Table S1: Comparison of Pretreatment Characteristics of 509 AML Patients

Thol et al. MSI2 and NUMB expression in AML– supplemental data

Supplemental Tables and Figures

Table S1. Comparison of pretreatment characteristics of CN-AML patients according to MSI2 expression levels.

Characteristic / High MSI2 expressers / Low MSI2 expressers / P
(n=74) / (n=163)
Age, years / .88
Median / 47.5 / 48
range / 25-60 / 17-60
Sex / .13
male - no. (%) / 34 (46) / 92 (56)
female - no. (%) / 40 (54) / 71 (44)
FAB-Subtype / .48
M0 - no. (%) / 2 (3) / 5 (3)
M1 - no. (%) / 22 (29) / 24 (15)
M2 - no. (%) / 14 (19) / 38 (23)
M4 - no. (%) / 28 (38) / 55 (34)
M5 - no. (%) / 3 (4) / 31 (19)
M6 - no. (%) / 2 (3) / 4 (2)
M7 - no. (%) / 2 (3) / 1 (1)
missing data - no. (%) / 1 (1) / 5 (3)
Peripheral blood blasts / .22
median (%) / 68.5 / 46
missing data – no. (%) / 6 (8) / 5 (3)
Bone marrow blasts / .8
median (%) / 80 / 80
missing data – no. (%) / 7 (9) / 12 (7)
Type of AML / .079
de novo - no. (%) / 71 (96) / 145 (89)
secondary - no. (%) / 3 (4) / 18 (11)
WBC count / .42
median - (x109/l) / 37.4 / 22.81
range - (x109/l) / 0.65-239.4 / 0.5-229.7
missing data – no. (%) / (0) / (0)
Hemoglobin / .56
median – g/L / 9.2 / 8.95
range – g/L / 5.5-12.9 / 3-15.4
missing data – no. (%) / 3 (4) / 5 (3)
Platelet count / .63
median - (x109/l) / 45 / 53
range - (x109/l) / 9-336 / 4-483
missing data – no. (%) / 3 (4) / 6 (4)
ECOG performance status / .024
0 - no. (%) / 9 (12) / 42 (26)
1 - no. (%) / 53 (72) / 104 (64)
2 - no. (%) / 12 (16) / 14 (8)
missing data - no. (%) / 0 (0) / 3 (2)
FLT3-ITD – no. (%) / <.001
mutated– no. (%) / 42 (57) / 37 (23)
Wildtype - no. (%) / 31 (42) / 122 (75)
missing – no. (%) / 1 (1) / 4 (2)
NPM1 / <.001
mutated – no. (%) / 59 (80) / 37 (23)
Wildtype - no. (%) / 14 (19) / 122 (75)
missing – no. (%) / 1 (1) / 4 (2)
NPM1mutated/FLT3-ITD negative / .98
low risk* – no. (%) / 26 (35) / 56 (34)
high risk* – no. (%) / 48 (65) / 104 (64)
missing – no. (%) / 0 (0) / 3 (2)
CEBPA / .14
mutated – no. (%) / 7 (10) / 27 (16)
Wildtype - no. (%) / 66 (89) / 133 (82)
missing – no. (%) / 1 (1) / 3 (2)
DNMT3A / .14
mutated – no. (%) / 25 (34) / 40 (25)
Wildtype - no. (%) / 48 (65) / 121 (74)
missing – no. (%) / 1 (1) / 2 (1)
WT1 / .6
mutated – no. (%) / 10 (14) / 18 (11)
Wildtype - no. (%) / 64 (86) / 144 (88)
missing – no. (%) / 0 (0) / 1 (1)
WT1 SNP rs16754 / .61
≥ 1 minor allele – no. (%) / 22 (30) / 43 (26)
missing – no. (%) / 0 (0) / 1 (1)
NRAS / .099
mutated – no. (%) / 6 (8) / 26 (16)
Wildtype - no. (%) / 66 (89) / 132 (81)
missing – no. (%) / 2 (3) / 5 (3)
IDH1 / .92
mutated – no. (%) / 7 (9) / 16 (10)
Wildtype - no. (%) / 67 (91) / 146 (89)
missing – no. (%) / 0 (0) / 1 (1)
IDH2 / .12
mutated – no. (%) / 5 (7) / 22 (14)
Wildtype - no. (%) / 69 (93) / 139 (85)
missing – no. (%) / 0 (0) / 2 (1)
BAALC expression / .89
median - (BAALC/ABL) / 4.67 / 4.38
range - (BAALC/ABL) / 0.168-262.75 / 0.026-806.36
missing data – no. (%) / 2 (3) / 3 (2)
ERG expression / <.001
median - (ERG x104/ABL) / 4.921 / 2.86
range - (ERG x104/ABL) / 0.364-27.53 / 0.007-160.57
missing data – no. (%) / 0 (0) / 2 (1)
MN1 expression / .33
median - (MN1x104/ABL) / 13 / 18.4
range - (MN1 x104/ABL) / 1.1-532 / 3.098-18570
missing data – no. (%) / 0 (0) / 3 (2)
WT1 expression / <.001
median - (WT1/ABL) / 2.44 / 0.7
range - (WT1/ABL) / 0.0032-181.79 / 0.0011-261.33
missing data – no. (%) / 0 (0) / 0 (0)
MLL5 expression / .13
median – (MLL5/ABL) / 34.2 / 39.47
range - (MLL5/ABL) / 10.49-231.02 / 6.377-498.02
missing data – no. (%) / 0 (0) / 0 (0)

