Gene expression of hENT1, dCK, CDA, dCMPD and topoisomerase IIα as an indicator of chemotherapy response in AML treated with cytarabine and daunorubicin

Figures & data

Table 1 Baseline characteristics of the study population (n=90)

Parameters (n=90)	N	Percent
Age groups, years  <15  15–40  41–60  >60	3 62 24 1	3.3 68.9 26.7 1.1
Gender  Male  Female	66 24	73.3 26.7
AML classification (WHO)  APML (M3)  AML without maturation (M1)  AML with maturation (M2)  Others Translocation 6:9  AML with minimal differentiation (M0)  Acute myelomonocytic leukemia (M4)  Acute panmyelosis with fibrosis  Myeloid proliferations related to Down syndrome  Unknown	17 15 44 8 2 2 2 1 1 6	18.9 16.7 48.9 8.9 6.7
MPO status  Negative  Positive  Unknown	14 62 14	15.6 68.9 15.6
FLT3 mutation  Negative  Positive  Unknown	35 7 48	38.9 7.8 53.3
NPM1 mutation  Negative  Unknown	13 77	14.4 85.6)
Karyotyping  Unfavorable  Favorable t(15:17)  Normal  Unknown	18 7 24 41	20.0 7.8 26.7 45.6
Therapeutic response  Resistant  Relapse  Persistant remission	34 19 37	37.8 21.1 41.1
Final outcome  Poor  (resistant + relapse)  Good (persistent remission)	53 37	58.9 41.1
Survival status  Died  Alive  Unknown	42 44 4	46.7 48.9 4.4

Abbreviations: AML, acute myeloid leukemia; MPO, myeloperoxidase.

Table 2 Comparison between groups according to baseline characteristics

Parameters	Groups	N	χ^2 value	P-value	OR	95% CI
						Lower	Upper
AML classification (APML vs others)
Gender	Male	61	0.16	0.69	1.29	0.37	4.45
	Female	23
MPO	Negative	12	13.69	<0.001	11.20	2.61	47.99
	Positive	61
FLT3	Negative	32	0.01	1.00	1.11	0.11	11.33
	Positive	7
MLL	Negative	9			Invalid
	Positive	5
Remission status	Relapse	16	6.32	0.01	0.10	0.01	0.81
	Remission	34
Survival status	Dead	39	4.28	0.05	0.29	0.08	0.98
	Alive	42
Final response	Poor	50	15.51	<0.001	0.09	0.02	0.35
	Good	34
MPO status (negative vs positive)
Gender	Male	54	0.38	0.54	0.68	0.20	2.32
	Female	22
Classification	APML (M3)	11	13.69	<0.001	11.20	2.61	47.99
	Others	62
FLT3	Negative	30	1.44	0.56	Invalid
	Positive	6
MLL	Negative	9	0.60	1.00	Invalid
	Positive	5
Remission status	Relapse	18	5.18	0.03	0.11	0.01	0.97
	Remission	29
Survival status	Dead	38	1.05	0.31	0.53	0.16	1.81
	Alive	36
Final response	Poor	47	8.05	0.01	0.18	0.05	0.64
	Good	29
Remission (relapse vs persistent remission)
Gender	Male	45	0.04	1.00	0.88	0.22	3.46
	Female	11
Classification	APML (M3)	15	6.32	0.02	0.10	0.01	0.81
	Others	35
MPO status	Negative	11	5.18	0.03	0.11	0.01	0.97
	Positive	36
FLT3	Negative	26			Invalid
	Positive	–
MLL	Negative	7	0.74	1.00	Invalid
	Positive	2
Survival status	Dead	19	23.92	<0.001	28.13	6.16	128.36
	Alive	34
Final response	Poor	19	56.00	<0.001	Invalida
	Good	37
Final response (poor vs good)
Gender	Male	66	1.93	0.16	0.49	0.18	1.35
	Female	24
Classification	APML (M3)	17	15.51	<0.001	0.09	0.02	0.35
	Others	67
MPO status	Negative	14	8.05	0.01	0.18	0.05	0.64
	Positive	62
FLT3	Negative	35	5.17	0.03	Invalid
	Positive	7
MLL	Negative	10	0.13	1.00	0.67	0.08	5.88
	Positive	5
Remission status	Relapse	19	56.00	<0.001	Invalida
	Remission	37
Survival status	Dead	42	30.93	<0.001	20.36	6.07	68.26
	Alive	44
Survival status (dead vs alive)
Gender	Male	63	1.82	0.18	0.51	0.19	1.36
	Female	23
Classification	APML (M3)	16	4.28	0.05	0.29	0.08	0.98
	Others	65
MPO status	Negative	13	1.05	0.31	0.53	0.16	1.81
	Positive	61
FLT3	Negative	33	1.56	0.41	0.33	0.06	1.97
	Positive	7
MLL	Negative	10	0.50	0.58	0.43	0.04	4.64
	Positive	4
Remission status	Relapse	19	23.92	<0.001	28.13	6.16	128.36
	Remission	34
Final Response	Poor	52	30.93	<0.001	20.36	6.07	68.26
	Good	34

