17.analysis of GATA1 Mutations in Down Syndrome Transient Myeloproliferative Disorder and Myeloid Leukemia PDF

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MYELOID NEOPLASIA
Brief report
Analysis of GATA1 mutations in Down syndrome transient myeloproliferative

disorder and myeloid leukemia
*Kate A. Alford,1 *Katarina Reinhardt,2 *Catherine Garnett,1 *Alice Norton,1 Katarina Bohmer,2 Christine von Neuhoff,2
Alexandra Kolenova,3 Emanuele Marchi,1 Jan-Henning Klusmann,2 Irene Roberts,4 Henrik Hasle,5 Dirk Reinhardt,2 and
Paresh Vyas,1 on behalf of the International Myeloid LeukemiaDown Syndrome Study Group
1Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom; 2Pediatric
Hematology and Oncology, Hannover Medical School, Hannover, Germany; 3Department of Pediatric Oncology, Comenius University Medical School and
University Childrens Hospital, Bratislava, Slovakia; 4Department of Haematology, Imperial College London, London, United Kingdom; and 5Nordic Society of
Pediatric Hematology and Oncology, Department of Pediatrics, Aarhus University Hospital, Skejby, Denmark
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 simi-
lar 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 sig-
nificant 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. (Blood.
2011;118(8):2222-2238)
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
Submitted March 17, 2011; accepted June 13, 2011. Prepublished online as
Blood First Edition paper, June 29, 2011; DOI 10.1182/blood-2011-03-342774.
*K.A.A., K.R., C.G., and A.N. contributed equally to the work and are joint first
authors.
2222
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-Munster 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
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.
2011 by The American Society of Hematology
BLOOD, 25 AUGUST 2011 VOLUME 118, NUMBER 8
(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).
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 1-2). 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
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
GATA1 MUTATIONS IN TMD AND ML-DS
2223
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 1-2). 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 1-2). 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.
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.
21
60
35
82
75
12
13
14
15
16
17
18
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ins
Point
Point
Point
Ins
Del
Del
Dup
Ins
Del
Point
Dup
Dup
Point
Dup
Del
Dup
Dup
Ins
Point
Dup
Point
16
26
22
16
17
22
19
101
20
No. of
nucleotides
changed, bp
4709
4766
4596
4550
4706
4458
4688
4723
4721
4607
4768
4719
4735
4583
4768
4638
4733
4653
4701
4767
4722
4549
Position of
mutation
of stop codon
Frameshift and introduction
Loss of splice acceptor site
introducing a stop codon
amino acid sequence
Nonsynonymous change in
Loss of start codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
Loss of start codon
Predicted effect of
mutation on RNA
N/A
N/A
1.1
0.71
0.57
0.71
0.28
0.28
0.57
0.57
1.3
Age at
diagnosis, mo
33.7
N/A
N/A
11.5
31.6
249
39.3
12
6.7
9.5
19.4
38.2
121
50
120
44.6
75.9
16
14
70
107
WBC count,
109/L
14.8
N/A
N/A
13.3
16.2
11.8
13.8
14
16.3
12.7
10.8
18.7
11.6
N/A
18.8
17.4
6.3
8.2
21.3
12.7
13.3
15
Hb level,
g/dL
35
N/A
N/A
72
44
122
335
88
59
54
52
132
35
N/A
505
45
113
122
41
461
352
140
Plt
count,
109/L
PTC 1-5
Splice
PTC 1-5
1st Met
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-3
PTC 1-5
Splice
PTC 1-3
PTC 1-3
PTC 1-5
Splice
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-5
Splice
PTC 1-3
1st Met
Kanezaki et al15
class
N/A
N/A
N/A
47, XY, 21c
N/A
47, XY, 21c
N/A
N/A
N/A
N/A
N/A
N/A
47, XY, 21
N/A
N/A
47, XX, 21c
47, XX, 21c
Unknown
Unknown
Died
CCR
CCR
CCR
CCR
Unknown
Died
Unknown
Unknown
CCR
cytopaenia
Thrombo-
ML-DS
CCR
CCR
CCR
CCR
CCR
47, XY, 21c

