Abstract

Children with Down syndrome (DS) up to the age of 4 years are at a 150-fold excess risk of developing myeloid leukemia (ML-DS). Approximately 4%-5% of newborns with DS develop transient myeloproliferative disorder (TMD). Blast cell structure and immunophenotype are similar in TMD and ML-DS. A mutation in the hematopoietic transcription factor GATA1 is present in almost all cases. Here, we show that simple techniques detect GATA1 mutations in the largest series of TMD (n = 134; 88%) and ML-DS (n = 103; 85%) cases tested. Furthermore, no significant difference in the mutational spectrum between the 2 disorders was seen. Thus, the type of GATA1 sequence mutation is not a reliable tool and is not prognostic of which patients with TMD are probable to develop ML-DS.

Introduction

Children with trisomy 21 (T21; Down syndrome [DS]) have ∼ 150-fold increased incidence of myeloid leukemia (ML-DS).1,2  Incidence of transient myeloproliferative disorder (TMD) is estimated at ∼ 4%-5% of neonates with DS.1,3  Approximately 20%-30% of these neonates develop ML-DS by 4 years of age.4,5  Both diseases are characterized by a clonal population of blasts, with similar immunophenotype and structure in blood and BM. However, TMD spontaneously regresses, whereas ML-DS is stably transformed.

In addition to T21, blast cells in TMD and ML-DS carry acquired mutations in the hematopoietic transcription factor GATA1.6-14  These mutations lead to expression of N-terminally truncated GATA1s protein. Mutations are detectable in disease but not in remission.3,6-14  Most reported mutations are found in GATA1 exon 2, including insertions, deletions, and point mutations. When TMD progresses to ML-DS, the same GATA1 mutation is usually present in blasts of both, showing their clonal relationship.5,14 

Debate exists about whether the type of GATA1 mutation determines progression to ML-DS.9,15-17  To examine this, we analyzed GATA1 mutations in 134 TMD and 103 ML-DS cases, the largest patient cohort reported to date. Of these 8 paired TMD and follow-up ML-DS samples were available. GATA1 mutations were detected in 226 patients (95%). The lower limit blast percentage for successful detection of GATA1 mutations was 0.5%. No difference was observed in types of mutation between patients with TMD and with ML-DS. Contrary to previous data,15  we did not detect specific GATA1 mutation types more commonly in ML-DS. Therefore, the type of GATA1 mutation in our series is not prognostic of which patients with TMD will progress to ML-DS.

Methods

Mutation detection DNA was prepared from peripheral blood or BM with the use of the DNeasy Blood and Tissue kit (QIAGEN). PCR was performed with primers and conditions outlined in supplemental Methods (available on the Blood Web site; see the Supplemental Materials link at the top of the online article). PCR amplicons were analyzed by denaturing high-performance liquid chromatography (WAVE; Transgenomic) and direct sequencing. In sample subsets with blast percentage < 1%, blasts were sorted before DNA extraction (supplemental Methods), or PCR product was cloned with the pGEM-T-Easy vector system 1 kit (Promega) and sequenced.

Statistical analysis was performed with the Fisher exact test.

Results and discussion

GATA1 mutation screening was performed in the central reference of Acute Myeloid Leukemia Berlin-Frankfurt-Münster Study group in Hannover, Germany, and the Weatherall Institute of Molecular Medicine, Oxford, United Kingdom (134 TMD and 103 ML-DS samples). The mean age of patients with TMD was 0.78 months (range, 0-11 months). Mean blast count was 42% (range, 0.5%-95%), white blood cell count was 60.79 × 109/L (range, 1-1193.3 × 109/L), hemoglobin level was 13.77 g/dL (range, 2-21.3 g/dL), and platelet count was 201.78 × 109/L (range, 12-1800 × 109/L; Table 1). The mean age of the patients with DS-ML was 20.19 months (range, 1-60 months). Mean blast count was 27.8% (range, 1%-96%), white blood cell count was 13.95 × 109/L (range, 1-200.3 × 109/L), hemoglobin level was 9.06 g/dL (range, 3.1-15.1 g/dL) and platelet count was 52.2 × 109/L (range, 1.5-257 × 109/L; Table 2).

