29–83) | 76 (30–90) | <0.0001 |
Male, n (%) | 17 (43) | 81 (67) | 0.006 |
Hemoglobin, g/dL; median (range) | 13.9 (11.7–16.8) | 10.8 (5.8–15.8) | <0.0001 |
Platelets, ×109/L; median (range) | 283 (87–461) | 114 (12–958) | 0.0002 |
MC-infiltration in BM histology, % | not applicable | 30 (0–100) | - |
Serum tryptase, µg/L; median (range) | 46 (8–166) | 180 (11–1382) | <0.0001 |
Alkaline phosphatase, U/L; median (range) | 76 (15–166) | 200 (33–1279) | <0.0001 |
Diagnosis | |||
ASM, n (%) | - | 18 (15) | - |
MCL, n (%) | - | 2 (2) | - |
SM/MCL-AHN, n (%) | - | 101 (83) | - |
Progression to | |||
Secondary AML, n (%) | - | 11 (61) | - |
Secondary MCL, n (%) | - | 7 (39) | - |
Outcome | |||
Follow-up, years, median (range) | 5 (0–21) | 3 (0–25) | n.s. |
Death, n (%) | 0 (100) | 60 (50) | <0.0001 |
Overall survival, median, years | not reached | 4.8 | <0.0001 |
Treatment | |||
Midostaurin, n (%) | 1 (3) | 57 (47) | <0.0001 |
Variables | KIT D816V EAB/VAF Ratio ≤ 2 (Cohort A) | KIT D816V EAB/VAF Ratio > 2 (Cohort B) | p-Value |
---|---|---|---|
Number of patients (n) | 77 | 44 | - |
Age in years, median (range) | 71 (30–90) | 77 (52–88) | - |
Male, n (%) | 49 (63) | 32 (73) | - |
Diagnosis | |||
ASM, n (%) | 14 (18) | 4 (11) | - |
MCL, n (%) | 2 (3) | - | - |
SM/MCL-AHN, n (%) | 61 (79) | 40 (90) | - |
AHN-subtypes | |||
MDS/MPN-u, n (%) | 18 (30) | 13 (33) | - |
CMML, n (%) | 27 (44) | 17 (43) | - |
MDS, n (%) | 5 (8) | 6 (15) | - |
MPN-eo, n (%) | 1 (2) | - | |
AML, n (%) | 1 (2) | 1 (2) | - |
CEL, n (%) | 7 (11) | 1 (2) | - |
PMF, n (%) | 2 (3) | 2 (5) | - |
Progression to | |||
AML, n (%) | 8 (10) | 3 (7) | - |
MCL, n (%) | 6 (8) | 2 (5) | - |
C-findings | |||
Hemoglobin, g/dL; median (range) | 11.4 (5.8–15.8) | 9.8 (7.5–14.5) | 0.006 |
<10 g/dL; n (%) | 20 (29) | 21 (53) | 0.01 |
Platelets, ×109/L; median (range) | 133 (12–618) | 106 (28–958) | n.s. |
<100 × 109/L, n (%) | 31 (44) | 19 (48) | n.s. |
Alkaline phosphatase, U/L; median (range) | 188 (33–1206) | 303 (53–1279) | n.s. |
>150 U/L, n (%) | 39 (57) | 31 (79) | 0.01 |
Albumin, g/L; median (range) | 38 (16–48) | 36 (22–48) | n.s. |
<34 g/L, n (%) | 23 (34) | 14 (40) | n.s. |
Ascites, n (%) | 39 (53) | 25 (61) | n.s. |
B-findings | |||
MC-infiltration in BM histology, %; median (range) | 35 (3–95) | 30 (0–100) | n.s. |
Serum tryptase, µg/L; median (range) | 170 (11–1382) | 211 (18–875) | n.s. |
>100 µg/L, n (%) | 51 (73) | 28 (74) | n.s. |
Splenomegaly, n (%) | 60 (87) | 37 (90) | n.s. |
Hepatomegaly, n (%) | 33 (52) | 28 (72) | 0.05 |
Additional SM and/or AHN relevant findings | |||
Leukocytes, ×109/L; median (range) | 10.6 (5.8–79.3) | 7.6 (1.0–89.4) | n.s. |
Monocytes, %; median (range) | 7 (1–46) | 11 (1–31) | 0.01 |
Eosinophils, %, median (range) | 3 (0–81) | 6 (0–66) | n.s. |
Vitamin B12, ng/L; median (range) | 1188 (114–6000) | 2842 (489–6000) | 0.02 |
>180 ng/L, n (%) | 50 (96) | 32 (100) | n.s. |
KIT D816V EAB in PB, %, median (range) | 30 (0–95) | 28 (2–88) | n.s. |
KIT D816V VAF in PB, %, median (range) | 27.0 (0.0–49.8) | 4.0 (0.1–30.8) | <0.001 |
GI-infiltration, n (%) | 30 (41) | 19 (43) | n.s. |
S/A/R mutation(s) a, n (%) | 38 (51) | 31 (74) | 0.02 |
Treatment | |||
Midostaurin b, n (%) | 26 (48) | 14 (39) | n.s. |
Cladribine b, n (%) | 6 (11) | 7 (19) | n.s. |
Midostaurin + cladribine b, n (%) | 22 (41) | 15 (36) | n.s. |
Response to any treatment c, n (%) | 10 (30) | 9 (45) | n.s. |
Outcome | |||
Follow-up, years, median (range) | 3.5 (0.0–24.6) | 2.2 (0.0–11.9) | - |
Death, n (%) | 33 (43) | 27 (61) | - |
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Naumann, N.; Lübke, J.; Baumann, S.; Schwaab, J.; Hoffmann, O.; Kreil, S.; Dangelo, V.; Reiter, L.; Bugert, P.; Kristensen, T.; et al. Adverse Prognostic Impact of the KIT D816V Transcriptional Activity in Advanced Systemic Mastocytosis. Int. J. Mol. Sci. 2021, 22, 2562. https://doi.org/10.3390/ijms22052562
Naumann N, Lübke J, Baumann S, Schwaab J, Hoffmann O, Kreil S, Dangelo V, Reiter L, Bugert P, Kristensen T, et al. Adverse Prognostic Impact of the KIT D816V Transcriptional Activity in Advanced Systemic Mastocytosis. International Journal of Molecular Sciences. 2021; 22(5):2562. https://doi.org/10.3390/ijms22052562
Chicago/Turabian StyleNaumann, Nicole, Johannes Lübke, Sofie Baumann, Juliana Schwaab, Oliver Hoffmann, Sebastian Kreil, Vito Dangelo, Lukas Reiter, Peter Bugert, Thomas Kristensen, and et al. 2021. "Adverse Prognostic Impact of the KIT D816V Transcriptional Activity in Advanced Systemic Mastocytosis" International Journal of Molecular Sciences 22, no. 5: 2562. https://doi.org/10.3390/ijms22052562