47, XY, 21c
CCR
47, XY, 21c
ML-DS
47, XX, 21c
Outcome
CCR
Karyotype
47, XY, 21c
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.
25
11
11
22
80
10
N/A
40
21
92
30
29
N/A
55
20
30
Ex2
Ex2
Ex2
Ex2
WAVE*
Mutation
type
ALFORD et al
19
40
35
66
79
Blast count, %
of total
nucleated count
2224
Patient
no.
Table 1. Summary of GATA1 mutations and patient characteristics for TMD samples
13.5
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Dup
Point
Del Ins
Del
Del Ins
Del
Point
Point
Ins
Dup
Dup
Del
Del
Point
Dup
Ins
Del
Dup
Del
Ins
Del
Substitution
36
12
21
14
10
40
132
No. of
nucleotides
changed, bp
4737
4734
4679
4680
4770 4774
4604
4767
4768
4707
4710
4740
4638
4697
4670
4732
4643
4684
4717
4662
4661
4671
4744
Position of
mutation
of stop codon
Unknown
of stop codon
of stop codon
Loss of splice donor site
of stop codon
Synonymous mutation
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
Predicted effect of
mutation on RNA
0.5
0.25
1.25
0.25
0.25
gestation
28 weeks
2.7
Age at
diagnosis, mo
3.2
16.9
172.8
232
9.1
1193.3
19.2
55.6
55.6
46.6
9.8
198.2
40
21.7
33.5
N/A
200
101
39.6
124.9
58
20
WBC count,
109/L
11.1
14.9
16.2
15.6
9.6
14.7
6.7
17.3
17.3
20.4
12.7
10.6
5.1
13
9.1
N/A
7.6
13.3
19.8
N/A
Hb level,
g/dL
16
353
101
254
77
255
125
570
570
89
60
312
54
24
695
N/A
87
21
22
28
N/A
51
Plt
count,
109/L
PTC 1-3
Unknown
PTC 1-5
PTC 1-5
Splice
PTC 1-5
Unknown
Splice
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
Splice
PTC 1-3
Kanezaki et al15
class
Karyotype
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
47, XY, 21c
21c
47, XY, t(5;11)(p15;q13),
Unknown
Unknown
Unknown
ML-DS
Unknown
ML-DS
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
CCR
ML-DS
CCR
CCR
47, XY, 21c
CCR
47, XY, 21c
CCR
Outcome
47, XY, 21c
t(1:11)(q23;q85), 21c
47, XY, der(11)t(q23p15)
14
44
35.5
36
43
60
35
75
66
34
42
79
33
88
60
32
10
29
31
41
11.4
30
40
62
29
84
70
28
39
18
27
54
25
26
23
76
25
37
50
24
Ex2
WAVE*
Mutation
type
38
45
Blast count, %
of total
nucleated count
23
Patient
no.
Table 1. Summary of GATA1 mutations and patient (continued)
2225
43
69
10
78
34
60
75
18.5
66.5
75
85
45
38
64.5
58
34
85
11
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
WAVE*
3 36
Ins Dup
Point
Del
Del
Del
18
39
Ins Dup
Dup
7 10
13
Del Ins
Point
Dup
23
Del Ins
Del
162
19 17
21
2 13
19
No. of
nucleotides
changed, bp
Del
Point
Del
Ins
Del Ins
Dup
Point
Ins Dup
Dup
Mutation
type
4549
4604
4690
4698
4733
4740
4760
4762
4744
4638
4724
4698
4540
4747
4638
4713
4698
4723
4550
4723
4722
Position of
mutation
Loss of s
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
of stop codon
of stop codon
of sop codon
Loss of start codon
amino acid sequence
of stop codon
of stop codon
of stop codon
of stop codon
Loss of start codon
of stop codon
of stop codon
Predicted effect of
mutation on RNA
0.25
0.25
0.5
0.75
0.25
0.25
0.25
0.25
Age at
diagnosis, mo
7.4
22
27.2
24.9
69.4
32.9
78.3
46.4
149.1
62.9
16
57.5
17.3
N/A
51.2
6.2
68.6
7.7
34
61
28.6
WBC count,
109/L
12.3
16.3
17
15.6
15.4
11.9
15
20.6
16.9
12.3
18.5
10.6
14
N/A
14.8
16.9
10.9
13.2
15
17.6
16.2
Hb level,
g/dL
241
108
134
154
1178
220
1800
197
131
93
48
36
173
N/A
159
25
212
78
26
149
57
Plt
count,
109/L
1st Met
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-3
PTC 1-3
PTC 1-3
PTC 1-3
PTC 1-5
PTC 1-3
PTC 1-5
1st Met
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-3
1st Met
PTC 1-3
PTC 1-3
Kanezaki et al15
class
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
21qter(D21S1446 3)
47, XY,21.ish
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
Karyotype
Unknown
Unknown
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Outcome
ALFORD et al
31
45
Blast count, %
of total
nucleated count
2226
Patient
no.
Table 1. Summary of GATA1 mutations and patient characteristics for TMD samples (continued)
91
16.5
14
78
60.5
16
12
95
86
68
72
30
72
73
74
75
76
77
78
79
80
81
82
83
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
WAVE*
Del
Del
Point
Point
Ins Dup
Del
Dup
Point
Del
Dup
Del
Ins
Ins Dup
Point
Point
Del
Point
Ins
Mutation
type
12
67
51
19
1 12
34
No. of
nucleotides
changed, bp
4741
4733
4769
4633
4708
4706
4734
4734
4766
4722
4653
4647
4742
4768
4552
4693
4597
4654
Position of
mutation
of top codon
of stop codon
amino acid sequence
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
amino acid sequence
of stop codon
Predicted effect of
mutation on RNA
0.5
0.5
11