Table 1

Summary of GATA1 mutations and patient characteristics for TMD samples

Patient no. Blast count, % of total nucleated count WAVE* Mutation type No. of nucleotides changed, bp Position of mutation† Predicted effect of mutation on RNA Age at diagnosis, mo WBC count, ×109/L Hb level, g/dL Plt count, ×109/L Kanezaki et al15  class Karyotype Outcome 
79 Ex2 Point 4549 Loss of start codon 107 15 140 1st Met 47, XY, +21c CCR 
2‡ 66 Ex2 Dup 20 4722 Frameshift and introduction of stop codon 70 13.3 352 PTC 1-3′ 47, XX, +21c ML-DS 
40 Ex2 Point 4767 Loss of splice acceptor site 1.3 14 12.7 461 Splice 47, XY, +21c CCR 
35 Ex2 Ins 4701 Frameshift and introduction of stop codon 16 21.3 41 PTC 1-5′ 47, XY, +21c CCR 
30 Ex2 Dup 101 4653 Frameshift and introduction of stop codon 0.57 8.2 122 PTC 1-5′ 47, XY, +21c CCR 
55 Ex2 Dup 19 4733 Frameshift and introduction of stop codon 0.57 75.9 6.3 113 PTC 1-3′ 47, XX, +21c CCR 
29 Ex2 Del 4638 Frameshift and introduction of stop codon 0.28 44.6 17.4 45 PTC 1-5′ 47, XX, +21c CCR 
92 Ex2 Dup 4768 Loss of splice acceptor site 0.28 120 18.8 505 Splice N/A CCR 
9‡ 40 Ex2 Point 4583 Nonsynonymous change in amino acid sequence introducing a stop codon 50 N/A N/A PTC 1-5′ N/A ML-DS 
10 80 Ex2 Dup 22 4735 Frameshift and introduction of stop codon 121 11.6 35 PTC 1-3′ 47, XY, +21 Thrombo-cytopaenia 
11 11 Ex2 Dup 17 4719 Frameshift and introduction of stop codon 38.2 18.7 132 PTC 1-3′ N/A CCR 
12 21 Ex2 Point 4768 Loss of splice acceptor site 19.4 10.8 52 Splice N/A Unknown 
13 Ex2 Del 4607 Frameshift and introduction of stop codon 0.71 9.5 12.7 54 PTC 1-5′ N/A Unknown 
14 60 Ex2 Ins 16 4721 Frameshift and introduction of stop codon 0.57 6.7 16.3 59 PTC 1-3′ N/A Died 
15 Ex2 Dup 22 4723 Frameshift and introduction of stop codon 0.71 12 14 88 PTC 1-3′ N/A Unknown 
16 35 Ex2 Del 26 4688 Frameshift and introduction of stop codon 39.3 13.8 335 PTC 1-5′ N/A CCR 
17 82 Ex2 Del 4458 Frameshift and introduction of stop codon 1.1 249 11.8 122 PTC 1-5′ 47, XY, +21c CCR 
18 75 Ex2 Ins 16 4706 Frameshift and introduction of stop codon N/A 31.6 16.2 44 PTC 1-5′ N/A CCR 
19 30 Ex2 Point 4550 Loss of start codon 11.5 13.3 72 1st Met 47, XY, +21c CCR 
20 N/A Ex2 Point 4596 Nonsynonymous change in amino acid sequence introducing a stop codon N/A N/A N/A N/A PTC 1-5′ N/A Died 
21 N/A Ex2 Point 4766 Loss of splice acceptor site N/A N/A N/A Splice N/A Unknown 
22 25 Ex2 Ins 4709 Frameshift and introduction of stop codon 33.7 14.8 35 PTC 1-5′ N/A Unknown 
23 45 Ex2 Substitution 4744 Nonsynonymous change in amino acid sequence introducing a stop codon 2.7 20 51 PTC 1-3′ 47, XY, der(11)t(q23p15) t(1:11)(q23;q85), +21c CCR 
24 50 Ex2 Del 132 4671 Loss of splice acceptor site 58 N/A N/A Splice 47, XY, +21c CCR 
25 76 Ex2 Ins 4661 Frameshift and introduction of stop codon 124.9 28 PTC 1-5′ 47, XY, +21c CCR 
26 25 Ex2 Del 4662 Frameshift and introduction of stop codon 39.6 19.8 22 PTC 1-5′ 47, XY, +21c CCR 
27‡ 18 Ex2 Dup 4717 Frameshift and introduction of stop codon 101 13.3 21 PTC 1-5′ 47, XY, t(5;11)(p15;q13), +21c ML-DS 
28 70 Ex2 Del 4684 Frameshift and introduction of stop codon 200 7.6 87 PTC 1-5′ N/A CCR 
29 62 Ex2 Ins 4643 Frameshift and introduction of stop codon N/A N/A N/A PTC 1-5′ N/A Unknown 
30 11.4 Ex2 Dup 4732 Frameshift and introduction of stop codon 33.5 9.1 695 PTC 1-3′ 47, XY, +21c Unknown 
31 29 Ex2 Point 4670 Nonsynonymous change in amino acid sequence introducing a stop codon 21.7 13 24 PTC 1-5′ N/A Unknown 
32 60 Ex2 Del 4697 Frameshift and introduction of stop codon 28 weeks gestation 40 5.1 54 PTC 1-5′ N/A Unknown 
33 79 N/D Del 4638 Frameshift and introduction of stop codon 198.2 10.6 312 PTC 1-5′ N/A Unknown 
34 66 N/D Dup 40 4740 Frameshift and introduction of stop codon 0.25 9.8 12.7 60 PTC 1-3′ N/A Unknown 
35 60 N/D Dup 10 4710 Frameshift and introduction of stop codon 0.25 46.6 20.4 89 PTC 1-5′ N/A Unknown 
36‡ 35.5 N/D Ins 14 4707 Frameshift and introduction of stop codon 55.6 17.3 570 PTC 1-5′ N/A ML-DS 
37 54 N/D Point 4768 Loss of splice acceptor site 55.6 17.3 570 Splice N/A Unknown 
38 23 N/D Point 4767 Synonymous mutation 1.25 19.2 6.7 125 Unknown N/A Unknown 
39‡ 84 N/D Del 4604 Frameshift and introduction of stop codon 1193.3 14.7 255 PTC 1-5′ N/A ML-DS 
40 10 N/D Del + Ins 2 + 1 4770 + 4774 Loss of splice donor site 9.1 9.6 77 Splice N/A Unknown 
41‡ 88 N/D Del 4680 Frameshift and introduction of stop codon 232 15.6 254 PTC 1-5′ N/A ML-DS 
42 75 N/D Del + Ins 1 + 2 4679 Frameshift and introduction of stop codon 172.8 16.2 101 PTC 1-5′ N/A Unknown 
43 14 N/D Point 4734 Unknown 0.25 16.9 14.9 353 Unknown N/A Unknown 
44 13.5 N/D Dup 36 4737 Frameshift and introduction of stop codon 0.5 3.2 11.1 16 PTC 1-3′ N/A Unknown 
45 31 N/D Dup 19 4722 Frameshift and introduction of stop codon 28.6 16.2 57 PTC 1-3′ N/A Unknown 
46 43 N/D Ins + Dup 2 + 13 4723 Frameshift and introduction of stop codon 61 17.6 149 PTC 1-3′ N/A Unknown 
47 69 N/D Point 4550 Loss of start codon 34 15 26 1st Met N/A Unknown 
48 10 N/D Dup 21 4723 Frameshift and introduction of stop codon 0.25 7.7 13.2 78 PTC 1-3′ N/A Unknown 
49 78 N/D Del + Ins 19 + 17 4698 Frameshift and introduction of stop codon 68.6 10.9 212 PTC 1-5′ N/A Unknown 
50 34 N/D Ins 4713 Frameshift and introduction of stop codon 0.25 6.2 16.9 25 PTC 1-5′ N/A Unknown 
51 60 N/D Del 4638 Frameshift and introduction of stop codon 51.2 14.8 159 PTC 1-5′ N/A Unknown 
52 75 N/D Point 4747 Nonsynonymous change in amino acid sequence introducing a stop codon N/A N/A N/A PTC 1-3′ N/A Unknown 
53 18.5 N/D Del 162 4540 Loss of start codon 17.3 14 173 1st Met N/A Unknown 
54 N/D Del + Ins 2 + 3 4698 Frameshift and introduction of sop codon 0.25 57.5 10.6 36 PTC 1-5′ N/A Unknown 
55 N/D Ins + Dup 3 + 36 4724 Frameshift and introduction of stop codon 0.25 16 18.5 48 PTC 1-3′ N/A Unknown 
56 66.5 N/D Del 4638 Frameshift and introduction of stop codon 0.75 62.9 12.3 93 PTC 1-5′ N/A Unknown 
57‡ 75 N/D Dup 13 4744 Frameshift and introduction of stop codon 149.1 16.9 131 PTC 1-3′ 47, XY,+21.ish 21qter(D21S1446 × 3) ML-DS 
58 85 N/D Point 4762 Nonsynonymous change in amino acid sequence introducing a stop codon 46.4 20.6 197 PTC 1-3′ N/A Unknown 
59 45 N/D Del + Ins 7 + 10 4760 Frameshift and introduction of stop codon 0.5 78.3 15 1800 PTC 1-3′ N/A Unknown 
60 38 N/D Ins + Dup 3 + 9 4740 Frameshift and introduction of stop codon 0.25 32.9 11.9 220 PTC 1-3′ N/A Unknown 
61‡ 64.5 N/D Dup 18 4733 Frameshift and introduction of stop codon 69.4 15.4 1178 PTC 1-3′ N/A ML-DS 
62 58 N/D Del 4698 Frameshift and introduction of stop codon 0.25 24.9 15.6 154 PTC 1-5′ N/A Unknown 
63 34 N/D Del 4690 Frameshift and introduction of stop codon 27.2 17 134 PTC 1-5′ N/A Unknown 
64 85 N/D Del 4604 Frameshift and introduction of stop codon 22 16.3 108 PTC 1-5′ N/A Unknown 
65 11 N/D Point 4549 Loss of s 7.4 12.3 241 1st Met N/A Unknown 
66 32 N/D Ins 4654 Frameshift and introduction of stop codon 31.6 16.4 158 PTC 1-5′ N/A Unknown 
67 3.5 N/D Point 4597 Nonsynonymous change in amino acid sequence 8.4 9.3 105 Unknown N/A Unknown 
68 22 N/D Del 34 4693 Frameshift and introduction of stop codon 0.25 17.3 13.1 270 PTC 1-5′ N/A Unknown 
69‡ 87 N/D Point 4552 Nonsynonymous change in amino acid sequence 0.25 133.9 13.8 32.9 Unknown N/A ML-DS 
70 5.5 N/D Point 4768 Loss of splice acceptor site 23.83 9.9 154 Splice N/A Unknown 
71 32 N/D Ins + Dup 1 + 12 4742 Frameshift and introduction of stop codon 27.6 14.6 46 PTC 1-3′ N/A Unknown 
72 91 N/D Ins 4647 Frameshift and introduction of stop codon 10.6 10.7 294 PTC 1-5′ N/A Unknown 
73 16.5 N/D Del 4653 Frameshift and introduction of stop codon 0.75 8.9 12.5 120 PTC 1-5′ N/A Unknown 
74 14 N/D Dup 19 4722 Frameshift and introduction of stop codon 11 3.39 10.5 91 PTC 1-3′ N/A Unknown 
75 78 N/D Del 4766 Loss of splice acceptor site 91.2 13.6 33 Splice 47, XY, t(21;21) (Q10;Q10) +21c Unknown 
76 60.5 N/D Point 4734 Nonsynonymous change in amino acid sequence 25.4 13.9 98 Unknown N/A Unknown 
77 16 N/D Dup 4734 Frameshift and introduction of stop codon 0.5 8.5 10.8 52 PTC 1-3′ N/A Unknown 
78 12 N/D Del 51 4706 Frameshift and introduction of stop codon 23.2 14.2 379 PTC 1-5′ N/A Unknown 
79 95 N/D Ins + Dup 6 + 7 4708 Frameshift and introduction of stop codon 410 11.1 261 PTC 1-5′ N/A Unknown 
80 86 N/D Point 4633 Nonsynonymous change in amino acid sequence 107.5 13.1 70 Unknown 47,XX,der(6)del(6) (p21)del(6)(q24, +der(6)del(6)(p12p22)t (6;10)(q16;q22),der(10) t(10;12)(q21;p12), der(11)t(11;12) (p15;q11),−12,+21c[17], 47,XX,+21c[2] Unknown 
81‡ 68 N/D Point 4769 Loss of splice acceptor site 100 11.5 326 Splice N/A ML-DS 
82 72 N/D Del 12 4733 Frameshift and introduction of stop codon 0.5 49.8 14.2 43 PTC 1-3′ N/A Unknown 
83 30 N/D Del 4741 Frameshift and introduction of top codon 52 13.9 71 PTC 1-3′ N/A Unknown 
84 35 N/D Ins 4654 Frameshift and introduction of stop codon 28 12.8 274 PTC 1-5′ N/A Unknown 
85 78 N/D Substitution 40 4545 Unknown 72.7 19.4 606 Unknown N/A Unknown 
