CEBPA gene mutations in Egyptian acute myeloid leukemia patients: impact on prognosis

Figures & data

Table 1. Primers used to amplify CEBPA from genomic DNA

Primer	Sequence
PP1F	5′TCGCCATGCCGGGAGAACTATAAC-3′(sense)
PP1R	5′-CTGGTAAGGGAAGAGGCCCGCCAG-3′(antisense)
PP2F	5′-CCGCTGGTGATCAAGCAGGA-3′ (sense)
PP2R	5′-CACG GTCTGGGCAAGCCTCGAGAT-3′ (antisense)

Figure 1. SSCP of CEBPA gene (P1): C, normal control. Lanes 1–8: samples from eight different AML patients. Lane 8 represent homozygous mutant CEBPA (mobility shift). Lanes 1–7 represent WT CEBPA.

Figure 2. SSCP of CEBPA gene (p2): C, normal control; 1–4, samples from four different AML patients. Lane 1 represents heterozygous mutant CEBPA gene. Lanes 2–4 represent WT CEBPA gene.

Table 2. Hematological characteristics of AML patients with CEBPA mutation

Variables	Total (n = 55)	Wild-type CEBPA (n = 44)	Mutant CEBPA (n = 11)	P
Age (years)	45.65 ± 16.276	44.55 ± 14.92	46.09 ± 21.76	0.939
Sex				0.176
Male	25.0 (45.5%)	22.0 (50%)	3.0 (27.3%)
Female	30.0 (54.5%)	22.0 (50%)	8.0 (72.7%)
TLC (×10^9/l)	79.26 ± 93.5	77.24 ± 92.64	87.35 ± 101.29	0.752
Hemoglobin level (g/dl)	7.7 ± 1.5967	7.48 ± 1.51	8.59 ± 1.69	0.037*
Platelet count (×10^9/l)	57.920 ± 69.99	61.47 ±77.39	43.7 ± 20.48	0.457
Peripheral blasts (%)	62.3 ± 24.7	60.55 ± 24.33	69.55 ± 26.01	0.284
Marrow blasts (%)	69.65 ± 20.95	70.05 ± 20.77	68.09 ± 22.61	0.785
LDH (U/l)	929.35 ± 746.7	967.70 ± 786.874	226.4 ± 562.1	0.003*
FAB types				0.719
M0	2.0 (3.6%)	2.0 (4.5%)	0 (0%)
M1	4.0 (6.2%)	3.0 (6.8%)	1.0 (9.1%)
M2	11.0 (16.9%)	8.0 (18.2%)	3.0 (27.2%)
M3	4.0 (6.2%)	4.0 (9.1%)	0 (0%)
M4	18.0 (27.7%)	13.0 (29.5%)	5.0 (45.5%)
M5	12.0 (18.5%)	11.0 (25.0%)	1.0 (9.1%)
M6	4.0 (6.2%)	3.0 (6.8%)	1.0 (9.1%)
Immunophenotyping
cMPO	43.0 (78.2%)	33.0 (75%)	10.0 (90.9%)	0.253
CD13	47.0 (85.5%)	38.0 (86.4%)	9.0 (81.8%)	0.702
CD33	51.0 (92.7%)	40.0 (90.9%)	11.0 (100%)	0.299
CD117	44.0 (80%)	35.0 (79.5%)	9.0 (81.8%)	0.866
HLA-DR	30.0 (54.5%)	25.0 (56.8%)	5.0 (45.5%)	0.498
CD14	29.0 (52.7%)	23.0 (52.3%)	6.0 (54.5%)	0.893
CD11b	26.0 (47.3%)	21.0 (47.7%)	5.0 (45.5%)	0.893

Mean ± SD are indicated in TLC, hemoglobin, platelets, peripheral blasts, marrow blasts, and LDH. Number and percentage of cases are shown in FAB, number and percentage of positive cases are shown in immunophenotyping.

*P value <0.05 is considered as significant.

Table 3. Cytogenetics of AML patients with CEBPA mutation

Variables	Total (n = 55)	Wild-type CEBPA (n = 44)	Mutant CEBPA (n = 11)	P value
Cytogenetics				0.007*
Normal karyotype	30.0 (54.5%)	20.0 (45.5%)	10.0 (90.9%)
Abnormal cytogenetics	25.0 (45.5%)	24 (54.5%)	1.0 (9.1%)
Cytogenetic risk				0.179
Favorable	18.0 (32.7%)	17.0 (38.6%)	1.0 (9.1%)
Intermediate	32.0 (58.2%)	22.0 (50.0%)	10.0 (90.9%)
Unfavorable	5.0 (9.1%)	5.0 (11.4%)	0 (0%)

Number and percentage of cases are shown in cytogenetics and cytogenetic risk.

*P value <0.05 is considered as significant.

Table 4. Clinical outcome in relation to CEBPA gene mutation status

	Wild-type CEBPA (n = 44)	Mutant CEBPA (n = 11)	P value
Complete remission	26.0 (59.1%)	10.0 (90.9%)	0.047*
Refractory	9.0 (20.5%)	1.0 (9.1%)	0.382
Relapse	4.0 (9.1%)	0 (0%)	0.299
Death	18.0 (40.9%)	1.0 (9.1%)	0.047*

*P value <0.05 is considered as significant.

Table 5. Survival in CEBPA mutations versus non-CEBPA mutation in AML patients

	Wild-type CEBPA (n = 44)		Mutant CEBPA (n = 11)		Significance Log rank (Mantel-Cox)
	Cumulative survival	Median (CI 95%)	Cumulative survival	Median (CI 95%)
Disease free survival	27.3%	21.0 (16.67–25.33)	54.5%	26.0 (15.21–36.79)	0.005*
Overall survival	52.3%	25.0 (23.42–26.58)	63.6%	27.0 (16.21–37.79)	0.013*

CI 95%, confidence interval at 95%; cumulative survival, cumulative proportion surviving at 24 months.

*P value <0.05 is considered as significant.

Figure 3. Kaplan–Meier estimates of DFS. DFS among CEBPA^double-mut versus CEBPA WT AML (P = 0.009) and versus CEBPA^single-mut AML (P = 0.163); CEBPA^single-mut AML versus CEBPA^wt AML (P = 0.634; pooled = 0.024).

Figure 4. Kaplan–Meier estimates of OS. OS among CEBPA^double-mut versus CEBPA^wt AML (P = 0.008) and versus CEBPA^single-mut AML (P = 0.197); CEBPA^single-mut AML versus CEBPA^wt AML (P = 0.712; pooled = 0.021).