0.75
0.25
0.25
Age at
diagnosis, mo
52
49.8
100
107.5
410
23.2
8.5
25.4
91.2
3.39
8.9
10.6
27.6
23.83
133.9
17.3
8.4
31.6
WBC count,
109/L
13.9
14.2
11.5
13.1
11.1
14.2
10.8
13.9
13.6
10.5
12.5
10.7
14.6
9.9
13.8
13.1
9.3
16.4
Hb level,
g/dL
71
43
326
70
261
379
52
98
33
91
120
294
46
154
32.9
270
105
158
Plt
count,
109/L
PTC 1-3
PTC 1-3
Splice
Unknown
PTC 1-5
PTC 1-5
PTC 1-3
Unknown
Splice
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-3
Splice
Unknown
PTC 1-5
Unknown
PTC 1-5
Kanezaki et al15
class
N/A
N/A
N/A
47,XX,21c2
(p15;q11),12,21c17,
der(11)t(11;12)
t(10;12)(q21;p12),
(6;10)(q16;q22),der(10)
der(6)del(6)(p12p22)t
(p21)del(6)(q24,
47,XX,der(6)del(6)
N/A
N/A
N/A
N/A
(Q10;Q10) 21c
47, XY, t(21;21)
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
Karyotype
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
Outcome
32
71
5.5
87
69
70
22
3.5
32
Blast count, %
of total
nucleated count
68
67
66
Patient
no.
2227
78
79
46.5
43
15
16.5
66
38.5
58
83
20
43
49.9
61
53.5
34
25
60
51
15.5
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
WAVE*
Del Ins
Point
Dup
Ins
Del
Ins
Dup
Del
Point
Dup
Del Del
Point
Point
Point
Dup
Dup
Del
Ins
Del
Del
Ins
Substitution
Ins
Mutation
type
32
13
12
15 1
11
43
40
No. of
nucleotides
changed, bp
Unknown
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
Unknown
of stop codon
Predicted effect of
mutation on RNA
4742
4748
4707
4753
4706
4598
4720
4638
4769
4714
of stop codon
Synonymous mutation
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
4888 4907 Frameshift and introduction
92 126
4768
4742
4723
4604
4725
4638
4637
4736
4545
4654
Position of
mutation
0.25
0.75
0.5
0.25
0.25
0.5
0.25
0.5
0.25
0.25
Age at
diagnosis, mo
64.4
16.6
23.8
19.9
15.4
51
91.6
7.06
49.8
14.12
114.7
5.6
20.7
20.2
26.1
24.2
11
N/A
23.3
100
72.7
28
WBC count,
109/L
13.8
12.5
12.9
18.3
17.1
18.3
16.6
12
17
18.6
16.6
14.3
14.1
12.3
16.5
13.2
24.2
N/A
17.5
10.9
19.4
12.8
Hb level,
g/dL
547
567
265
115
62
56
150
64
209
15
141
67
901
12
48
78
158
N/A
80
70
606
274
Plt
count,
109/L
PTC 1-3
Unknown
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-5
Splice
PTC 1-5
PTC 2
Unknown
Splice
PTC 1-3
PTC 1-3
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-3
Unknown
PTC 1-5
Kanezaki et al15
class
N/A
47,XX,der(7), 212
47,XX,215/
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
47,XX,21c
N/A
N/A
47,XY,21c15
N/A
N/A
N/A
N/A
Karyotype
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
ML-DS
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
Unknown
Outcome
ALFORD et al
35
84
Blast count, %
of total
nucleated count
2228
Patient
no.
N/A
22
57.5
62
45
73
N/A
77
108
109
110
111
112
113
114
115
35
30
13.5
126
127
128
129
133
134
N/D
N/D
N/D
N/D
N/D
N/D
N/D
Ex 3.1
Ex 3.1
Ex2
Ex2
Ex2
Not found
Not found
Not found
Not found
Not found
Not found
Not found
Dup
Del
Not found
Not found
Not found
Not found
Not found
Not found
Not found
Not found
Not found
Ins
Dup
Del
Dup
Del
Del
Dup
Point
Ins
Dup
Dup
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
16
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
10
12
105
11
19
No. of
nucleotides
changed, bp
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
4778
4871
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
4733
4719
4586
4706
4672
4764
4698
4769
4731
4735
4722
Position of
mutation
N/A
N/A
N/A
N/A
N/A
N/A
N/A
of stop codon
of stop codon
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
of stop codon
of stop codon
Predicted effect of
mutation on RNA
0.5
0.75
0.25
0.5
2.5
0.38
1.5
N/A
1.5
1.5
N/A
Age at
diagnosis, mo
4.15
N/A
15.2
22.3
8.3
10.6
8.91
21.7
33.5
12.5
13.4
265.3
6.86
20.7
21.4
20.4
4.2
170
N/A
195
N/A
21.9
64.3
74.7
N/A
43.5
WBC count,
109/L
13
N/A
20.8
21
8.1
18.9
18.4
13
9.1
16.1
11.9
18
13
6.4
16.1
15
13
12.9
10.6
N/A
12
N/A
6.7
11.8
N/A
10.7
15.4
Hb level,
g/dL
100
N/A
276
60
100
65
46
24
695
193
371
353
1000
83
63
218
119
25
89
655
N/A
150
N/A
102
427
554
N/A
45
128
Plt
count,
109/L
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
PTC 2
PTC 2
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
PTC 1-3
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-5
Splice
PTC 1-5
Splice
PTC -3
PTC 1-3
PTC 1-3
Kanezaki et al15
class
Outcome
Unknown
N/A
N/A
N/A
N/A
N/A
N/A
N/A
47,XX,21c17
N/A
N/A
N/A
47, XY, 21c
Unknown
Unknown
Unknown
Unknown
Unknown
CCR
CCR
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
47, XY, 21c