86 79 N/D Ins 4736 Frameshift and introduction of stop codon 100 10.9 70 PTC 1-3′ N/A Unknown 
87 46.5 N/D Del 4637 Frameshift and introduction of stop codon 0.25 23.3 17.5 80 PTC 1-5′ N/A Unknown 
88‡ 43 N/D Del 4638 Frameshift and introduction of stop codon 0.25 N/A N/A N/A PTC 1-5′ 47,XY,+21c[15] ML-DS 
89 15 N/D Ins 4725 Frameshift and introduction of stop codon 0.5 11 24.2 158 PTC 1-3′ N/A Unknown 
90 16.5 N/D Del 4604 Frameshift and introduction of stop codon 0.25 24.2 13.2 78 PTC 1-5′ N/A Unknown 
91‡ 66 N/D Dup 4723 Frameshift and introduction of stop codon 26.1 16.5 48 PTC 1-3′ 47,XX,+21c ML-DS 
92 38.5 N/D Dup 43 4742 Frameshift and introduction of stop codon 0.5 20.2 12.3 12 PTC 1-3′ N/A Unknown 
93 58 N/D Point 4768 Loss of splice acceptor site 20.7 14.1 901 Splice N/A Unknown 
94 N/D Point + Point 1 + 1 92 + 126 Unknown 0.25 5.6 14.3 67 Unknown N/A Unknown 
95 83 N/D Del + Del 15 + 1 4888 + 4907 Frameshift and introduction of stop codon 0.25 114.7 16.6 141 PTC 2 N/A Unknown 
96 20 N/D Dup 4714 Frameshift and introduction of stop codon 14.12 18.6 15 PTC 1-5′ N/A Unknown 
97 43 N/D Point 4769 Loss of splice acceptor site 49.8 17 209 Splice N/A Unknown 
98 49.9 N/D Del 4638 Frameshift and introduction of stop codon 0.5 7.06 12 64 PTC 1-5′ N/A Unknown 
99 61 N/D Dup 12 4720 Frameshift and introduction of stop codon 0.75 91.6 16.6 150 PTC 1-3′ N/A Unknown 
100 53.5 N/D Ins 4598 Frameshift and introduction of stop codon 51 18.3 56 PTC 1-5′ N/A Unknown 
101 34 N/D Del 13 4706 Frameshift and introduction of stop codon 15.4 17.1 62 PTC 1-5′ N/A Unknown 
102 25 N/D Ins 4753 Frameshift and introduction of stop codon 19.9 18.3 115 PTC 1-3′ N/A Unknown 
103 60 N/D Dup 4707 Frameshift and introduction of stop codon 23.8 12.9 265 PTC 1-5′ N/A Unknown 
104 51 N/D Point 4748 Synonymous mutation 16.6 12.5 567 Unknown 47,XX,+21[5]/47,XX,der(7), +21[2]  
105 15.5 N/D Del + Ins 3 + 2 4742 Frameshift and introduction of stop codon 0.25 64.4 13.8 547 PTC 1-3′ N/A Unknown 
106 50 N/D Dup 19 4722 Frameshift and introduction of stop codon 43.5 15.4 128 PTC 1-3′ N/A Known 
107 21 N/D Dup 11 4735 Frameshift and introduction of stop codon 10.7 45 PTC 1-3′ N/A Unknown 
108 N/A N/D Ins 4731 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC -3′ N/A Unknown 
109 22 N/D Point 4769 Loss of splice acceptor site 1.5 74.7 554 Splice N/A Unknown 
110 57.5 N/D Dup 105 4698 Nonsynonymous change in amino acid sequence introducing a stop codon 64.3 11.8 427 PTC 1-5′ N/A Unknown 
111 62 N/D Del 12 4764 Loss of splice acceptor site 1.5 21.9 6.7 102 Splice 47,XY,+21c Unknown 
112 45 N/D Del 4672 Frameshift and introduction of stop codon N/A N/A N/A PTC 1-5′ N/A Unknown 
113 73 N/D Dup 4706 Frameshift and introduction of stop codon 195 12 150 PTC 1-5′ N/A Unknown 
114 N/A N/D Del 4586 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-5′ N/A Unknown 
115 77 N/D Dup 10 4719 Frameshift and introduction of stop codon 170 10.6 655 PTC 1-3′ N/A Unknown 
116‡ 0.5 N/D Ins 4733 Frameshift and introduction of stop codon 1.5 89 PTC 1-3′ 48, XY, +Y(q12),+11, +21c[8]/47, XY,+21c[7] ML-DS 
117 30 Ex2 Not found Unknown Unknown N/A 4.2 12.9 25 Unknown 47, XX, del(16), +21c Unknown 
118 10 Ex2 Not found Unknown Unknown N/A 20.4 13 119 Unknown N/A Unknown 
119 Ex2 Not found Unknown Unknown N/A 0.38 21.4 15 218 Unknown 47, XY, +21c Unknown 
120 Ex2 Not found Unknown Unknown N/A 20.7 16.1 63 Unknown 47, XY, +21c Unknown 
121 Ex2 Not found Unknown Unknown N/A 6.86 6.4 83 Unknown 47, XY, +21c Unknown 
122 30 Ex2 Not found Unknown Unknown N/A 2.5 13 1000 Unknown N/A Unknown 
123 80 Ex2 Not found Unknown Unknown N/A 265.3 18 353 Unknown N/A Unknown 
124 20 Ex2 Not found Unknown Unknown N/A 0.5 13.4 11.9 371 Unknown 47, XY, +21c Unknown 
125 39 Ex2 Not found Unknown Unknown N/A 12.5 16.1 193 Unknown N/A Unknown 
126 35 Ex 3.1 Del 16 4871 Frameshift and introduction of stop codon 33.5 9.1 695 PTC 2 N/A Unknown 
127 30 Ex 3.1 Dup 4778 Frameshift and introduction of stop codon 21.7 13 24 PTC 2 N/A Unknown 
128 13.5 N/D Not found Unknown Unknown N/A 0.25 8.91 18.4 46 Unknown 47,XX,+21c[17] CCR 
129 N/D Not found Unknown Unknown N/A 10.6 18.9 65 Unknown N/A CCR 
130 N/D Not found Unknown Unknown N/A 0.75 8.3 8.1 100 Unknown N/A Unknown 
131 N/D Not found Unknown Unknown N/A 22.3 21 60 Unknown N/A Unknown 
132 N/D Not found Unknown Unknown N/A 0.5 15.2 20.8 276 Unknown N/A Unknown 
133 12 N/D Not found Unknown Unknown N/A N/A N/A N/A Unknown N/A Unknown 
134 N/D Not found Unknown Unknown N/A 4.15 13 100 Unknown N/A Unknown 
Patient no. Blast count, % of total nucleated count WAVE* Mutation type No. of nucleotides changed, bp Position of mutation† Predicted effect of mutation on RNA Age at diagnosis, mo WBC count, ×109/L Hb level, g/dL Plt count, ×109/L Kanezaki et al15  class Karyotype Outcome 
79 Ex2 Point 4549 Loss of start codon 107 15 140 1st Met 47, XY, +21c CCR 
2‡ 66 Ex2 Dup 20 4722 Frameshift and introduction of stop codon 70 13.3 352 PTC 1-3′ 47, XX, +21c ML-DS 
40 Ex2 Point 4767 Loss of splice acceptor site 1.3 14 12.7 461 Splice 47, XY, +21c CCR 
35 Ex2 Ins 4701 Frameshift and introduction of stop codon 16 21.3 41 PTC 1-5′ 47, XY, +21c CCR 
30 Ex2 Dup 101 4653 Frameshift and introduction of stop codon 0.57 8.2 122 PTC 1-5′ 47, XY, +21c CCR 
55 Ex2 Dup 19 4733 Frameshift and introduction of stop codon 0.57 75.9 6.3 113 PTC 1-3′ 47, XX, +21c CCR 
29 Ex2 Del 4638 Frameshift and introduction of stop codon 0.28 44.6 17.4 45 PTC 1-5′ 47, XX, +21c CCR 
92 Ex2 Dup 4768 Loss of splice acceptor site 0.28 120 18.8 505 Splice N/A CCR 
9‡ 40 Ex2 Point 4583 Nonsynonymous change in amino acid sequence introducing a stop codon 50 N/A N/A PTC 1-5′ N/A ML-DS 
10 80 Ex2 Dup 22 4735 Frameshift and introduction of stop codon 121 11.6 35 PTC 1-3′ 47, XY, +21 Thrombo-cytopaenia 
11 11 Ex2 Dup 17 4719 Frameshift and introduction of stop codon 38.2 18.7 132 PTC 1-3′ N/A CCR 
12 21 Ex2 Point 4768 Loss of splice acceptor site 19.4 10.8 52 Splice N/A Unknown 
13 Ex2 Del 4607 Frameshift and introduction of stop codon 0.71 9.5 12.7 54 PTC 1-5′ N/A Unknown 
14 60 Ex2 Ins 16 4721 Frameshift and introduction of stop codon 0.57 6.7 16.3 59 PTC 1-3′ N/A Died 
15 Ex2 Dup 22 4723 Frameshift and introduction of stop codon 0.71 12 14 88 PTC 1-3′ N/A Unknown 
16 35 Ex2 Del 26 4688 Frameshift and introduction of stop codon 39.3 13.8 335 PTC 1-5′ N/A CCR 
17 82 Ex2 Del 4458 Frameshift and introduction of stop codon 1.1 249 11.8 122 PTC 1-5′ 47, XY, +21c CCR 
18 75 Ex2 Ins 16 4706 Frameshift and introduction of stop codon N/A 31.6 16.2 44 PTC 1-5′ N/A CCR 
19 30 Ex2 Point 4550 Loss of start codon 11.5 13.3 72 1st Met 47, XY, +21c CCR 
20 N/A Ex2 Point 4596 Nonsynonymous change in amino acid sequence introducing a stop codon N/A N/A N/A N/A PTC 1-5′ N/A Died 
21 N/A Ex2 Point 4766 Loss of splice acceptor site N/A N/A N/A Splice N/A Unknown 
22 25 Ex2 Ins 4709 Frameshift and introduction of stop codon 33.7 14.8 35 PTC 1-5′ N/A Unknown 
23 45 Ex2 Substitution 4744 Nonsynonymous change in amino acid sequence introducing a stop codon 2.7 20 51 PTC 1-3′ 47, XY, der(11)t(q23p15) t(1:11)(q23;q85), +21c CCR 
24 50 Ex2 Del 132 4671 Loss of splice acceptor site 58 N/A N/A Splice 47, XY, +21c CCR 
25 76 Ex2 Ins 4661 Frameshift and introduction of stop codon 124.9 28 PTC 1-5′ 47, XY, +21c CCR 
26 25 Ex2 Del 4662 Frameshift and introduction of stop codon 39.6 19.8 22 PTC 1-5′ 47, XY, +21c CCR 
27‡ 18 Ex2 Dup 4717 Frameshift and introduction of stop codon 101 13.3 21 PTC 1-5′ 47, XY, t(5;11)(p15;q13), +21c ML-DS 
28 70 Ex2 Del 4684 Frameshift and introduction of stop codon 200 7.6 87 PTC 1-5′ N/A CCR 
29 62 Ex2 Ins 4643 Frameshift and introduction of stop codon N/A N/A N/A PTC 1-5′ N/A Unknown 
30 11.4 Ex2 Dup 4732 Frameshift and introduction of stop codon 33.5 9.1 695 PTC 1-3′ 47, XY, +21c Unknown 
31 29 Ex2 Point 4670 Nonsynonymous change in amino acid sequence introducing a stop codon 21.7 13 24 PTC 1-5′ N/A Unknown 
32 60 Ex2 Del 4697 Frameshift and introduction of stop codon 28 weeks gestation 40 5.1 54 PTC 1-5′ N/A Unknown 
33 79 N/D Del 4638 Frameshift and introduction of stop codon 198.2 10.6 312 PTC 1-5′ N/A Unknown 
34 66 N/D Dup 40 4740 Frameshift and introduction of stop codon 0.25 9.8 12.7 60 PTC 1-3′ N/A Unknown 
35 60 N/D Dup 10 4710 Frameshift and introduction of stop codon 0.25 46.6 20.4 89 PTC 1-5′ N/A Unknown 
36‡ 35.5 N/D Ins 14 4707 Frameshift and introduction of stop codon 55.6 17.3 570 PTC 1-5′ N/A ML-DS 
37 54 N/D Point 4768 Loss of splice acceptor site 55.6 17.3 570 Splice N/A Unknown 
38 23 N/D Point 4767 Synonymous mutation 1.25 19.2 6.7 125 Unknown N/A Unknown 
39‡ 84 N/D Del 4604 Frameshift and introduction of stop codon 1193.3 14.7 255 PTC 1-5′ N/A ML-DS 
40 10 N/D Del + Ins 2 + 1 4770 + 4774 Loss of splice donor site 9.1 9.6 77 Splice N/A Unknown 
41‡ 88 N/D Del 4680 Frameshift and introduction of stop codon 232 15.6 254 PTC 1-5′ N/A ML-DS 
42 75 N/D Del + Ins 1 + 2 4679 Frameshift and introduction of stop codon 172.8 16.2 101 PTC 1-5′ N/A Unknown 