Note:

a Because all relapse in poor and all remission in good category. All chi-squared values df =1

Table 3 Comparison between groups according to gene expression

Parameters	Groups	N	χ^2 value	P-value	OR	95% CI
						Lower	Upper
Remission status (relapse vs persistent remission)
Bone marrow:
dCK	Low (<1)	42	0.26	0.68	0.67	0.14	3.24
	High (>1)	8
CDA	Low (<1)	27	0.00	0.95	0.96	0.29	3.24
	High (>1)	23
dCMPD	Low (<1)	40	2.38	0.12	0.33	0.08	1.39
	High (>1)	10
hENT1	Low (<1)	23	1.38	0.24	0.47	0.13	1.67
	High (>1)	27
dCK/CDA	Low (<1)	37	0.40	0.73	1.60	0.37	6.91
	High (>1)	13
Topo-IIα	Low (<1)	4	0.83	0.57	2.54	0.32	19.96
	High (>1)	46
Peripheral
blood:
dCK	Low (<1)	35	1.37	0.30	2.66	0.50	14.25
	High (>1)	11
CDA	Low (<1)	24	0.01	0.91	1.07	0.31	3.68
	High (>1)	22
dCMPD	Low (<1)	32	1.15	0.33	2.20	0.51	9.51
	High (>1)	14
hENT1	Low (<1)	19	3.21	0.07	3.15	0.88	11.31
	High (>1)	27
dCK/CDA	Low (<1)	33	1.51	0.22	0.44	0.12	1.66
	High (>1)	13
Topo-IIα	Low (<1) High (>1)	8 38	13.28	0.001	26.25	2.81	245.52
Final response (poor vs good)
Bone marrow:
dCK	Low (<1)	66	1.06	0.30	0.55	0.17	1.74
	High (>1)	16
CDA	Low (<1)	43	0.08	0.77	0.88	0.37	2.11
	High (>1)	39
dCMPD	Low (<1)	61	4.11	0.04	0.32	0.11	0.99
	High (>1)	21
hENT1	Low (<1)	33	3.18	0.07	0.44	0.18	1.09
	High (>1)	49
dCK/CDA	Low (<1)	63	1.00	0.32	1.69	0.60	4.74
	High (>1)	19
Topo-IIα	Low (< 1)	5	0.02	1.00	1.13	0.18	7.12
	High (>1)	77
Peripheral
blood:
dCK	Low (<1)	61	2.15	0.14	2.28	0.75	6.97
	High (>1)	16
CDA	Low (<1)	43	0.38	0.54	1.33	0.53	3.33
	High (>1)	34
dCMPD	Low (<1)	49	0.02	0.90	0.94	0.36	2.42
	High (>1)	28
hENT1	Low (<1)	35	3.65	0.06	2.50	0.97	6.47
	High (>1)	42
dCK/CDA	Low (<1)	57	0.31	0.58	0.74	0.26	2.13
	High (>1)	20
Topo-IIα	Low (<1)	14	7.80	0.01	11.82	1.46	95.85
	High (>1)	63
Survival status (dead vs alive)
Bone marrow:
dCK	Low (<1)	64	0.31	0.58	0.73	0.23	2.24
	High (>1)	15
CDA	Low (<1)	42	0.02	0.88	1.07	0.44	2.59
	High (>1)	37
dCMPD	Low (<1)	58	0.35	0.55	0.74	0.27	2.01
	High (>1)	21
hENT1	Low (<1)	31	0.05	0.82	1.11	0.45	2.74
	High (>1)	48
dCK/CDA	Low (<1)	60	0.23	0.64	1.29	0.45	3.65
	High (>1)	19
Topo-IIα	Low (<1)	5	0.10	1.00	0.74	0.12	4.71
	High (>1)	74
Peripheral
blood:
dCK	Low (<1)	59	1.47	0.23	2.07	0.63	6.79
	High (>1)	15
CDA	Low (<1)	41	0.08	0.78	1.14	0.46	2.86
	High (>1)	33
dCMPD	Low (<1)	47	0.14	0.71	1.20	0.46	3.10
	High (>1)	27
hENT1	Low (<1)	33	2.52	0.11	2.12	0.83	5.39
	High (>1)	41
dCK/CDA	Low (<1)	55	0.29	0.59	0.75	0.26	2.13
	High (>1)	19
Topo-IIα	Low (<1)	14	0.14	0.71	0.80	0.25	2.59
	High (>1)	60

Notes: All chi-squared values df =1.