N/A
Unknown
Unknown
47, XY, 21c
Unknown
Unknown
ML-DS
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Known
47, XY, 21c
N/A
47, XX, del(16), 21c
21c8/47, XY,21c7
48, XY, Y(q12),11,
N/A
N/A
N/A
N/A
47,XY,21c
N/A
N/A
N/A
N/A
N/A
Karyotype
12
132
39
125
20
124
130
80
123
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
WAVE*
Mutation
type
131
30
121
122
119
10
118
120
30
117
0.5
21
107
116
50
Blast count, %
of total
nucleated count
106
Patient
no.
2229
15
84
90
20
N/A
14
N/A
N/A
50
N/A
N/A
60
10
11
12
13
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Point
Point
Del
Point
Dup
Point
Del
Del
Point
Dup
Ins
Dup
Del
Mutation
29
20
10
13
20
11
13
56
Size, bp
4768
4768
4669
4768
4728
4548
4767
4753
4549
4722
4724
4709
4750
of stop codon
of stop codon
of stop codon
of stop codon
Loss of start codon
of stop codon
of stop codon
of stop codon
of stop codon
Predicted effect of
mutation on RNA
60
24
31
1.2
24
21
20
36
23
N/A
24
Age at
diagnosis, mo
17.4
33.4
14
2.8
11.9
4.1
27.8
40
3.16
8.5
27.7
5.3
12.6
WBC
count,
109/L
11
2.9
13
9.7
7.5
2.1
5.4
8.9
11
11.3
8.5
7.6
5.7
Hb level,
g/dL
32
3.6
112
21
18
85
27
36
20
19
22
22
Plt
count,
109/L
Splice
Splice
PTC 1-5
Splice
PTC 1-3
Splice
PTC 1-3
PTC 1-3
1st Met
PTC 1-3
PTC 1-3
PTC 1-5
PTC 1-3
Kanezaki et al15
class
Karyotype
21, 22
13, 14, 18, 19,
der(12)(t1;12)(q21;p13),
i(7)(q10), 8,
3?adk1(5)(q33),
del(1)(q21;q25), 2,
5557, XX,
XX,21c
13,21c9/47,
der(7)add(7)(p?),
46,XX,
47, XY, 21, 8
19
21c16/ 47, XX, 21c
48, XX, Add(8)(p23), 11,
21c
add(1)(q24), del(5)(p13),
47,XX,
21c, mar7
47XX,i(7)(q10),16,
47, XY, 21c
21
(q10),del(16)(q22q24),
del(3)(q21q26),I(7)
(p32)dup(1)(q21q42),
48, XY, der(1)add(1)
48, XY, 8, 21
(p13), 21, 21c7,
del(19)
del(17)(q?2224), 19,
13, 14, 14, 16,
56, XX, 2, 6, 8, 13,
47, XX, 21c
47, XX, 21c
21c
47, XY, Del(7)(q32), 8,
Died
Died
CCR
CCR
CCR
CCR
Died
Died
CCR
CCR
CCR
CCR
CCR
Outcome
ALFORD et al
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.
70
WAVE*
Position of
mutation
2230
Blast count, %
Patient
of total
no.
nucleated count
Table 2. Summary of GATA1 mutations and patient characteristics for ML-DS samples
37
16
N/A
36
93
20
23
21
44
50
16
20
16
17
18
19
20
21
22
23
24
25
26
27
N/D
N/D
N/D
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Del
Dup
Dup
Del
Del
Del
Dup
Del
Ins
Del
Ins
Point
Ins
Ins
Point
Mutation
15
10
23
35
Size, bp
4698
4715
4710
4638
4705
4638
4717
4705
4769
4684
4632
4597
4693
4736
4768
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
of stop codon
Predicted effect of
mutation on RNA
15
11
12
13
11
24
N/A
36
17
31
N/A
25
26
Age at
diagnosis, mo
6.4
3.5
7.3
8.3
6.4
32
5.4
200.3
9.8
29
1.7
N/A
42.8
11.2
WBC
count,
109/L
13.4
7.6
11.9
N/A
N/A
15.1
8.7
3.7
4.7
11.5
8.7
N/A
8.2
9.7
Hb level,
g/dL
70
60
30
N/A
N/A
5.1
22
23
24
14
N/A
33
28
Plt
count,
109/L
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
Splice
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-3
Splice
Kanezaki et al15
class
Karyotype
N/A