43 14 N/D Point 4734 Unknown 0.25 16.9 14.9 353 Unknown N/A Unknown 
44 13.5 N/D Dup 36 4737 Frameshift and introduction of stop codon 0.5 3.2 11.1 16 PTC 1-3′ N/A Unknown 
45 31 N/D Dup 19 4722 Frameshift and introduction of stop codon 28.6 16.2 57 PTC 1-3′ N/A Unknown 
46 43 N/D Ins + Dup 2 + 13 4723 Frameshift and introduction of stop codon 61 17.6 149 PTC 1-3′ N/A Unknown 
47 69 N/D Point 4550 Loss of start codon 34 15 26 1st Met N/A Unknown 
48 10 N/D Dup 21 4723 Frameshift and introduction of stop codon 0.25 7.7 13.2 78 PTC 1-3′ N/A Unknown 
49 78 N/D Del + Ins 19 + 17 4698 Frameshift and introduction of stop codon 68.6 10.9 212 PTC 1-5′ N/A Unknown 
50 34 N/D Ins 4713 Frameshift and introduction of stop codon 0.25 6.2 16.9 25 PTC 1-5′ N/A Unknown 
51 60 N/D Del 4638 Frameshift and introduction of stop codon 51.2 14.8 159 PTC 1-5′ N/A Unknown 
52 75 N/D Point 4747 Nonsynonymous change in amino acid sequence introducing a stop codon N/A N/A N/A PTC 1-3′ N/A Unknown 
53 18.5 N/D Del 162 4540 Loss of start codon 17.3 14 173 1st Met N/A Unknown 
54 N/D Del + Ins 2 + 3 4698 Frameshift and introduction of sop codon 0.25 57.5 10.6 36 PTC 1-5′ N/A Unknown 
55 N/D Ins + Dup 3 + 36 4724 Frameshift and introduction of stop codon 0.25 16 18.5 48 PTC 1-3′ N/A Unknown 
56 66.5 N/D Del 4638 Frameshift and introduction of stop codon 0.75 62.9 12.3 93 PTC 1-5′ N/A Unknown 
57‡ 75 N/D Dup 13 4744 Frameshift and introduction of stop codon 149.1 16.9 131 PTC 1-3′ 47, XY,+21.ish 21qter(D21S1446 × 3) ML-DS 
58 85 N/D Point 4762 Nonsynonymous change in amino acid sequence introducing a stop codon 46.4 20.6 197 PTC 1-3′ N/A Unknown 
59 45 N/D Del + Ins 7 + 10 4760 Frameshift and introduction of stop codon 0.5 78.3 15 1800 PTC 1-3′ N/A Unknown 
60 38 N/D Ins + Dup 3 + 9 4740 Frameshift and introduction of stop codon 0.25 32.9 11.9 220 PTC 1-3′ N/A Unknown 
61‡ 64.5 N/D Dup 18 4733 Frameshift and introduction of stop codon 69.4 15.4 1178 PTC 1-3′ N/A ML-DS 
62 58 N/D Del 4698 Frameshift and introduction of stop codon 0.25 24.9 15.6 154 PTC 1-5′ N/A Unknown 
63 34 N/D Del 4690 Frameshift and introduction of stop codon 27.2 17 134 PTC 1-5′ N/A Unknown 
64 85 N/D Del 4604 Frameshift and introduction of stop codon 22 16.3 108 PTC 1-5′ N/A Unknown 
65 11 N/D Point 4549 Loss of s 7.4 12.3 241 1st Met N/A Unknown 
66 32 N/D Ins 4654 Frameshift and introduction of stop codon 31.6 16.4 158 PTC 1-5′ N/A Unknown 
67 3.5 N/D Point 4597 Nonsynonymous change in amino acid sequence 8.4 9.3 105 Unknown N/A Unknown 
68 22 N/D Del 34 4693 Frameshift and introduction of stop codon 0.25 17.3 13.1 270 PTC 1-5′ N/A Unknown 
69‡ 87 N/D Point 4552 Nonsynonymous change in amino acid sequence 0.25 133.9 13.8 32.9 Unknown N/A ML-DS 
70 5.5 N/D Point 4768 Loss of splice acceptor site 23.83 9.9 154 Splice N/A Unknown 
71 32 N/D Ins + Dup 1 + 12 4742 Frameshift and introduction of stop codon 27.6 14.6 46 PTC 1-3′ N/A Unknown 
72 91 N/D Ins 4647 Frameshift and introduction of stop codon 10.6 10.7 294 PTC 1-5′ N/A Unknown 
73 16.5 N/D Del 4653 Frameshift and introduction of stop codon 0.75 8.9 12.5 120 PTC 1-5′ N/A Unknown 
74 14 N/D Dup 19 4722 Frameshift and introduction of stop codon 11 3.39 10.5 91 PTC 1-3′ N/A Unknown 
75 78 N/D Del 4766 Loss of splice acceptor site 91.2 13.6 33 Splice 47, XY, t(21;21) (Q10;Q10) +21c Unknown 
76 60.5 N/D Point 4734 Nonsynonymous change in amino acid sequence 25.4 13.9 98 Unknown N/A Unknown 
77 16 N/D Dup 4734 Frameshift and introduction of stop codon 0.5 8.5 10.8 52 PTC 1-3′ N/A Unknown 
78 12 N/D Del 51 4706 Frameshift and introduction of stop codon 23.2 14.2 379 PTC 1-5′ N/A Unknown 
79 95 N/D Ins + Dup 6 + 7 4708 Frameshift and introduction of stop codon 410 11.1 261 PTC 1-5′ N/A Unknown 
80 86 N/D Point 4633 Nonsynonymous change in amino acid sequence 107.5 13.1 70 Unknown 47,XX,der(6)del(6) (p21)del(6)(q24, +der(6)del(6)(p12p22)t (6;10)(q16;q22),der(10) t(10;12)(q21;p12), der(11)t(11;12) (p15;q11),−12,+21c[17], 47,XX,+21c[2] Unknown 
81‡ 68 N/D Point 4769 Loss of splice acceptor site 100 11.5 326 Splice N/A ML-DS 
82 72 N/D Del 12 4733 Frameshift and introduction of stop codon 0.5 49.8 14.2 43 PTC 1-3′ N/A Unknown 
83 30 N/D Del 4741 Frameshift and introduction of top codon 52 13.9 71 PTC 1-3′ N/A Unknown 
84 35 N/D Ins 4654 Frameshift and introduction of stop codon 28 12.8 274 PTC 1-5′ N/A Unknown 
85 78 N/D Substitution 40 4545 Unknown 72.7 19.4 606 Unknown N/A Unknown 
86 79 N/D Ins 4736 Frameshift and introduction of stop codon 100 10.9 70 PTC 1-3′ N/A Unknown 
87 46.5 N/D Del 4637 Frameshift and introduction of stop codon 0.25 23.3 17.5 80 PTC 1-5′ N/A Unknown 
88‡ 43 N/D Del 4638 Frameshift and introduction of stop codon 0.25 N/A N/A N/A PTC 1-5′ 47,XY,+21c[15] ML-DS 
89 15 N/D Ins 4725 Frameshift and introduction of stop codon 0.5 11 24.2 158 PTC 1-3′ N/A Unknown 
90 16.5 N/D Del 4604 Frameshift and introduction of stop codon 0.25 24.2 13.2 78 PTC 1-5′ N/A Unknown 
91‡ 66 N/D Dup 4723 Frameshift and introduction of stop codon 26.1 16.5 48 PTC 1-3′ 47,XX,+21c ML-DS 
92 38.5 N/D Dup 43 4742 Frameshift and introduction of stop codon 0.5 20.2 12.3 12 PTC 1-3′ N/A Unknown 
93 58 N/D Point 4768 Loss of splice acceptor site 20.7 14.1 901 Splice N/A Unknown 
94 N/D Point + Point 1 + 1 92 + 126 Unknown 0.25 5.6 14.3 67 Unknown N/A Unknown 
95 83 N/D Del + Del 15 + 1 4888 + 4907 Frameshift and introduction of stop codon 0.25 114.7 16.6 141 PTC 2 N/A Unknown 
96 20 N/D Dup 4714 Frameshift and introduction of stop codon 14.12 18.6 15 PTC 1-5′ N/A Unknown 
97 43 N/D Point 4769 Loss of splice acceptor site 49.8 17 209 Splice N/A Unknown 
98 49.9 N/D Del 4638 Frameshift and introduction of stop codon 0.5 7.06 12 64 PTC 1-5′ N/A Unknown 
99 61 N/D Dup 12 4720 Frameshift and introduction of stop codon 0.75 91.6 16.6 150 PTC 1-3′ N/A Unknown 
100 53.5 N/D Ins 4598 Frameshift and introduction of stop codon 51 18.3 56 PTC 1-5′ N/A Unknown 
101 34 N/D Del 13 4706 Frameshift and introduction of stop codon 15.4 17.1 62 PTC 1-5′ N/A Unknown 
102 25 N/D Ins 4753 Frameshift and introduction of stop codon 19.9 18.3 115 PTC 1-3′ N/A Unknown 
103 60 N/D Dup 4707 Frameshift and introduction of stop codon 23.8 12.9 265 PTC 1-5′ N/A Unknown 
104 51 N/D Point 4748 Synonymous mutation 16.6 12.5 567 Unknown 47,XX,+21[5]/47,XX,der(7), +21[2]  
105 15.5 N/D Del + Ins 3 + 2 4742 Frameshift and introduction of stop codon 0.25 64.4 13.8 547 PTC 1-3′ N/A Unknown 
106 50 N/D Dup 19 4722 Frameshift and introduction of stop codon 43.5 15.4 128 PTC 1-3′ N/A Known 
107 21 N/D Dup 11 4735 Frameshift and introduction of stop codon 10.7 45 PTC 1-3′ N/A Unknown 
108 N/A N/D Ins 4731 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC -3′ N/A Unknown 
109 22 N/D Point 4769 Loss of splice acceptor site 1.5 74.7 554 Splice N/A Unknown 
110 57.5 N/D Dup 105 4698 Nonsynonymous change in amino acid sequence introducing a stop codon 64.3 11.8 427 PTC 1-5′ N/A Unknown 
111 62 N/D Del 12 4764 Loss of splice acceptor site 1.5 21.9 6.7 102 Splice 47,XY,+21c Unknown 
112 45 N/D Del 4672 Frameshift and introduction of stop codon N/A N/A N/A PTC 1-5′ N/A Unknown 
113 73 N/D Dup 4706 Frameshift and introduction of stop codon 195 12 150 PTC 1-5′ N/A Unknown 
114 N/A N/D Del 4586 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-5′ N/A Unknown 
115 77 N/D Dup 10 4719 Frameshift and introduction of stop codon 170 10.6 655 PTC 1-3′ N/A Unknown 
116‡ 0.5 N/D Ins 4733 Frameshift and introduction of stop codon 1.5 89 PTC 1-3′ 48, XY, +Y(q12),+11, +21c[8]/47, XY,+21c[7] ML-DS 
117 30 Ex2 Not found Unknown Unknown N/A 4.2 12.9 25 Unknown 47, XX, del(16), +21c Unknown 
118 10 Ex2 Not found Unknown Unknown N/A 20.4 13 119 Unknown N/A Unknown 
119 Ex2 Not found Unknown Unknown N/A 0.38 21.4 15 218 Unknown 47, XY, +21c Unknown 
120 Ex2 Not found Unknown Unknown N/A 20.7 16.1 63 Unknown 47, XY, +21c Unknown 
121 Ex2 Not found Unknown Unknown N/A 6.86 6.4 83 Unknown 47, XY, +21c Unknown 
122 30 Ex2 Not found Unknown Unknown N/A 2.5 13 1000 Unknown N/A Unknown 
123 80 Ex2 Not found Unknown Unknown N/A 265.3 18 353 Unknown N/A Unknown 
124 20 Ex2 Not found Unknown Unknown N/A 0.5 13.4 11.9 371 Unknown 47, XY, +21c Unknown 
125 39 Ex2 Not found Unknown Unknown N/A 12.5 16.1 193 Unknown N/A Unknown 
126 35 Ex 3.1 Del 16 4871 Frameshift and introduction of stop codon 33.5 9.1 695 PTC 2 N/A Unknown 
127 30 Ex 3.1 Dup 4778 Frameshift and introduction of stop codon 21.7 13 24 PTC 2 N/A Unknown 
128 13.5 N/D Not found Unknown Unknown N/A 0.25 8.91 18.4 46 Unknown 47,XX,+21c[17] CCR 
129 N/D Not found Unknown Unknown N/A 10.6 18.9 65 Unknown N/A CCR 
130 N/D Not found Unknown Unknown N/A 0.75 8.3 8.1 100 Unknown N/A Unknown 
131 N/D Not found Unknown Unknown N/A 22.3 21 60 Unknown N/A Unknown 
132 N/D Not found Unknown Unknown N/A 0.5 15.2 20.8 276 Unknown N/A Unknown 
133 12 N/D Not found Unknown Unknown N/A N/A N/A N/A Unknown N/A Unknown 
134 N/D Not found Unknown Unknown N/A 4.15 13 100 Unknown N/A Unknown 