Abbreviations: AML, acute myeloid leukemia; ECOG, performance status of the Eastern Cooperative Oncology Group; FAB, French-American-British classification of acute myeloid leukemia; FLT3-ITD, internal tandem duplication of the FLT3 gene; P, p-value from two-sided Chi-squared tests for categorical variables and from two-sided Mann-Whitney-U tests for continuous variables; SNP, single nucleotide polymorphism; WBC, white blood cell count.

* The high-risk molecular group is defined as either NPM1wildtype/FLT3-ITDnegative, or NPM1wildtype/FLT3-ITDpositive, or NPM1mutated/FLT3-ITDpositive. The low-risk molecular group is defined by the presence of an NPM1 mutation and the absence of FLT3-ITD.

Table S2. Comparison of pretreatment characteristics of 453 AML patients according to NUMB expression levels.

Characteristic / High NUMB expressers / Low NUMB expressers / P
(n = 114) / (n=339)
Age, years / .68
median / 46 / 46
range / 18-60 / 17-60
Sex / .11
male - no. (%) / 54 (47) / 190 (56)
female - no. (%) / 60 (53) / 149 (44)
ECOG performance status / .43
0 - no. (%) / 26 (23) / 83 (24)
1 - no. (%) / 67 (59) / 213 (63)
2 - no. (%) / 19 (16) / 41 (12)
Missing data - no. (%) / 2 (2) / 2 (1)
FAB-subtype / .018
M0 - no. (%) / 1 (1) / 10 (3)
M1 - no. (%) / 10 (9) / 70 (20)
M2 - no. (%) / 24 (21) / 84 (24)
M4 - no. (%) / 46 (40) / 113 (33)
M5 - no. (%) / 24 (21) / 39 (12)
M6 - no. (%) / 1 (1) / 9 (3)
M7 - no. (%) / 1 (1) / 2 (1)
missing data - no. (%) / 7 (6) / 12 (4)
Cytogenetic risk group / .53
Favorable – no. (%) / 16 (14) / 60 (18)
Intermediate – no. (%) / 83 (73) / 231 (68)
Adverse – no. (%) / 13 (11) / 46 (13)
missing data - no. (%) / 2 (2) / 2 (1)
FLT3-ITD – no. (%) / .56
mutated – no. (%) / 32 (28) / 88 (26)
WT – no. (%) / 79 (69) / 250 74)
missing – no. (%) / 3 (2) / 1 (0)
NPM1 / .59
mutated – no. (%) / 40 (35) / 111 (33)
WT – no. (%) / 72 (63) / 226 (66)
missing – no. (%) / 2 (2) / 2 (1)
NPM1mutated/FLT3-ITD negative / .059
low risk# – no. (%) / 29 (25) / 60 (18)
high risk* – no. (%) / 83 (73) / 279 (82)
missing – no. (%) / 2 (2) / 0 (0)
DNMT3A / .062
mutated – no. (%) / 26 (23) / 53 (16)
WT – no. (%) / 82 (72) / 276 (81)
missing – no. (%) / 6 (5) / 10 (3)
NRAS / .074
mutated – no. (%) / 19 (16) / 39 (11)
WT – no. (%) / 69 (61) / 246 (73)
missing – no. (%) / 26 (23) / 54 (16)
IDH1 / .019
mutated – no. (%) / 2 (2) / 27 (8)
WT – no. (%) / 85 (74) / 234 (69)
missing – no. (%) / 27 (24) / 78 (23)
IDH2 / .75
mutated – no. (%) / 7 (6) / 24 (7)
WT – no. (%) / 79 (69) / 235 (69)
missing – no. (%) / 28 (25) / 80 (24)
MLL5 Expression / .48
Mean copy number
(MLL5/ABL) / 6.4-336.9 / 8.5-498
Range
(MLL5/ABL) / 51.1 / 33.1
Missing data - no.(%) / 0 (0) / 2 (1)
Type of AML / .75
de novo - no. (%) / 13 (11) / 35 (10)
secondary - no. (%) / 101 (89) / 304 (90)
Peripheral blood blasts / .004
median (%) / 24 / 52
missing data – no. (%) / 5 (4) / 11 (3)
Bone marrow blasts / .79
median (%) / 80 / 80
missing data – no. (%) / 9 (8) / 21 (6)
WBC count / .24
median - (x109/l) / 29 / 17
range - (x109/l) / .7-271 / .5-284
missing data – no. (%) / 1 (1) / 1 (0)
Hemoglobin / .52
median – g/L / 8.9 / 9.1
range – g/L / 3.4-14.9 / 3-15.4
missing data – no. (%) / 3 (3) / 9 (3)
Platelet count / .94
median - (x109/l) / 49.5 / 45.5
range - (x109/l) / 7-316 / 2.6-624
missing data – no. (%) / 4 (4) / 9 (3)

Abbreviations: AML, acute myeloid leukemia; ECOG, performance status of the Eastern Cooperative Oncology Group; FAB, French-American-British classification of acute myeloid leukemia; FLT3-ITD, internal tandem duplication of the FLT3 gene; P, p-value from two-sided chi-squared tests for categorical variables and from two-sided Mann-Whitney-U tests for continuous variables.

Table S3. Comparison of pretreatment characteristics of CN-AML patients according to NUMB expression levels.