del(7)(q?31q?33)
(q?21;q?21),
idem,t(4;15),
47,XY,21c 2/47,
XY,21c5
21cp7/47,
?19,21c,
4749,XY,
22 6
50, XY, 10, 21c, 21,
21c, 21 5
48, XX, del (6) (q?13q?21)
21
47, XX, del (6), del (21),
21c6/ 47, XY, 21c14
(p15;q13), add(9)(q34),
47, XY, t(5;11)
47, XY, 21, 8
21c19
(q25;q25), del(6)(q13;q22),
47, XX, t(3;17)
48, XX, 8, 21
47 XX, 21c
48, XX, 8, 21c
N/A
5, 47, XY, 21c3
?del(6)(q2?5; q2?7), 21c
t(2;4)(q31;q31),
idem, der (4),
(p11.2,p21), 21c 2, 47,
47, XY, dup(X)
47. XX. 21c36
Unknown
Unknown
CCR
CCR
CCR
Died
CCR
Died
CCR
CCR
Died
Unknown
Relapsed
CCR
Unknown
Outcome
not done.
8.5
78
15
Ex2
WAVE*
Position of
mutation
28
20
14
Blast count, %
Patient
of total
no.
nucleated count
Table 2. Summary of GATA1 mutations and patient characteristics for ML-DS samples (continued)
2231
50.5
10.5
36
34
55
35
38
43
15
16
34
35
36
37
38
39
40
41
42
43
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
Dup
Dup
Dup
Del ins
Dup
Del Ins
Del
Dup
Point
Ins
Ins Dup
Del
Del
Del
Del
Mutation
21
24 4
31 6
22
2 14
Size, bp
4717
4668
4736
4683
4737
4783
4704
4734
4768
4768
4722
4604
4638
4638
4704
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
Loss of splice donor site
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
Predicted effect of
mutation on RNA
36
20
13
31
31
31
22
14
35
28
21
13
27
24
13
Age at
diagnosis, mo
160
8.1
4.9
6.1
4.2
1.9
15.1
12.4
3.4
1.2
26.4
8.5
26.4
3.9
WBC
count,
109/L
12.7
6.7
9.8
9.2
6.9
9.6
7.2
10.4
7.2
7.7
8.5
9.1
11.2
Hb level,
g/dL
1.5
219
38
16
20
22
257
144
17
109
151
107
26
147
47
Plt
count,
109/L
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-3
Splice
PTC 1-5
PTC 1-3
Splice
Splice
PTC 1-3
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
Kanezaki et al15
class
Karyotype
N/A
10}
(1;8)(q32;q22)
(q32;q22),der(8)t
2/48,idem,t(1;8)
idem,t(1;8)(q32;q22)
47,XY,21c,3/47,
47,XX,21c
N/A
N/A
N/A
N/A
48,XY,8,21c
47,XX,21c
XX,21c
mar2cp8/47,
21,21c, mar1,
(5)(p1?5),8,
4748,XX,?add
N/A
N/A
N/A
47,XY,21c25
XX,21c9
21c2/47,
(12)t(1;12)(q12;p12),
XX,8,der
(q23q25),21c4/48,
(q12;p14)?del(1)
47,XX,der(5)t(1;5)
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Outcome
ALFORD et al
not done.
19.5
33
39
31
4.5
93
30
32
31
WAVE*
Position of
mutation
2232
29
Blast count, %
Patient
of total
no.
nucleated count
16
12
10.5
17
N/A
23
51
52
53
54
55
56
57
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
WAVE*
Not Found
Point
Dup
1
16
Unknown
20
31
Del ins
Dup
21
Size, bp
Del ins
Point
Del
Point
Del
Point
Ins
Point
Ins
Mutation
4767
4734
Unknown
4727
4654
4619
4552
4702
4768
4546
4535
4736
4956
4708
Position of
mutation
of stop codon
Unknown
of stop codon
of stop codon
of stop codon
amino acid sequence
of stop codon
Loss of Start codon
Unknown
of stop codon
Synonymous mutation
of stop codon
Predicted effect of
mutation on RNA
26
N/A
21
16
15
12
20
12
19
31
5.1
N/A
5.9
4.8
2.9
3.5
3.4
2.74
6.2
22
2.5
12.9
31
6.88
WBC
count,
109/L
16
10
Age at
diagnosis, mo
13.3
N/A
11.4
12
3.3
9.2
11.1
10.7
12.1