WBC indicates white blood cell; Hb, hemoglobin; Plt, platelet; Point, point substitution; CCR, complete clinical remission; Dup, duplication of nucleotides; Ins, insertion of nucleotides; Del, deletion of nucleotides; N/A, not available; and N/D, not done.

*

Exon number containing the mutation as defined by WAVE analysis.

Nucleotide 0 is the first nucleotide of GATA1 exon 1 including exons and introns. NCBI reference NT_079573.4 (Homo sapiens chromosome X genomic contig, starting position 11496706) was used.

Patients with TMD progressed to ML-DS.

Table 2

Summary of GATA1 mutations and patient characteristics for ML-DS samples

Patient no. Blast count, % of total nucleated count WAVE* Mutation Size, bp Position of mutation† Predicted effect of mutation on RNA Age at diagnosis, mo WBC count, ×109/L Hb level, g/dL Plt count, ×109/L Kanezaki et al15  class Karyotype Outcome 
70 Ex2 Del 56 4750 Frameshift and introduction of stop codon 24 12.6 5.7 22 PTC 1-3′ 47, XY, Del(7)(q32), +8, +21c CCR 
15 Ex2 Dup 13 4709 Frameshift and introduction of stop codon N/A 5.3 7.6 22 PTC 1-5′ 47, XX, +21c CCR 
84 Ex2 Ins 11 4724 Frameshift and introduction of stop codon 23 27.7 8.5 19 PTC 1-3′ 47, XX, +21c CCR 
4‡ 90 Ex2 Dup 20 4722 Frameshift and introduction of stop codon 36 8.5 11.3 20 PTC 1-3′ 56, XX, +2, +6, +8, +13, +13, +14, +14, −16, del(17)(q?22−24), +19, +del(19) (p13), +21, +21c[7], CCR 
20 Ex2 Point 4549 Loss of start codon 20 3.16 11 36 1st Met 48, XY, +8, +21 CCR 
N/A Ex2 Del 13 4753 Frameshift and introduction of stop codon 40 8.9 27 PTC 1-3′ 48, XY, der(1)add(1) (p32)dup(1)(q21q42), del(3)(q21q26),I(7) (q10),del(16)(q22q24), +21 Died 
14 Ex2 Del 10 4767 Frameshift and introduction of stop codon 21 27.8 5.4 85 PTC 1-3′ 47, XY, +21c Died 
N/A Ex2 Point 4548 Loss of splice acceptor site 24 4.1 2.1 Splice 47XX,i(7)(q10),−16, +21c, +mar[7] CCR 
N/A Ex2 Dup 20 4728 Frameshift and introduction of stop codon 1.2 11.9 7.5 18 PTC 1-3′ 47,XX, add(1)(q24), del(5)(p13), +21c CCR 
10 50 Ex2 Point 4768 Loss of splice acceptor site 31 2.8 9.7 21 Splice 48, XX, Add(8)(p23), +11, +21c[16]/ 47, XX, +21c [19] CCR 
11 N/A Ex2 Del 29 4669 Frameshift and introduction of stop codon 14 13 112 PTC 1-5′ 47, XY, +21, +8 CCR 
12 N/A Ex2 Point 4768 Loss of splice acceptor site 24 33.4 2.9 3.6 Splice 46,XX, der(7)add(7)(p?), −13,+21c[9]/47, XX,+21c Died 
13 60 Ex2 Point 4768 Loss of splice acceptor site 60 17.4 11 32 Splice 55∼57, XX, del(1)(q21;q25), +2, +3?adk1(5)(q33), i(7)(q10), +8, der(12)(t1;12)(q21;p13), +13, +14, +18, +19, +21, +22 Died 
14 20 Ex2 Point 4768 Loss of splice acceptor site 26 11.2 9.7 28 Splice 47. XX. +21c[36] Unknown 
15 78 Ex2 Ins 4736 Frameshift and introduction of stop codon 25 42.8 8.2 33 PTC 1-3′ 47, XY, dup(X) (p11.2,p21), +21c [2], 47, idem, der (4), t(2;4)(q31;q31), ?del(6)(q2?5; q2?7), +21c [5], 47, XY, +21c[3] CCR 
16 37 Ex2 Ins 4693 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-5′ N/A Relapsed 
17 16 Ex2 Point 4597 Nonsynonymous change in amino acid sequence introducing a stop codon 31 1.7 8.7 14 PTC 1-5′ 48, XX, + 8, +21c Unknown 
18 N/A Ex2 Ins 4632 Frameshift and introduction of stop codon 17 29 11.5 PTC 1-5′ 47 XX, +21c Died 
19 36 Ex2 Del 35 4684 Frameshift and introduction of stop codon 36 9.8 24 PTC 1-5′ 48, XX, +8, +21 CCR 
20 93 Ex2 Ins 4769 Loss of splice acceptor site N/A 200.3 4.7 23 Splice 47, XX, t(3;17) (q25;q25), del(6)(q13;q22), +21c[19] CCR 
21 20 Ex2 Del 4705 Frameshift and introduction of stop codon 24 5.4 3.7 PTC 1-5′ 47, XY, +21, +8 Died 
22‡ 23 Ex2 Dup 4717 Frameshift and introduction of stop codon 32 8.7 22 PTC 1-5′ 47, XY, t(5;11) (p15;q13), add(9)(q34), +21c[6]/ 47, XY, +21c[14] CCR 
23 21 Ex2 Del 4638 Frameshift and introduction of stop codon 11 6.4 15.1 5.1 PTC 1-5′ 47, XX, del (6), del (21), +21 Died 
24 44 Ex2 Del 23 4705 Frameshift and introduction of stop codon 13 8.3 N/A N/A PTC 1-5′ 48, XX, del (6) (q?13q?21) +21c, +21 [5] CCR 
25 50 Ex2 Del 4638 Frameshift and introduction of stop codon 12 7.3 N/A N/A PTC 1-5′ 50, XY, +10, +21c, +21, +22 [6] CCR 
26‡ 16 N/D Dup 10 4710 Frameshift and introduction of stop codon 11 3.5 11.9 30 PTC 1-5′ 47∼49,XY, +?19,+21c, +21[cp7]/47, XY,+21c[5] CCR 
27 20 N/D Dup 15 4715 Frameshift and introduction of stop codon 15 6.4 7.6 60 PTC 1-5′ 47,XY,+21c 2/47, idem,t(4;15), (q?21;q?21), del(7)(q?31q?33) Unknown 
28 8.5 N/D Del 4698 Frameshift and introduction of stop codon 13.4 70 PTC 1-5′ N/A Unknown 
29 31 N/D Del 4704 Frameshift and introduction of stop codon 13 3.9 11.2 47 PTC 1-5′ 47,XX,der(5)t(1;5) (q12;p14)?del(1) (q23q25),+21c[4]/48, XX,+8,der (12)t(1;12)(q12;p12), +21c[2]/47, XX,+21c[9] Unknown 
30 93 N/D Del 4638 Frameshift and introduction of stop codon 24 26.4 9.1 147 PTC 1-5′ 47,XY,+21c[25] Unknown 
31 39 N/D Del 4638 Frameshift and introduction of stop codon 27 8.5 8.5 26 PTC 1-5′ N/A Unknown 
32‡ 4.5 N/D Del 4604 Frameshift and introduction of stop codon 13 7.7 107 PTC 1-5′ N/A Unknown 
33 19.5 N/D Ins + Dup 2 + 14 4722 Frameshift and introduction of stop codon 21 26.4 7.2 151 PTC 1-3′ N/A Unknown 
34 50.5 N/D Ins 4768 Frameshift and introduction of stop codon 28 1.2 109 Splice 47∼48,XX,?add (5)(p1?5),−8, +21,+21c, +mar1, +mar2[cp8]/47, XX,+21c Unknown 
35 10.5 N/D Point 4768 Loss of splice acceptor site 35 3.4 10.4 17 Splice N/A Unknown 
36 36 N/D Dup 4734 Frameshift and introduction of stop codon 14 12.4 7.2 144 PTC 1-3′ 47,XX,+21c Unknown 
37 34 N/D Del 22 4704 Frameshift and introduction of stop codon 22 15.1 9.6 257 PTC 1-5′ 48,XY,+8,+21c Unknown 
38 55 N/D Del + Ins 31 + 6 4783 Loss of splice donor site 31 1.9 6.9 22 Splice N/A Unknown 
39 35 N/D Dup 4737 Frameshift and introduction of stop codon 31 4.2 9.2 20 PTC 1-3′ N/A Unknown 
40 38 N/D Del + ins 24 + 4 4683 Frameshift and introduction of stop codon 31 6.1 9.8 16 PTC 1-5′ N/A Unknown 
41 43 N/D Dup 21 4736 Frameshift and introduction of stop codon 13 4.9 6.7 38 PTC 1-3′ N/A Unknown 
42 15 N/D Dup 4668 Frameshift and introduction of stop codon 20 8.1 12.7 219 PTC 1-5′ 47,XX,+21c Unknown 
43 16 N/D Dup 4717 Frameshift and introduction of stop codon 36 160 1.5 PTC 1-5′ 47,XY,+21c,[3]/47, idem,t(1;8)(q32;q22) [2]/48,idem,t(1;8) (q32;q22),+der(8)t (1;8)(q32;q22) [10}] Unknown 
44 24 N/D Ins 4708 Frameshift and introduction of stop codon 10 6.88 9.7 35 PTC 1-5′ N/A Unknown 
45 11 N/D Point 4956 Synonymous mutation 16 2.5 8.1 101 Unknown N/A Unknown 
46 28 N/D Ins 4736 Frameshift and introduction of stop codon 31 12.9 3.1 76 PTC 1-3′ N/A Unknown 
47 14 N/D Point 4535 Unknown 31 22 8.5 37 Unknown 48, XY, +14, +21 Unknown 
48 13 N/D Del 4546 Loss of Start codon 19 6.2 10.6 131 1st Met 48,XY,i(7)(q10), t(11;11)(p15;q13), +21,+21[13]/47,XY,+21[2] Unknown 
49 15 N/D Point 4768 Loss of splice acceptor site 12 12.1 45 Splice N/A Unknown 
50 15 N/D Del 4702 Frameshift and introduction of stop codon 20 2.74 10.7 110 PTC 1-5′ 50,XX,+8, +14,+21,+21c, inc[5]/47, XX,+21c[7] Unknown 
51‡ 16 N/D Point 4552 Nonsynonymous change in amino acid sequence 12 3.4 11.1 27 Unknown 47,XY,?der(11) (q24∼25), +21c[2]/47, XY,+21c[8] Unknown 
52 N/D Del + ins 2 + 1 4619 Frameshift and introduction of stop codon 15 3.5 9.2 47 PTC 1-5′ 46,XY,i(7) (q10),der(21;21) (q10;q10)c, +21[6]/46,XY,der(21;21) (q10;q10)c, +21[5] Unknown 
53 12 N/D Del + ins 3 + 1 4654 Frameshift and introduction of stop codon 2.9 3.3 17 PTC 1-5′ 48,XX,der(1) (p?q?),+21c, +mar[14]/47,XX,+21c[4] Unknown 
54 10.5 N/D Dup 20 4727 Frameshift and introduction of stop codon 16 4.8 12 59 PTC 1-3′ N/A Unknown 
55 17 N/D Not Found Unknown Unknown Unknown 21 5.9 11.4 42 Unknown 47,XX,+21c Unknown 
56 N/A N/D Dup 16 4734 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-3′ N/A Unknown 
57 23 N/D Point 4767 Loss of splice acceptor site 26 5.1 13.3 11 Splice 48∼50,XY, +8,+?14, +21,+21c[cp6]/47,XY,+21c[4] Unknown 
58 21 N/D Point + Del 1 + 1 4637 + 4639 Frameshift and introduction of stop codon 37 6.3 11.4 13.7 Unknown 47,XX,?del(2)(q35), del(7)(p12),+21c[7]/47,XX,+21c[5] Unknown 
59 13 N/D Del 4698 Frameshift and introduction of stop codon 11 6.3 10.3 84 PTC 1-5′ 47,XY,+21c[9]; 48,XY,del(5) (p11p14),r(7) (p22q11), +8,+21c[7] Unknown 
60 35 N/D Del 4548 Unknown 11 16 38 Unknown N/A Unknown 
61 N/D Del 4642 Frameshift and introduction of stop codon 15 1.8 9.3 43 PTC 1-5′ 47,XY,+21[1] Unknown 
62 N/D Not Found Unknown Unknown Unknown 19 2.6 5.8 109 Unknown N/A Unknown 
