DNA Damage and DNA Damage Response in Chronic Myeloid Leukemia
Abstract
:1. Introduction
2. Results
2.1. γH2AX Foci in PBMCs of Healthy Donors and CML Patients
2.2. Co-Localization of γH2AX and 53BP1 Foci
2.3. DNA Damage Response
3. Discussion
4. Materials and Methods
4.1. Blood Samples
4.2. Cytology, Cytogenetics, and Molecular Analyses
4.3. Immunofluorescence Staining of γH2AX and 53BP1
4.4. Western Blotting
4.5. Statistics
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AP | Accelerated phase |
ACA | Additional chromosomal aberrations |
BP | Blast phase |
CML | Chronic myeloid leukemia |
CP | Chronic phase |
DASA | Dasatinib |
DDR | DNA damage response |
DMR | Deep molecular response |
DSB | DNA double-strand breaks |
IMA | Imatinib |
MMEJ | Microhomology mediated end-joining |
MMR | Major molecular response |
MR | Molecular response |
NHEJ | Non-homologous end joining |
NILO | Nilotinib |
PBMC | Peripheral blood mononuclear cells |
PONA | Ponatinib |
ROS | Reactive oxygen species |
TKI | Tyrosine kinase inhibitor |
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Gp | Pt | Age/Sex | Disease | TKI | γH2AX Foci Per Nucleus ± SEM | BCR/ABL MR | BCR/ABL Transcript | Cytogenetics/FISH |
---|---|---|---|---|---|---|---|---|
Group 1 | HEALTHY#1 | 49/♂ | Healthy | - | 0.6 ± 0.1 | - | - | - |
HEALTHY#2 | 62/♀ | Healthy | - | 0.7 ± 0.1 | - | - | - | |
HEALTHY#3 | 43/♂ | Healthy | - | 0.7 ± 0.1 | - | - | - | |
HEALTHY#4 | NA | Healthy | - | 0.9 ± 0.2 | - | - | - | |
HEALTHY#5 | 54/♀ | Healthy | - | 1.2 ± 0.2 | - | - | - | |
HEALTHY#6 | 36/♀ | Healthy | - | 1.2 ± 0.2 | - | - | - | |
HEALTHY#7 | 54/♀ | Healthy | - | 1.3 ± 0.1 | - | - | - | |
HEALTHY#8 | 53/♀ | Healthy | - | 1.6 ± 0.2 | - | - | - | |
Group 2 | CP-CML DMR#1 | 61/♀ | CP-CML | DASA | 0.1 ± 0.1 | MR4.5 | e13a2 (b2a2) | 46,XX,t(9;22)(q34;q11) [25] |
CP-CML DMR#2 | 64/♀ | CP-CML | Stop | 0.4 ± 0.1 | MR4.5 | e13a2 (b2a2) | NA | |
CP-CML DMR#3 | 74/♀ | CP-CML | Stop | 0.7 ± 0.2 | MR4.5 | e13a2/e14a2 (b2a2/b3a2) | NA | |
CP-CML DMR#4 | 48/♂ | CP-CML | Stop | 0.7 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#5 | 63/♀ | CP-CML | Stop | 0.8 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#6 | 55/♂ | CP-CML | DASA | 0.8 ± 0.2 | MR4 | e14a2 (b3a2) | NA | |
CP-CML DMR#7 | 61/♀ | CP-CML | Stop | 0.8 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#8 | 33/♀ | CP-CML | DASA | 0.8 ± 0.2 | MR4 | e13a2 (b2a2) | NA | |
CP-CML DMR#9 | 58/♂ | CP-CML | Stop | 0.8 ± 0.2 | MR5 | e14a2 (b3a2) | NA | |
CP-CML DMR#10 | 72/♂ | CP-CML | Stop | 0.9 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#11 | 34/♂ | CP-CML | DASA | 0.9 ± 0.1 | MR4 | e13a2 (b2a2) | NA | |
CP-CML DMR#12 | 68/♀ | CP-CML | DASA | 0.9 ± 0.2 | MR4 | e13a2 (b2a2) | NA | |
CP-CML DMR#13 | 78/♀ | CP-CML | NILO | 1.0 ± 0.2 | MR4.5 | e13a2/e14a2 (b2a2/b3a2) | NA | |
CP-CML DMR#14 | 59/♀ | CP-CML | IMA | 1.0 ± 0.2 | MR4 | e13a2 (b2a2) | NA | |
CP-CML DMR#15 | 60/♀ | CP-CML | DASA | 1.1 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#16 | 67/♀ | CP-CML | Stop | 1.1 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#17 | 76/♀ | CP-CML | Stop | 1.1 ± 0.2 | MR4.5 | e13a2 (b2a2) | NA | |