10.6
8.5
3.1
8.1
9.7
Hb level,
g/dL
11
N/A
42
59
17
47
27
110
45
131
37
76
101
35
Plt
count,
109/L
Splice
PTC 1-3
Unknown
PTC 1-3
PTC 1-5
PTC 1-5
Unknown
PTC 1-5
Splice
1st Met
Unknown
PTC 1-3
Unknown
PTC 1-5
Kanezaki et al15
class
47,XY,21c4
21,21ccp6/
8,?14,
4850,XY,
N/A
47,XX,21c
N/A
21c4
/47,XX,
mar14
(p?q?),21c,
48,XX,der(1)
215
(q10;q10)c,
46,XY,der(21;21)
216/
(q10;q10)c,
(q10),der(21;21)
46,XY,i(7)
XY,21c8
21c2/47,
(q2425),
47,XY,?der(11)
XX,21c7
inc5/47,
14,21,21c,
50,XX,8,
N/A
47,XY,212
21,2113/
t(11;11)(p15;q13),
48,XY,i(7)(q10),
48, XY, 14, 21
N/A
N/A
N/A
Karyotype
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Outcome
not done.
15
15
49
50
14
13
47
48
11
28
45
46
24
44
Blast count, %
Patient
of total
no.
nucleated count
2233
16
23
76
67
68
69
70
42
74
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
Not Found
Point
Point
Del
Del
Del
Del
Del
Point
Dup
Unknown
16
42
Del
Unknown
Not Found
Del ins
Del
Del
Del
11
Point Del
Unknown
4713
4637
4656
4638
4638
4638
4737
4767
4719
4656
4656
Unknown
4642
4548
4698
Unknown
Synonymous mutation
Unknown
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
Unknown
of stop codon
Unknown
of stop codon
of stop codon
4637 4639 Frameshift and introduction
24
19
N/A
13
17
13
35
26
32
20
32
11
19
15
11
11
37
Age at
diagnosis, mo
5.4
4.1
N/A
3.3
23
3.2
2.8
3.3
5.46
5.6
4.8
3.7
2.6
1.8
16
6.3
6.3
WBC
count,
109/L
10.3
9.5
N/A
11.4
6.5
9.5
7.1
13.6
12
7.4
10.7
5.8
9.3
10.3
11.4
Hb level,
g/dL
21
13
N/A
46
28
25
20
69
52
19
17
130
109
43
38
84
13.7
Plt
count,
109/L
Unknown
Unknown
Unknown
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-5
PTC 1-3
Splice
PTC 1-3
PTC 1-5
PTC 1-5
Unknown
PTC 1-5
Unknown
PTC 1-5
Unknown
Kanezaki et al15
class
Karyotype
21c15
5/47,XY,
?der(22)(p12)
47,XY,21c,
/47,XY,21c12
21c,inccp2
4849,XY,21,
N/A
47,XX,21c17
7,21ccp11
(p1415;q12),
der(5)t(5;7)
(q3334;q14),
46,XY,t(4;13)
47,XY,21c10
48,XX,8,21c
N/A
N/A
N/A
47,XX,21c6
47,XX,21c8
48,XX,8,21c3/
N/A
47,XY,211
N/A
8,21c7
(p22q11),
(p11p14),r(7)
48,XY,del(5)
47,XY,21c9;
47,XX,21c5
del(7)(p12),21c7/
47,XX,?del(2)(q35),
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Outcome
not done.
N/A
20
72
73
3.5
19.5
66
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
Size, bp
Mutation
Predicted effect of
mutation on RNA
ALFORD et al
71
46
2.5
64
65
61
20.5
35
60
63
13
59
62
21
WAVE*
Position of
mutation
2234
58
Blast count, %
Patient
of total
no.
nucleated count
96
10.5
26
11
13
26
N/A
17.5
45
70.5
31
76
77
78
79
80
81
82
83
84
85
86
87
88
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
N/D
WAVE*
Del
Del
Dup
Del
Dup
Dup
Ins
Del
Point
Point
Point
Not Found
Point
Dup
Mutation
10
48
Unknown
13
Size, bp
4638
4638
4723
4638
4771
4714
4725
4638
4633
4768
4956
Unknown
4737
4744
Position of
mutation
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
of stop codon
amino acid sequence
Synonymous mutation
Unknown
amino acid sequence
of stop codon
Predicted effect of
mutation on RNA
29
16