63 20.5 N/D Del + ins 4 + 2 4656 Frameshift and introduction of stop codon 11 3.7 10.7 130 PTC 1-5′ 48,XX,+8,+21c[3]/47,XX,+21c[8] Unknown 
64 2.5 N/D Del 4656 Frameshift and introduction of stop codon 32 4.8 7.4 17 PTC 1-5′ 47,XX,+21c[6] Unknown 
65 46 N/D Dup 16 4719 Frameshift and introduction of stop codon 20 5.6 12 19 PTC 1-3′ N/A Unknown 
66 19.5 N/D Point 4767 Loss of splice acceptor site 32 5.46 13.6 52 Splice N/A Unknown 
67 N/D Del 4737 Frameshift and introduction of stop codon 26 3.3 7.1 69 PTC 1-3′ N/A Unknown 
68 16 N/D Del 4638 Frameshift and introduction of stop codon 35 2.8 9.5 20 PTC 1-5′ 48,XX,+8,+21c Unknown 
69 23 N/D Del 4638 Frameshift and introduction of stop codon 13 3.2 25 PTC 1-5′ 47,XY,+21c[10] Unknown 
70 76 N/D Del 4638 Frameshift and introduction of stop codon 17 23 6.5 28 PTC 1-5′ 46,XY,t(4;13) (q33∼34;q14), der(5)t(5;7) (p14∼15;q12), −7,+21c[cp11] Unknown 
71 3.5 N/D Del 4656 Frameshift and introduction of stop codon 13 3.3 11.4 46 PTC 1-5′ 47,XX,+21c[17] Unknown 
72 N/A N/D Point 4637 Unknown N/A N/A N/A N/A Unknown N/A Unknown 
73 20 N/D Point 4713 Synonymous mutation 19 4.1 9.5 13 Unknown 48∼49,XY,+21, +21c,inc[cp2]/47,XY,+21c[12] Unknown 
74 42 N/D Not Found Unknown Unknown Unknown 24 5.4 10.3 21 Unknown 47,XY,+21c, ?der(22)(p12) [5]/47,XY, +21c[15] Unknown 
75‡ 25 N/D Dup 13 4744 Frameshift and introduction of stop codon 26 2.7 11.8 20 PTC 1-3′ 47,XY,+21.ish 21qter(D21 S1446 × 3) Unknown 
76 96 N/D Point 4737 Nonsynonymous change in amino acid sequence 22 124 5.9 21 Unknown 47,X,−X, del(6)(q13q16), +21c,+21 Unknown 
77 10.5 N/D Not Found Unknown Unknown Unknown 17 5.2 10.6 82 Unknown 47,XX,add(16) (p13),+21c[5]/47,XX,+21c[5] Unknown 
78 26 N/D Point 4956 Synonymous mutation 20 6.4 130 Unknown 48,XY,der(8)t (1;8)(q21; p22∼23), +11,+21c[7]/47,XY,+21c[8] Unknown 
79 11 N/D Point 4768 Loss of splice acceptor site 43 6.35 12 70 Splice N/A Unknown 
80 13 N/D Point 4633 Nonsynonymous change in amino acid sequence 16 3.3 10.5 84 Unknown N/A Unknown 
81‡ 26 N/D Del 4638 Frameshift and introduction of stop codon 11 3.4 10.2 28 PTC 1-5′ 47,XY,+21c[15] Unknown 
82 N/D Ins 4725 Frameshift and introduction of stop codon 14 10.2 70 PTC 1-3′ N/A Unknown 
83 N/D Dup 4714 Frameshift and introduction of stop codon 8.75 11.4 69 PTC 1-5′ N/A Unknown 
84 N/A N/D Dup 48 4771 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-3′ N/A Unknown 
85 17.5 N/D Del 4638 Frameshift and introduction of stop codon 11 4.7 9.3 83 PTC 1-5′ N/A Unknown 
86 45 N/D Dup 10 4723 Frameshift and introduction of stop codon 14 11.5 8.6 44 PTC 1-3′ N/A Unknown 
87 70.5 N/D Del 4638 Frameshift and introduction of stop codon 16 32.5 7.9 103 PTC 1-5′ 46,XY[22]. nuc ish 8q22(ETOx2), 21q22(AML1 × 2), 13q14(RB1 × 2), 13q34 × 2), 11q23(MLLx2)[100] Unknown 
88 31 N/D Del 4638 Frameshift and introduction of stop codon 29 11.4 13.2 57.4 PTC 1-5′ 46,XX,del (9)(q?13q33), der(19)t(1;19) (q31;p13), der(21;21) (q10;q10)?c Unknown 
89 7.5 N/D Del 22 4706 Frameshift and introduction of stop codon 17 7.8 11.1 27 PTC 1-5′ 48,XY,add(7) (p2?2),+8, del(13)(q1?4), +21c[7]/46, XY,−21,+21c or 46,XY[3]/47,XY,+21c[2] Unknown 
90 33 N/D Del + ins 2 + 6 4707 Frameshift and introduction of stop codon 4.6 7.5 PTC 1-5′ N/A Unknown 
91‡ 24 N/D Ins 4733 Frameshift and introduction of stop codon 11 7.1 57 PTC 1-3′ 48,X,add(Y) (q12),+11, +21c[8]/47,XY, +21c[7] Unknown 
92 28 N/D Dup 10 4719 Frameshift and introduction of stop codon 22 5.2 13.7 143 PTC 1-3′ N/A Unknown 
93 16 Ex2 Not Found N/A N/A N/A 1.5 10.4 13 Unknown 47, XY, +21c Died 
94 30 Ex2 Not Found N/A N/A N/A 15 14.47 4.1 38 Unknown 47, XX, t(3;3)(q21;q26), t(3;3)(p2?3;q13.3), +21c[13]/47, idem, der(3)t(3;3)add(3)(q29)[7] CCR 
95 20 Ex2 Not Found N/A N/A N/A 17 11 9.9 52 Unknown 47, XY, der(11)t (q23p15)t (1:11)(q23;q85), +21c CCR 
96 10 Ex2 Not Found N/A N/A N/A 11 N/A 8.6 86 Unknown 47, XY, del (7), +21 Unknown 
97 21 Ex2 Not Found N/A N/A N/A 33 4.9 15 Unknown 48, XY, +21, +21c CCR 
98 20 Ex2 Not Found N/A N/A N/A 15 15.2 9.9 18 Unknown 47 XX, ?dup(3) (p23p25), del(8) (p11.2p21), der(15)t(1:15) (q21;p11), del(16)(q22q24, del(20) CCR 
99 Ex2 Not Found N/A N/A N/A 23 5.1 7.2 49 Unknown N/A Unknown 
100 20 Ex2 Not Found N/A N/A N/A 38 N/A N/A N/A Unknown 46, XX, +21, −7 Unknown 
101 38 Ex2 Not Found N/A N/A N/A 21 11.9 7.3 34 Unknown 47, XX, +21c CCR 
102 10 Ex2 Not Found N/A N/A N/A 10 3.9 10.9 65 Unknown N/A Unknown 
103 20 Ex2 Not Found N/A N/A N/A 12 3.8 8.6 16 Unknown 47, XX, t(1;22)(q23;p13), +21c[9]/ 47, XX, +21c[4] CCR 
Patient no. Blast count, % of total nucleated count WAVE* Mutation Size, bp Position of mutation† Predicted effect of mutation on RNA Age at diagnosis, mo WBC count, ×109/L Hb level, g/dL Plt count, ×109/L Kanezaki et al15  class Karyotype Outcome 
70 Ex2 Del 56 4750 Frameshift and introduction of stop codon 24 12.6 5.7 22 PTC 1-3′ 47, XY, Del(7)(q32), +8, +21c CCR 
15 Ex2 Dup 13 4709 Frameshift and introduction of stop codon N/A 5.3 7.6 22 PTC 1-5′ 47, XX, +21c CCR 
84 Ex2 Ins 11 4724 Frameshift and introduction of stop codon 23 27.7 8.5 19 PTC 1-3′ 47, XX, +21c CCR 
4‡ 90 Ex2 Dup 20 4722 Frameshift and introduction of stop codon 36 8.5 11.3 20 PTC 1-3′ 56, XX, +2, +6, +8, +13, +13, +14, +14, −16, del(17)(q?22−24), +19, +del(19) (p13), +21, +21c[7], CCR 
20 Ex2 Point 4549 Loss of start codon 20 3.16 11 36 1st Met 48, XY, +8, +21 CCR 
N/A Ex2 Del 13 4753 Frameshift and introduction of stop codon 40 8.9 27 PTC 1-3′ 48, XY, der(1)add(1) (p32)dup(1)(q21q42), del(3)(q21q26),I(7) (q10),del(16)(q22q24), +21 Died 
14 Ex2 Del 10 4767 Frameshift and introduction of stop codon 21 27.8 5.4 85 PTC 1-3′ 47, XY, +21c Died 
N/A Ex2 Point 4548 Loss of splice acceptor site 24 4.1 2.1 Splice 47XX,i(7)(q10),−16, +21c, +mar[7] CCR 
N/A Ex2 Dup 20 4728 Frameshift and introduction of stop codon 1.2 11.9 7.5 18 PTC 1-3′ 47,XX, add(1)(q24), del(5)(p13), +21c CCR 
10 50 Ex2 Point 4768 Loss of splice acceptor site 31 2.8 9.7 21 Splice 48, XX, Add(8)(p23), +11, +21c[16]/ 47, XX, +21c [19] CCR 
11 N/A Ex2 Del 29 4669 Frameshift and introduction of stop codon 14 13 112 PTC 1-5′ 47, XY, +21, +8 CCR 
12 N/A Ex2 Point 4768 Loss of splice acceptor site 24 33.4 2.9 3.6 Splice 46,XX, der(7)add(7)(p?), −13,+21c[9]/47, XX,+21c Died 
13 60 Ex2 Point 4768 Loss of splice acceptor site 60 17.4 11 32 Splice 55∼57, XX, del(1)(q21;q25), +2, +3?adk1(5)(q33), i(7)(q10), +8, der(12)(t1;12)(q21;p13), +13, +14, +18, +19, +21, +22 Died 
14 20 Ex2 Point 4768 Loss of splice acceptor site 26 11.2 9.7 28 Splice 47. XX. +21c[36] Unknown 
15 78 Ex2 Ins 4736 Frameshift and introduction of stop codon 25 42.8 8.2 33 PTC 1-3′ 47, XY, dup(X) (p11.2,p21), +21c [2], 47, idem, der (4), t(2;4)(q31;q31), ?del(6)(q2?5; q2?7), +21c [5], 47, XY, +21c[3] CCR 
16 37 Ex2 Ins 4693 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-5′ N/A Relapsed 
17 16 Ex2 Point 4597 Nonsynonymous change in amino acid sequence introducing a stop codon 31 1.7 8.7 14 PTC 1-5′ 48, XX, + 8, +21c Unknown 
18 N/A Ex2 Ins 4632 Frameshift and introduction of stop codon 17 29 11.5 PTC 1-5′ 47 XX, +21c Died 
19 36 Ex2 Del 35 4684 Frameshift and introduction of stop codon 36 9.8 24 PTC 1-5′ 48, XX, +8, +21 CCR 
20 93 Ex2 Ins 4769 Loss of splice acceptor site N/A 200.3 4.7 23 Splice 47, XX, t(3;17) (q25;q25), del(6)(q13;q22), +21c[19] CCR 
21 20 Ex2 Del 4705 Frameshift and introduction of stop codon 24 5.4 3.7 PTC 1-5′ 47, XY, +21, +8 Died 
22‡ 23 Ex2 Dup 4717 Frameshift and introduction of stop codon 32 8.7 22 PTC 1-5′ 47, XY, t(5;11) (p15;q13), add(9)(q34), +21c[6]/ 47, XY, +21c[14] CCR 
23 21 Ex2 Del 4638 Frameshift and introduction of stop codon 11 6.4 15.1 5.1 PTC 1-5′ 47, XX, del (6), del (21), +21 Died 
24 44 Ex2 Del 23 4705 Frameshift and introduction of stop codon 13 8.3 N/A N/A PTC 1-5′ 48, XX, del (6) (q?13q?21) +21c, +21 [5] CCR 
25 50 Ex2 Del 4638 Frameshift and introduction of stop codon 12 7.3 N/A N/A PTC 1-5′ 50, XY, +10, +21c, +21, +22 [6] CCR 
26‡ 16 N/D Dup 10 4710 Frameshift and introduction of stop codon 11 3.5 11.9 30 PTC 1-5′ 47∼49,XY, +?19,+21c, +21[cp7]/47, XY,+21c[5] CCR 
27 20 N/D Dup 15 4715 Frameshift and introduction of stop codon 15 6.4 7.6 60 PTC 1-5′ 47,XY,+21c 2/47, idem,t(4;15), (q?21;q?21), del(7)(q?31q?33) Unknown 
28 8.5 N/D Del 4698 Frameshift and introduction of stop codon 13.4 70 PTC 1-5′ N/A Unknown 