CP-CML DMR#18 | 77/♂ | CP-CML | Stop | 1.2 ± 0.2 | MR4.5 | e13a2/e14a2 (b2a2/b3a2) | NA | |
CP-CML DMR#19 | 84/♀ | CP-CML | IMA | 1.3 ± 0.2 | MR4 | e13a2 (b2a2) | NA | |
CP-CML DMR#20 | 71/♀ | CP-CML | Stop | 1.3 ± 0.3 | MR5 | e14a2 (b3a2) | NA | |
CP-CML DMR#21 | 63/♂ | CP-CML | Stop | 1.3 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML DMR#22 | 62/♂ | CP-CML | DASA | 1.7 ± 0.2 | MR4.5 | e14a2 (b3a2) | NA | |
CP-CML MMR#1 | 70/♀ | CP-CML | IMA | 0.8 ± 0.2 | 0.04% | e13a2 (b2a2) | NA | |
CP-CML MMR#2 | 59/♀ | CP-CML | Stop | 1.2 ± 0.2 | 0.07% | e13a2/e14a2 (b2a2/b3a2) | NA | |
CP-CML MMR#3 | 77/♀ | CP-CML | DASA | 1.2 ± 0.2 | 0.09% | e14a2 (b3a2) | NA | |
CP-CML MMR#4 | 54/♂ | CP-CML | DASA | 1.3 ± 0.2 | 0.02% | e14a2 (b3a2) | NA | |
Group 3 | CP-CML loss MMR#1 | 54/♀ | CP-CML | NILO | 1.0 ± 0.2 | 0.18% | e1a2 | 46,XX,t(9;22)(q34;q11) [25] |
CP-CML loss MMR#2 | 46/♂ | CP-CML | DASA | 1.1 ± 0.2 | 45% | e14a2 (b3a2) | 51,XY,+6,+8,+8,+8, t(9;22)(q34;q11),+19 [25] | |
CP-CML loss MMR#3 | 81/♂ | CP-CML | Stop | 1.3 ± 0.2 | 0.35% | e13a2/e14a2 (b2a2/b3a2) | NA | |
CP-CML loss MMR#4 * | 83/♀ | CP-CML | Stop | 2.4 ± 0.2 | 1.00% | e14a2 (b3a2) | NA | |
CP-CML loss MMR#5 | 83/♀ | CP-CML | Stop | 3.0 ± 0.2 | 30% | e13a2/e14a2 (b2a2/b3a2) | NA | |
Group 4 | CP-CML de novo#1 | 19/♂ | CP-CML | - | 1.4 ± 0.2 | 60% | e13a2 (b2a2) | 46,XY,t(9;22)(q34;q11) [25] |
CP-CML de novo#2 | 79/♀ | CP-CML | - | 1.6 ± 0.3 | 63% | e13a2 (b2a2) | 46,XX,t(9;22)(q34;q11) [25] | |
CP-CML de novo#3 | 18/♀ | CP-CML | - | 2.3 ± 0.3 | 68% | e13a2 (b2a2) | 46,XX,t(9;22)(q34;q11) [25] | |
CP-CML de novo#4 | 66/♀ | CP-CML | - | 3.4 ± 0.2 | 29% | e14a2 (b3a2) | 46,XX,t(9;22)(q34;q11) [25] | |
CP-CML de novo#5 | 53/♀ | CP-CML | - | 3.7 ± 0.3 | 68% | e13a2 (b2a2) | 46,XX,t(9;22)(q34;q11) [25] | |
Group 5 | BP-CML#1 ** | 76/♀ | BP-CML | Stop | 3.4 ± 0.4 | 32% | e13a2 (b2a2) | NA (rejected) |
BP-CML#2 | 63/♂ | BP-CML | DASA | 4.0 ± 0.4 | 63% | e14a2 (b3a2) | 46,XY,inv(3)(q21;q26), t(9;22)(q34;q11) [25] | |
BP-CML#3 *** | 38/♀ | BP-CML | PONA | 5.8 ± 0.4 | 256% | e13a2 (b2a2) | 46,XX,t(9;22)(q34;q11) [1] 44,XX,der(3)t(3;9)(p11;q11) t(9;22)(q34;q11),−7,−9, der(13)t(7;13)(q22;q34),der(22) t(9;22)(q34;q11) [5] 46,XX [14] |
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Popp, H.D.; Kohl, V.; Naumann, N.; Flach, J.; Brendel, S.; Kleiner, H.; Weiss, C.; Seifarth, W.; Saussele, S.; Hofmann, W.-K.; et al. DNA Damage and DNA Damage Response in Chronic Myeloid Leukemia. Int. J. Mol. Sci. 2020, 21, 1177. https://doi.org/10.3390/ijms21041177
Popp HD, Kohl V, Naumann N, Flach J, Brendel S, Kleiner H, Weiss C, Seifarth W, Saussele S, Hofmann W-K, et al. DNA Damage and DNA Damage Response in Chronic Myeloid Leukemia. International Journal of Molecular Sciences. 2020; 21(4):1177. https://doi.org/10.3390/ijms21041177
Chicago/Turabian StylePopp, Henning D., Vanessa Kohl, Nicole Naumann, Johanna Flach, Susanne Brendel, Helga Kleiner, Christel Weiss, Wolfgang Seifarth, Susanne Saussele, Wolf-Karsten Hofmann, and et al. 2020. "DNA Damage and DNA Damage Response in Chronic Myeloid Leukemia" International Journal of Molecular Sciences 21, no. 4: 1177. https://doi.org/10.3390/ijms21041177