14
11
N/A
14
11
16
43
20
17
22
26
Age at
diagnosis, mo
11.4
32.5
11.5
4.7
N/A
8.75
3.4
3.3
6.35
5.2
124
2.7
WBC
count,
109/L
13.2
7.9
8.6
9.3
N/A
11.4
10.2
10.2
10.5
12
6.4
10.6
5.9
11.8
Hb level,
g/dL
57.4
103
44
83
N/A
69
70
28
84
70
130
82
21
20
Plt
count,
109/L
PTC 1-5
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-3
PTC 1-5
PTC 1-3
PTC 1-5
Unknown
Splice
Unknown
Unknown
Unknown
PTC 1-3
Kanezaki et al15
class
Karyotype
(q10;q10)?c
der(21;21)
(q31;p13),
der(19)t(1;19)
(9)(q?13q33),
46,XX,del
11q23(MLLx2)100
13q34 2),
13q14(RB1 2),
21q22(AML1 2),
8q22(ETOx2),
46,XY22. nuc ish
N/A
N/A
N/A
N/A
N/A
47,XY,21c15
N/A
N/A
47,XY,21c8
11,21c7/
(1;8)(q21; p2223),
48,XY,der(8)t
47,XX,21c5
(p13),21c5/
47,XX,add(16)
21c,21
del(6)(q13q16),
47,X,X,
S1446 3)
21qter(D21
47,XY,21.ish
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Outcome
not done.
25
75
Blast count, %
Patient
of total
no.
nucleated count
2235
20
10
21
20
20
38
10
20
95
96
97
98
99
100
101
102
103
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
Ex2
N/D
N/D
N/D
N/D
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Dup
Ins
Del ins
Del
Mutation
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
10
26
22
Size, bp
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
4719
4733
4707
4706
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
of stop codon
of stop codon
of stop codon
of stop codon
Predicted effect of
mutation on RNA
12
10
21
38
23
15
33
11
17
15
22
11
17
Age at
diagnosis, mo
3.8
3.9
11.9
N/A
5.1
15.2
4.9
N/A
11
14.47
1.5
5.2
4.6
7.8
WBC
count,
109/L
8.6
10.9
7.3
N/A
7.2
9.9
8.6
9.9
4.1
10.4
13.7
7.1
7.5
11.1
Hb level,
g/dL
16
65
34
N/A
49
18
15
86
52
38
13
143
57
27
Plt
count,
109/L
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
Unknown
PTC 1-3
PTC 1-3
PTC 1-5
PTC 1-5
Kanezaki et al15
class
Karyotype
N/A
21c4
21c9/ 47, XX,
47, XX, t(1;22)(q23;p13),
N/A
CCR
Unknown
Unknown
CCR
47, XX, 21c
Unknown
46, XX, 21, 7
N/A
del(16)(q22q24, del(20)
(q21;p11),
der(15)t(1:15)
(p11.2p21),
(p23p25), del(8)
CCR
CCR
47 XX, ?dup(3)
Unknown
48, XY, 21, 21c
CCR
CCR
Died
Unknown
Unknown
Unknown
Unknown
Outcome
47, XY, del (7), 21
(1:11)(q23;q85), 21c
(q23p15)t
47, XY, der(11)t
der(3)t(3;3)add(3)(q29)7
21c13/47, idem,
t(3;3)(p2?3;q13.3),
47, XX, t(3;3)(q21;q26),
47, XY, 21c
21c7
47,XY,
21c8/
(q12),11,
48,X,add(Y)
N/A
46,XY3/47,XY,21c2
XY,21,21c or
21c7/46,
del(13)(q1?4),
(p2?2),8,
48,XY,add(7)
ALFORD et al
not done.
16
30
28
92
93
24
91
94
33
90
7.5
WAVE*
Position of
mutation
2236
89
Blast count, %
Patient
of total
no.
nucleated count
2237
A
TMD
//
Exon 1
//
Exon 2
ATG
Exon 3
ML-DS
Po
Inse on
B
ML-DS
TMD
1%
18%
21%
n
31%
39%
Po
Inse on
22%
21%
26%
21%
C
TMD
ML-DS
3% 3% 2%
8%
12%
3% 3%
14%
Nonsense
Silent
4%
2%
Splice
Start
Stop
76%
Unknown
70%
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.
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.
2238
ALFORD et al
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2011 118: 2222-2238

