Supplementary Tables
NK1
NK2
NK3
NK4
NK5
NK6
NK7
NK8
NK9
NK10
NK11
NK12
NK13
NK14
NK15
NK16
NK17
NK18
NK19
NK20
NK21
NK22
EZH2
TP53
CEBPα
RUNX1
DNMT3A
KIT
kRAS
nRAS
cCBL
BRAF
JAK2
MPL
IDH2
IDH1
ASXL1
NPM
AML
Patient
FLT3
Supplementary Table I. Mutational screening of primary NK-AML cohort. A total of 96 amplicons were
sequenced using Roche 454 parallel sequencing for mutational ‘hotspots’ in NPM1 exon 11, FLT3-ITD/TKD,
ASXL1 – exon 12, IDH1 – exon 4 (R132), IDH2 – exon 4 (R140) and 5 (R172), MPL – exon 10, JAK2 – exon 12 and
14, BRAF – exon 11 and 15, cCBL – exons 7, 8 and 9, nRAS – exon 2 and 3, kRAS – exon 2 and 3 and KIT – exon
17. The complete coding regions were sequenced for DNMT3a, RUNX1, CEBPα, TP53 and EZH2.
Chromosome
Instability
13q UPD
UPD 6p, 11p
13q UPD
8p UPD
13q UPD
8p UPD
6p UPD
13q UPD
Gain 1q
Gain 5q
-
Supplementary Table II. Summary of HR mediated events in primary AML. NK – primary normal
karyotype AML
Cell
And treatment
Targeted HR
(Mean % cells)
DR1/DR2
DR1/DR2 +
Spontaneous
I-SCEI HR
HR
RAD51 foci %
SCE (per
metaphase)
NK1(FLT3-ITD)
NK1(FLT3-ITD) +
AC220
0.1
0.9
17  1
12  1
0
0.3
12  2
8 0.5
NK4(FLT3-ITD)
NK4(FLT3-ITD) +
AC220
NK4(FLT3-ITD) +
NAC
0.2
1.1
15  1
10.5  0.5
0
0.1
11  2
7  0.5
0
0.2
10  2
6  0.5
NK10(FLT3-ITD)
NK10(FLT3-ITD)
+ AC220
0.1
1.8
16  0.5
9  0.5
0
0.05
11  2
7  0.5
NK14(FLT3-ITD)
NK14(FLT3-ITD)
+ AC220
0.1
0.9
13  2
12  1
0
0.05
10  1
8  0.5
NK17(WT FLT3)
NK17(WT FLT3)
+ AC220
0
0.3
81
7  0.5
0
0.1
91
7  0.5
NK20(WT FLT3)
NK20(WT FLT3)
+ AC220
0
0.2
51
6  0.5
0
0.1
42
51
Supplementary Table III. Summary of HR events in mutant FLT3-ITD and WT FLT3 cells
Cell
MOLM-13
(FLT3-ITD)
MOLM-13 +
AC220
MOLM-13 +
NAC
Targeted Recombination
(Mean %  SEM GFP cells)
DR1/DR2
DR1/DR2 +
Spontaneous HR
I-SCEI HR
0.3  0.1
2.4  0.4
RAD51 foci % (
SEM)
SCE  SEM (per
metaphase)
21  3
9  0.5
0
0.4  0.2
12  2.5
6  0.5
0
0.3  0.2
15  1
7  0.5
U937
(WT FLT3)
U937 +
AC220
U937 + NAC
0
0.3  0.1
12  2.9
5 1
0
0.1  0.1
12  1.5
5 1
0
0.2  0.1
11  1
5 1
U937 EV
U937 FLT3
U937 FLT3-ITD
U937 FLT3-ITD
+
AC220
U937 + FLT3ITD + NAC
0.1
0.1  0.1
0.4  0.1
0.7  0.3
0.9  0.5
2.8  0.4
12  2.5
17  2.5
26  1
4.5  0.5
6.5  0.5
10  1
0.2  0.1
0.3  0.2
14  2.5
5  0.5
0.3  0.1
0.2  0.2
17  2
6  0.5
Supplementary Table IV. Spontaneous mutant rate and frequency of TK deficient mutants. SEM results from
three separate experiments.
Spontaneo
us.
Mutants
Mutation
Rate
Mutation
Frequency
Empty
Vector
(FUGW)
WT-FLT3
FLT3-ITD
FLT3-ITD
FLT3-ITD
Empty
Vector
(FUGW)
(FUGW)
(FUGW)
(FUGW)
+
+
+
25nM
AC220
5mM NAC
25nM
AC220
(pMSCV)
WT FLT3
FLT3-ITD
FLT3-ITD
(pMSCV)
(pMSCV)
(pMSCV)
Cisplatin
1.5 X 10-6
1.9 X 10-5
4.3 X 10-5
8.9 X 10-5
3.2 X 10-5
3.7 X 10-5
3.6 X 10-5
3.3 X 10-5
7.9 X 10-5
4.9 X 10-5
8.3 X 10-4
0.8X10-6
1.8X 10-6
2.7X10-6
1.8X10-6
2.2X10-6
4.2X10-6
2.7X10-6
1.7X10-6
2.6X10-6
2.6X10-6
2.7X10-6
4.3 X 10-6
1.1 X 10-5
3.0 X 10-5
6.1 X 10-5
3.6 X 10-5
4.2 X 10-5
2.1 X 10-5
3.3 X 10-5
8.5 X 10-5
5.9 X 10-5
1.5 X 10-4
1.8X 10-6
0.6X 10-6
2.1X 10-6
3.8X 10-6
2.8X 10-6
7.1X 10-6
2.5X 10-6
2.7X 10-6
5.7X 10-6
1.6X10-6
3.6X 10-6