Abbreviations: CDA, cytidine deaminase; dCK, deoxycytidine kinase; dCMPD, deoxycytidine monophosphate deaminase; hENT1, human equilibrative nucleoside transporter 1; Topo-IIα, topoisomerase IIα.

Table 4 Spearman correlation matrix

Variables		dCK_M	dCK_B	CDA_M	CDA_B	dCMPD_M	dCMPD_B	hENT1_M	hENT1_B	Topo-IIα _M	Topo-IIα _B	dCK/CDA_M	dCK/CDA_B
dCK_M	rs	1.00	0.52a	0.15	0.04	0.37a	0.13	0.41a	0.30b	0.13	0.04	0.65a	0.30b
	n	82	69	82	69	82	69	82	69	82	69	82	69
dCK_B	rs		1.00	0.00	0.09	0.09	0.35a	0.15	0.47a	–0.11	0.15	0.43a	0.73a
	n		77	69	77	69	77	69	77	69	77	69	77
CDA_M	rs			1.00	0.54a	0.29	0.01	0.48a	0.00	0.41a	–0.24	–0.55a	–0.29
	n			82	69	82	69	82	69	82	69	82	69
CDA_B	rs				1.00	0.13	0.36a	0.18	0.41a	0.09	0.23b	–0.28b	–0.48a
	n				77	69	77	69	77	69	77	69	77
dCMPD_M	rs					1.00	0.38a	0.59a	0.19	0.23b	0.10	0.03	–0.09
	n					82	69	82	69	82	69	82	69
dCMPD_B	rs						1.00	0.17	0.68a	0.00	0.30b	0.14	0.02
	n						77	69	77	69	77	69	77
hENT1_M	rs							1.00	0.29b	0.30b	0.00	–0.08	–0.05
	n							82	69	82	69	82	69
hENT1_B	rs								1.00	–0.06	0.46a	0.37a	0.09
	n								77	69	77	69	77
Topo-IIα_M	rs									1.00	0.28b	–0.18	–0.14
	n									82	69	82	69
Topo-IIα_B	rs										1.00	0.27b	–0.06
	n										77	69	77
dCK/CDA_M	rs											1.00	0.45a
	n											82	69
dCK/CDA_B	rs												1.00
	n												77

Notes:

a P-value <0.01.

b P-value <0.05 but more than 0.01.

Abbreviations: B, peripheral blood; CDA, cytidine deaminase; dCK, deoxycytidine kinase; dCMPD, deoxycytidine monophosphate deaminase; hENT1, human equilibrative nucleoside transporter 1; Topo-IIα, topoisomerase IIα. M, bone marrow; rs, Spearman correlation coefficient; n, sample size.

Figure 1 Kaplan–Meier curves showing OS in relationship to dCK, CDA, dCK/CDA, dCMPD, hENT1 and Topo-IIα expression in bone marrow (M) and peripheral blood (B).

Abbreviations: CDA, cytidine deaminase; Cum, cumulative; dCK, deoxycytidine kinase; dCMPD, deoxycytidine monophosphate deaminase; hENT1, human equilibrative nucleoside transporter 1; OS, overall survival; Topo-IIα, topoisomerase IIα.

Figure 2 Kaplan–Meier curves showing DFS in relationship to dCK, CDA, dCK/CDA, dCMPD, hENT1 and Topo-IIα expression in bone marrow (M) and peripheral blood (B).

Abbreviations: CDA, cytidine deaminase; Cum, cumulative; dCK, deoxycytidine kinase; dCMPD, deoxycytidine monophosphate deaminase; DFS, disease free survival; hENT1, human equilibrative nucleoside transporter 1; Topo-IIα, topoisomerase IIα.