Characteristic / High NUMB expressers / Low NUMB expressers / P
(n=57) / (n=179)
Age, years / .97
Median / 48 / 47
range / 19-60 / 17-60
Sex / .3
male - no. (%) / 27 (47) / 99 (55)
female - no. (%) / 30 (53) / 80 (45)
FAB-Subtype / .001
M0 - no. (%) / 0 (0) / 7 (4)
M1 - no. (%) / 6 (11) / 40 (23)
M2 - no. (%) / 6 (11) / 45 (25)
M4 - no. (%) / 24 (42) / 59 (33)
M5 - no. (%) / 16 (28) / 18 (10)
M6 - no. (%) / 0 (0) / 6 (3)
M7 - no. (%) / 1 (1) / 2 (1)
missing data - no. (%) / 4 (7) / 2 (1)
Peripheral blood blasts / .3
median (%) / 21 / 59
missing data – no. (%) / 2 (4) / 9 (5)
Bone marrow blasts / .2
median (%) / 80 / 80
missing data – no. (%) / 5 (9) / 14 (8)
Type of AML / .62
de novo - no. (%) / 51 (89) / 164 (92)
secondary - no. (%) / 6 (11) / 15 (8)
WBC count / .43
median - (x109/l) / 30.5 / 25.11
range - (x109/l) / 1.2-185 / 0.5-239.4
missing data – no. (%) / 0 (0) / 0 (0)
Hemoglobin / .54
median – g/L / 9 / 9.1
range – g/L / 5.1-13.7 / 3-15.4
missing data – no. (%) / 2 (4) / 6 (3)
Platelet count / .43
median - (x109/l) / 85.5 / 45
range - (x109/l) / 9-316 / 4-483
missing data – no. (%) / 3 (5) / 6 (3)
ECOG performance status / .27
0 - no. (%) / 14 (25) / 37 (21)
1 - no. (%) / 39 (68) / 117 (65)
2 - no. (%) / 3 (5) / 23 (13)
missing data - no. (%) / 1 (2) / 2 (1)
FLT3-ITD – no. (%) / .9
mutated– no. (%) / 18 (32) / 61 (34)
Wildtype - no. (%) / 36 (63) / 117 (65)
missing – no. (%) / 3 (5) / 1 (1)
NPM1 / .12
mutated – no. (%) / 37 (65) / 98 (55)
Wildtype - no. (%) / 18 (32) / 79 (44)
missing – no. (%) / 2 (3) / 2 (1)
NPM1mutated/FLT3-ITD negative / .005
low risk* – no. (%) / 28 (49) / 54 (30)
high risk* – no. (%) / 27 (47) / 125 (70)
missing – no. (%) / 2 (4) / 0 (0)
CEBPA / .23
mutated – no. (%) / 5 (9) / 28 (15)
Wildtype - no. (%) / 49 (86) / 150 (84)
missing – no. (%) / 3 (5) / 1 (1)
DNMT3A / .003
mutated – no. (%) / 24 (42) / 41 (23)
Wildtype - no. (%) / 31 (54) / 137 (76)
missing – no. (%) / 2 (4) / 1 (1)
WT1 / .43