doi:10.1182/blood-2011-03-342774 originally published
online June 29, 2011
Analysis of GATA1 mutations in Down syndrome transient

myeloproliferative disorder and myeloid leukemia
Kate A. Alford, Katarina Reinhardt, Catherine Garnett, Alice Norton, Katarina Bhmer, Christine von
Neuhoff, Alexandra Kolenova, Emanuele Marchi, Jan-Henning Klusmann, Irene Roberts, Henrik
Hasle, Dirk Reinhardt and Paresh Vyas
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Analysis of GATA1 mutations in Down syndrome transient myeloproliferative

Children with Down syndrome (DS) up to

lar in TMD and ML-DS. A mutation in the

type of GATA1 mutation in our series is not prognostic of which

Results and discussion

The online version of this article contains a data supplement.

BLOOD, 25 AUGUST 2011 VOLUME 118, NUMBER 8

GATA1 MUTATIONS IN TMD AND ML-DS

end of exon 2 (PTC 1-5) or in exon 3.1 (PTC-2) resulted in low

Frameshift and introduction

Loss of splice acceptor site

introducing a stop codon

amino acid sequence

Loss of start codon

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Loss of splice acceptor site

Frameshift and introduction

Frameshift and introduction

introducing a stop codon

amino acid sequence

Loss of splice acceptor site

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Loss of splice acceptor site

Frameshift and introduction

Loss of start codon

47, XY, 21c

47, XY, 21c

47, XX, 21c

47, XX, 21c

47, XY, 21c

47, XY, 21c

47, XX, 21c

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Loss of splice donor site

Frameshift and introduction

Loss of splice acceptor site

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

introducing a stop codon

amino acid sequence

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Frameshift and introduction

Loss of splice acceptor site

introducing a stop codon