29 31 N/D Del 4704 Frameshift and introduction of stop codon 13 3.9 11.2 47 PTC 1-5′ 47,XX,der(5)t(1;5) (q12;p14)?del(1) (q23q25),+21c[4]/48, XX,+8,der (12)t(1;12)(q12;p12), +21c[2]/47, XX,+21c[9] Unknown 
30 93 N/D Del 4638 Frameshift and introduction of stop codon 24 26.4 9.1 147 PTC 1-5′ 47,XY,+21c[25] Unknown 
31 39 N/D Del 4638 Frameshift and introduction of stop codon 27 8.5 8.5 26 PTC 1-5′ N/A Unknown 
32‡ 4.5 N/D Del 4604 Frameshift and introduction of stop codon 13 7.7 107 PTC 1-5′ N/A Unknown 
33 19.5 N/D Ins + Dup 2 + 14 4722 Frameshift and introduction of stop codon 21 26.4 7.2 151 PTC 1-3′ N/A Unknown 
34 50.5 N/D Ins 4768 Frameshift and introduction of stop codon 28 1.2 109 Splice 47∼48,XX,?add (5)(p1?5),−8, +21,+21c, +mar1, +mar2[cp8]/47, XX,+21c Unknown 
35 10.5 N/D Point 4768 Loss of splice acceptor site 35 3.4 10.4 17 Splice N/A Unknown 
36 36 N/D Dup 4734 Frameshift and introduction of stop codon 14 12.4 7.2 144 PTC 1-3′ 47,XX,+21c Unknown 
37 34 N/D Del 22 4704 Frameshift and introduction of stop codon 22 15.1 9.6 257 PTC 1-5′ 48,XY,+8,+21c Unknown 
38 55 N/D Del + Ins 31 + 6 4783 Loss of splice donor site 31 1.9 6.9 22 Splice N/A Unknown 
39 35 N/D Dup 4737 Frameshift and introduction of stop codon 31 4.2 9.2 20 PTC 1-3′ N/A Unknown 
40 38 N/D Del + ins 24 + 4 4683 Frameshift and introduction of stop codon 31 6.1 9.8 16 PTC 1-5′ N/A Unknown 
41 43 N/D Dup 21 4736 Frameshift and introduction of stop codon 13 4.9 6.7 38 PTC 1-3′ N/A Unknown 
42 15 N/D Dup 4668 Frameshift and introduction of stop codon 20 8.1 12.7 219 PTC 1-5′ 47,XX,+21c Unknown 
43 16 N/D Dup 4717 Frameshift and introduction of stop codon 36 160 1.5 PTC 1-5′ 47,XY,+21c,[3]/47, idem,t(1;8)(q32;q22) [2]/48,idem,t(1;8) (q32;q22),+der(8)t (1;8)(q32;q22) [10}] Unknown 
44 24 N/D Ins 4708 Frameshift and introduction of stop codon 10 6.88 9.7 35 PTC 1-5′ N/A Unknown 
45 11 N/D Point 4956 Synonymous mutation 16 2.5 8.1 101 Unknown N/A Unknown 
46 28 N/D Ins 4736 Frameshift and introduction of stop codon 31 12.9 3.1 76 PTC 1-3′ N/A Unknown 
47 14 N/D Point 4535 Unknown 31 22 8.5 37 Unknown 48, XY, +14, +21 Unknown 
48 13 N/D Del 4546 Loss of Start codon 19 6.2 10.6 131 1st Met 48,XY,i(7)(q10), t(11;11)(p15;q13), +21,+21[13]/47,XY,+21[2] Unknown 
49 15 N/D Point 4768 Loss of splice acceptor site 12 12.1 45 Splice N/A Unknown 
50 15 N/D Del 4702 Frameshift and introduction of stop codon 20 2.74 10.7 110 PTC 1-5′ 50,XX,+8, +14,+21,+21c, inc[5]/47, XX,+21c[7] Unknown 
51‡ 16 N/D Point 4552 Nonsynonymous change in amino acid sequence 12 3.4 11.1 27 Unknown 47,XY,?der(11) (q24∼25), +21c[2]/47, XY,+21c[8] Unknown 
52 N/D Del + ins 2 + 1 4619 Frameshift and introduction of stop codon 15 3.5 9.2 47 PTC 1-5′ 46,XY,i(7) (q10),der(21;21) (q10;q10)c, +21[6]/46,XY,der(21;21) (q10;q10)c, +21[5] Unknown 
53 12 N/D Del + ins 3 + 1 4654 Frameshift and introduction of stop codon 2.9 3.3 17 PTC 1-5′ 48,XX,der(1) (p?q?),+21c, +mar[14]/47,XX,+21c[4] Unknown 
54 10.5 N/D Dup 20 4727 Frameshift and introduction of stop codon 16 4.8 12 59 PTC 1-3′ N/A Unknown 
55 17 N/D Not Found Unknown Unknown Unknown 21 5.9 11.4 42 Unknown 47,XX,+21c Unknown 
56 N/A N/D Dup 16 4734 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-3′ N/A Unknown 
57 23 N/D Point 4767 Loss of splice acceptor site 26 5.1 13.3 11 Splice 48∼50,XY, +8,+?14, +21,+21c[cp6]/47,XY,+21c[4] Unknown 
58 21 N/D Point + Del 1 + 1 4637 + 4639 Frameshift and introduction of stop codon 37 6.3 11.4 13.7 Unknown 47,XX,?del(2)(q35), del(7)(p12),+21c[7]/47,XX,+21c[5] Unknown 
59 13 N/D Del 4698 Frameshift and introduction of stop codon 11 6.3 10.3 84 PTC 1-5′ 47,XY,+21c[9]; 48,XY,del(5) (p11p14),r(7) (p22q11), +8,+21c[7] Unknown 
60 35 N/D Del 4548 Unknown 11 16 38 Unknown N/A Unknown 
61 N/D Del 4642 Frameshift and introduction of stop codon 15 1.8 9.3 43 PTC 1-5′ 47,XY,+21[1] Unknown 
62 N/D Not Found Unknown Unknown Unknown 19 2.6 5.8 109 Unknown N/A Unknown 
63 20.5 N/D Del + ins 4 + 2 4656 Frameshift and introduction of stop codon 11 3.7 10.7 130 PTC 1-5′ 48,XX,+8,+21c[3]/47,XX,+21c[8] Unknown 
64 2.5 N/D Del 4656 Frameshift and introduction of stop codon 32 4.8 7.4 17 PTC 1-5′ 47,XX,+21c[6] Unknown 
65 46 N/D Dup 16 4719 Frameshift and introduction of stop codon 20 5.6 12 19 PTC 1-3′ N/A Unknown 
66 19.5 N/D Point 4767 Loss of splice acceptor site 32 5.46 13.6 52 Splice N/A Unknown 
67 N/D Del 4737 Frameshift and introduction of stop codon 26 3.3 7.1 69 PTC 1-3′ N/A Unknown 
68 16 N/D Del 4638 Frameshift and introduction of stop codon 35 2.8 9.5 20 PTC 1-5′ 48,XX,+8,+21c Unknown 
69 23 N/D Del 4638 Frameshift and introduction of stop codon 13 3.2 25 PTC 1-5′ 47,XY,+21c[10] Unknown 
70 76 N/D Del 4638 Frameshift and introduction of stop codon 17 23 6.5 28 PTC 1-5′ 46,XY,t(4;13) (q33∼34;q14), der(5)t(5;7) (p14∼15;q12), −7,+21c[cp11] Unknown 
71 3.5 N/D Del 4656 Frameshift and introduction of stop codon 13 3.3 11.4 46 PTC 1-5′ 47,XX,+21c[17] Unknown 
72 N/A N/D Point 4637 Unknown N/A N/A N/A N/A Unknown N/A Unknown 
73 20 N/D Point 4713 Synonymous mutation 19 4.1 9.5 13 Unknown 48∼49,XY,+21, +21c,inc[cp2]/47,XY,+21c[12] Unknown 
74 42 N/D Not Found Unknown Unknown Unknown 24 5.4 10.3 21 Unknown 47,XY,+21c, ?der(22)(p12) [5]/47,XY, +21c[15] Unknown 
75‡ 25 N/D Dup 13 4744 Frameshift and introduction of stop codon 26 2.7 11.8 20 PTC 1-3′ 47,XY,+21.ish 21qter(D21 S1446 × 3) Unknown 
76 96 N/D Point 4737 Nonsynonymous change in amino acid sequence 22 124 5.9 21 Unknown 47,X,−X, del(6)(q13q16), +21c,+21 Unknown 
77 10.5 N/D Not Found Unknown Unknown Unknown 17 5.2 10.6 82 Unknown 47,XX,add(16) (p13),+21c[5]/47,XX,+21c[5] Unknown 
78 26 N/D Point 4956 Synonymous mutation 20 6.4 130 Unknown 48,XY,der(8)t (1;8)(q21; p22∼23), +11,+21c[7]/47,XY,+21c[8] Unknown 
79 11 N/D Point 4768 Loss of splice acceptor site 43 6.35 12 70 Splice N/A Unknown 
80 13 N/D Point 4633 Nonsynonymous change in amino acid sequence 16 3.3 10.5 84 Unknown N/A Unknown 
81‡ 26 N/D Del 4638 Frameshift and introduction of stop codon 11 3.4 10.2 28 PTC 1-5′ 47,XY,+21c[15] Unknown 
82 N/D Ins 4725 Frameshift and introduction of stop codon 14 10.2 70 PTC 1-3′ N/A Unknown 
83 N/D Dup 4714 Frameshift and introduction of stop codon 8.75 11.4 69 PTC 1-5′ N/A Unknown 
84 N/A N/D Dup 48 4771 Frameshift and introduction of stop codon N/A N/A N/A N/A PTC 1-3′ N/A Unknown 
85 17.5 N/D Del 4638 Frameshift and introduction of stop codon 11 4.7 9.3 83 PTC 1-5′ N/A Unknown 
86 45 N/D Dup 10 4723 Frameshift and introduction of stop codon 14 11.5 8.6 44 PTC 1-3′ N/A Unknown 
87 70.5 N/D Del 4638 Frameshift and introduction of stop codon 16 32.5 7.9 103 PTC 1-5′ 46,XY[22]. nuc ish 8q22(ETOx2), 21q22(AML1 × 2), 13q14(RB1 × 2), 13q34 × 2), 11q23(MLLx2)[100] Unknown 
88 31 N/D Del 4638 Frameshift and introduction of stop codon 29 11.4 13.2 57.4 PTC 1-5′ 46,XX,del (9)(q?13q33), der(19)t(1;19) (q31;p13), der(21;21) (q10;q10)?c Unknown 
89 7.5 N/D Del 22 4706 Frameshift and introduction of stop codon 17 7.8 11.1 27 PTC 1-5′ 48,XY,add(7) (p2?2),+8, del(13)(q1?4), +21c[7]/46, XY,−21,+21c or 46,XY[3]/47,XY,+21c[2] Unknown 
90 33 N/D Del + ins 2 + 6 4707 Frameshift and introduction of stop codon 4.6 7.5 PTC 1-5′ N/A Unknown 
91‡ 24 N/D Ins 4733 Frameshift and introduction of stop codon 11 7.1 57 PTC 1-3′ 48,X,add(Y) (q12),+11, +21c[8]/47,XY, +21c[7] Unknown 
92 28 N/D Dup 10 4719 Frameshift and introduction of stop codon 22 5.2 13.7 143 PTC 1-3′ N/A Unknown 
93 16 Ex2 Not Found N/A N/A N/A 1.5 10.4 13 Unknown 47, XY, +21c Died 
94 30 Ex2 Not Found N/A N/A N/A 15 14.47 4.1 38 Unknown 47, XX, t(3;3)(q21;q26), t(3;3)(p2?3;q13.3), +21c[13]/47, idem, der(3)t(3;3)add(3)(q29)[7] CCR 
95 20 Ex2 Not Found N/A N/A N/A 17 11 9.9 52 Unknown 47, XY, der(11)t (q23p15)t (1:11)(q23;q85), +21c CCR 
96 10 Ex2 Not Found N/A N/A N/A 11 N/A 8.6 86 Unknown 47, XY, del (7), +21 Unknown 
97 21 Ex2 Not Found N/A N/A N/A 33 4.9 15 Unknown 48, XY, +21, +21c CCR 
98 20 Ex2 Not Found N/A N/A N/A 15 15.2 9.9 18 Unknown 47 XX, ?dup(3) (p23p25), del(8) (p11.2p21), der(15)t(1:15) (q21;p11), del(16)(q22q24, del(20) CCR 
99 Ex2 Not Found N/A N/A N/A 23 5.1 7.2 49 Unknown N/A Unknown 
100 20 Ex2 Not Found N/A N/A N/A 38 N/A N/A N/A Unknown 46, XX, +21, −7 Unknown 
101 38 Ex2 Not Found N/A N/A N/A 21 11.9 7.3 34 Unknown 47, XX, +21c CCR 
102 10 Ex2 Not Found N/A N/A N/A 10 3.9 10.9 65 Unknown N/A Unknown 
103 20 Ex2 Not Found N/A N/A N/A 12 3.8 8.6 16 Unknown 47, XX, t(1;22)(q23;p13), +21c[9]/ 47, XX, +21c[4] CCR 