mutated – no. (%) / 5 (9) / 23 (13)
Wildtype - no. (%) / 51 (89) / 156 (87)
missing – no. (%) / 1 (2) / 0 (0)
WT1 SNP rs16754 / .26
≥ 1 minor allele – no. (%) / 12 (21) / 52 (29)
missing – no. (%) / 1 (2) / 0 (0)
NRAS / .054
mutated – no. (%) / 12 (21) / 20 (11)
Wildtype - no. (%) / 43 (75) / 154 (86)
missing – no. (%) / 2 (4) / 5 (3)
IDH1 / .088
mutated – no. (%) / 2 (3) / 20 (11)
Wildtype - no. (%) / 54 (95) / 159 (89)
missing – no. (%) / 1 (2) / 0 (0)
IDH2 / .83
mutated – no. (%) / 6 (10) / 21 (12)
Wildtype - no. (%) / 50 (88) / 157 (87)
missing – no. (%) / 1 (2) / 1 (1)
BAALC expression / .021
median - (BAALC/ABL) / 2.62 / 5.91
range - (BAALC/ABL) / 0.149-153.92 / 0.026-806.38
missing data – no. (%) / 1 (2) / 3 (2)
ERG expression / .022
median - (ERG x104/ABL) / 2.396 / 3.94
range - (ERG x104/ABL) / 0.124-20.73 / 0.007-160.56
missing data – no. (%) / 1 (2) / 1 (1)
MN1 expression / .006
median - (MN1 x104/ABL) / 9.50 / 19.79
range - (MN1 x104/ABL) / 1.13-443.37 / 0.31-18.57
missing data – no. (%) / 2 (4) / 0 (0)
WT1 expression / .001
median - (WT1/ABL) / 0.54 / 1.25
range - (WT1/ABL) / 0.0099-15.73 / 0.0011-261.33
missing data – no. (%) / 5 (9) / 6 (3)
MLL5 expression / <.001
median – (MLL5/ABL) / 52.62 / 33.13
range - (MLL5/ABL) / 6.38-205.3 / 9.96-498.02
missing data – no. (%) / 0 (0) / 0 (0)

Figure S1. Overall survival of all analyzed patients with AML according to MSI2 (A) and Numb (B) expression levels analyzed in MSI2 expression quartiles (log-rank test).

Figure S2. Overall survival in CN-AML patients with (A) FLT3-ITD and (B) wildtype FLT3 and NPM1 according to high vs. low MSI2 expression levels (log-rank test).

A B

Figure S3. (A) Overall and (B) relapse-free survival of all analyzed patients with AML according to high vs low NUMB expression levels (log-rank test).

A B

Figure S4. (A) Overall and (B) relapse-free survival of patients with CN-AML according to high vs low NUMB expression levels (log-rank test).

A B