WBC indicates white blood cell; Hb, hemoglobin; Plt, platelet; Del, deletion of nucleotides; CCR, complete clinical remission; Dup, duplication of nucleotides; N/A, not available; Ins, insertion of nucleotides; Point, point substitution; and N/D, not done.

*

Exon number containing the mutation as defined by WAVE analysis.

Nucleotide 0 is the first nucleotide of GATA1 exon 1 including exons and introns. NCBI reference NT_079573.4 (Homo sapiens chromosome X genomic contig, starting position 11496706) was used.

Patients with TMD progressed to ML-DS.

GATA1 mutations were analyzed by WAVE and direct sequencing of PCR products. GATA1 sequence mutations were determined in 118 of 134 patients with TMD (88.1%) and in 88 of 103 patients with ML-DS (85.4%). Mutations were detected by WAVE in a further 9 and 11 patients, respectively (Tables 12). The main reason for failure to detect mutations was low blast count. Alternative techniques such as high-resolution melt analysis or nested PCR may be able to detect mutations in some cases. The lower limit of blasts that allowed successful mutation detection was 0.5% (supplemental Methods). However, in one patient (blast count, 42%) failure to detect mutation suggests an uncommon mutation involving sequence outside the genomic area, spanning the PCR, or a deletion inside this area, affecting the primer annealing site.

Relative positions of sequence mutations are shown in Figure 1A. Insertion/deletion/duplications comprised 78% of mutations in both TMD and ML-DS (Figure 1B), consistent with previous reports.12  Point mutations were detected in 21% and 22% of TMD and ML-DS samples, respectively. Substitutions were rare, uniquely detected in 1% of patients with TMD. Therefore, there is little difference in mutational spectrum between TMD and ML-DS. Thirteen patients with TMD are known to have progressed to ML-DS. In these samples the spectrum of mutations was similar to the TMD group that did not progress to ML-DS (insertion/deletion/duplications, 77%; point mutations, 23%). Eight patients had paired TMD and follow-up ML-DS samples. The same mutation was present in both TMD and ML-DS for 7 of 8 samples, similar to previous observations.12,14 

Figure 1

Position and types of GATA1 sequence mutations found in TMD and ML-DS samples. (A) A schematic diagram of GATA1 showing the positions and types of the sequence mutations found in TMD and ML-DS samples. Each arrow represents a different patient. (B) Diagram showing the mutational spectrum of patients with TMD and with ML-DS and (C) the effect that these mutations have on the sequence of GATA1.

Figure 1

Position and types of GATA1 sequence mutations found in TMD and ML-DS samples. (A) A schematic diagram of GATA1 showing the positions and types of the sequence mutations found in TMD and ML-DS samples. Each arrow represents a different patient. (B) Diagram showing the mutational spectrum of patients with TMD and with ML-DS and (C) the effect that these mutations have on the sequence of GATA1.

Predicted consequence of mutations was similar for both TMD and ML-DS (Figure 1C). Most mutations inserted a premature termination codon (PTC) either by introducing a stop codon or frameshift. Mutations affecting the splice site at GATA1 exon 2 exon/intron boundary were next most frequent. In some cases (3 TMD and 3 DS-ML) where a point mutation occurred in intronic sequence, predicted consequence was unclear. They may occur in splice site regulatory elements and may affect gene splicing.18  In 13 patients with TMD who progressed to ML-DS predicted consequences in the protein were similar.

Characterizing the sequence of GATA1 mutations provides an opportunity to develop patient mutation-specific quantitative PCR analysis to monitor resolution of TMD, persistence/re-emergence of GATA1 mutant clone leading to ML-DS, and therapy response in patients with ML-DS.13  Direct sequencing of DNA from blasts that underwent FACS was successful in identifying sequence mutation in 19 patients, whereas direct sequencing of DNA from unfractionated samples failed. In 20 cases it was necessary to subclone the GATA1 PCR product to pinpoint mutation sequence.

A previous study divided GATA1 mutations in TMD and ML-DS into 2 classes on the basis of levels of GATA1s protein expression.15  Mutations affecting gene splicing, the start codon (1st Met) or that introduced a PTC in the 3′ end of exon 2 (PTC 1-3′) resulted in high GATA1s protein levels; whereas a PTC in the 5′ end of exon 2 (PTC 1-5′) or in exon 3.1 (PTC-2) resulted in low GATA1s expression.15  Patients with TMD with mutations predicted to result in low GATA1s protein expression were more probable to develop ML-DS because this type of mutation had significantly higher incidence in ML-DS.15  We asked whether this was true for our sample cohort. No significant differences were found between numbers of samples in each of the mutation types when TMD and ML-DS were compared (supplementary Table 1; 1st Met, P = .7011; splice errors, P = .6741; PTC 1-3′, P = .3388; PTC 1-5′, P = .3021; PTC-2, P = .2667). Similarly, we found no significant increase in predicted GATA1s low-expressing mutations in ML-DS samples versus TMD (P = .5534; Tables 12). In 12 TMD samples that progressed to ML-DS whereby we obtained a sequence mutation predicted to affect protein expression, equal numbers were predicted to result in high and low GATA1s protein expression (ie, 6 of each). GATA1s cDNA qPCR for 36 TMD and 20 ML-DS patient samples showed no significant differences in GATA1s mRNA expression in patients with PTC 1-3′, Splice, PTC 1-5′, or unknown effect mutations (supplemental Figure 1).

Cytogenetic data were available for 31 patients with TMD and 68 patients with ML-DS (Tables 12). In neonates with TMD, no significant correlation was observed between not progressing to ML-DS and presenting with T21 as the only cytogenetic abnormality (P = .2533; supplemental Table 2). Therefore, karyotype does not predict patients who will progress to ML-DS.

In conclusion, simple techniques detect GATA1 mutations in most patients TMD and patients with ML-DS. It is not possible to predict progression of TMD to ML-DS on the basis of type of GATA1 mutation, at least in mainly white patients.

The online version of this article contains a data supplement.

The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked “advertisement” in accordance with 18 USC section 1734.

Acknowledgments

The authors thank Esther Edlundh-Rose for technical help processing patient samples.

This work was supported in Oxford by the Leukemia & Lymphoma Research Specialist Program (award 08030; K.A.A., P.V., and I.R.), NIHR Biomedical Research Centres funding scheme (P.V.), the Medical Research Council Disease Team Award (P.V.), the MRC Molecular Hematology Unit (P.V.), and the Imperial College London Comprehensive Biomedical Research Center (I.R.). This work was supported in Hannover by German Research Foundation (grant DFG RE2580/1-1) and by a grant from the Madeleine-Schickedanz Leukemia Foundation.

Authorship

Contribution: K.A.A. performed and analyzed experiments and wrote the manuscript; A.N. and K.R. designed, performed, and analyzed experiments and data; C.G. performed experiments, analyzed data, and compiled figures; K.B., C.v.N., and A.K. performed and analyzed experiments; E.M. analyzed data; and P.V., D.R., J-H.K., H.H., and I.R. designed experiments, analyzed data, and wrote or reviewed the manuscript.

Conflict-of-interest disclosure: The authors declare no competing financial interests.

A complete list of the participants of the International ML-DS Study Group can be found in the supplemental Appendix.

Correspondence: Paresh Vyas, Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom; e-mail: paresh.vyas@imm.ox.ac.uk; or Dirk Reinhardt, Department of Pediatric Hematology/Oncology, Medical School Hannover, Carl Neuberg Str 1, 30625 Hannover, Germany; e-mail: reinhardt.dirk@mh.hannover.de.

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Author notes

*

K.A.A., K.R., C.G., and A.N. contributed equally to the work and are joint first authors.