Table S1 Gene expression profile, grouped by various factors

Parameters	N	dCK			CDA			dCMPD			hENT1			Topo-IIa			dCK/CDA
		Median	25	75	Median	25	75	Median	25	75	Median	25	75	Median	25	75	Median	25	75
Bone marrow
AML classification  APML (M3) with t(15:17)  AML without maturation (M1)  AML with maturation (M2)  Others	17 14 40 11	0.03 0.05 0.03 0.01	0.00 0.00 0.00 0.00	0.90 3.10 0.61 0.29	1.29 0.52 0.50 0.44	0.02 0.01 0.06 0.00	5.52 10.85 2.71 3.17	0.77 0.97 0.59 0.13	0.39 0.29 0.10 0.06	1.49 13.47 0.90 0.44	1.71 3.26 1.50 0.47	0.04 0.00 0.02 0.18	3.02 14.44 3.64 1.08	19.27 18.25 19.53 14.82	6.61 8.80 8.86 2.54	123.5 119.4 39.95 17.49	0.35 0.42 0.11 0.19	0.00 0.00 0.00 0.00	1.33 13.44 0.96 0.96
APML vs others  APML (M3)  All others (Poor)	17 61	0.03 0.03	0.00 0.00	0.90 0.67	1.29 0.54	0.02 0.06	5.52 3.28	0.77 0.54	0.39 0.13	1.49 1.15	1.71 1.56	0.04 0.10	3.02 4.10	19.27 17.24	6.61 8.68	123.5 43.57	0.35 0.12	0.00 0.00	1.33 0.98
Myeloperoxidase status  Negative  Positive	13 58	0.03 0.04	0.00 0.00	0.68 1.00	1.27 0.52	0.04 0.01	5.31 2.90	0.38 0.65	0.08 0.26	0.79 1.33	0.47 2.14	0.04 0.33	2.76 4.30	15.72 17.33	7.74 7.03	483.5 40.34	0.15 0.26	0.01 0.00	0.66 2.10
Pre or postchemotherapy sample  Prechemo  Postchemo	32 50	0.01 0.04	0.00 0.00	0.90 0.67	0.36 1.30	0.01 0.05	1.65 6.23	0.72 0.51	0.13 0.13	1.43 0.98	1.64 1.31	0.04 0.04	3.89 3.61	13.36 18.23	6.84 7.34	87.02 61.95	0.96 0.09	0.01 0.00	3.29 0.96
Remission status  Resistant  Relapse  Remission	32 15 35	0.04 0.08 0.01	0.00 0.00 0.00	1.07 0.82 0.53	0.88 0.38 0.45	0.12 0.01 0.01	4.85 6.76 2.54	0.68 0.50 0.62	0.24 0.07 0.09	1.44 1.13 0.86	3.11 1.84 0.92	0.32 0.21 0.01	4.64 8.76 2.43	17.17 15.93 19.27	6.84 7.57 6.64	93.13 32.37 100.8	0.18 0.05 0.65	0.00 0.00 0.00	0.96 0.96 2.02
Survival  Dead  Alive	37 42	0.04 0.02	0.00 0.00	0.90 0.61	0.54 0.76	0.02 0.01	3.51 2.89	0.49 0.65	0.08 0.29	1.24 0.94	1.64 1.36	0.12 0.04	4.02 3.75	15.93 19.12	6.93 8.08	48.48 61.95	0.14 0.27	0.00 0.00	0.98 1.77
Blood
AML classification  APML (M3) with t(15:17)  AML without maturation (M1)  AML with maturation (M2)  Others	14 13 38 12	0.56 0.21 0.00 0.11	0.00 0.00 0.00 0.01	1.33 1.61 0.43 1.04	1.22 1.03 0.24 0.35	0.04 0.09 0.01 0.04	2.20 2.90 2.25 2.35	0.60 1.68 0.44 0.58	0.28 0.54 0.02 0.19	1.31 2.98 1.04 2.21	2.22 2.37 0.74 1.22	0.31 0.05 0.01 0.66	3.63 5.66 2.14 4.11	8.55 7.57 6.18 6.55	2.28 2.89 0.37 3.72	18.72 16.03 24.57 416.2	0.65 0.05 0.21 0.61	0.00 0.00 0.00 0.02	0.95 5.21 1.01 241.9
APML vs others  APML (M3)  All others (Poor)	14 58	0.56 0.00	0.00 0.00	1.33 0.66	1.22 0.52	0.04 0.03	2.20 2.45	0.60 0.53	0.28 0.15	1.31 1.52	2.22 0.98	0.31 0.02	3.63 3.23	8.55 5.56	2.28 1.68	18.72 14.55	0.65 0.06	0.00 0.00	0.95 1.46
Myeloperoxidase status  Negative  Positive	12 55	0.11 0.00	0.00 0.00	1.46 0.81	1.28 0.69	0.05 0.03	2.45 2.19	0.21 0.75	0.00 0.29	0.60 1.47	0.47 1.24	0.01 0.07	2.98 3.28	6.78 7.57	2.91 1.99	81.44 15.13	0.71 0.06	0.02 0.00	0.96 1.53
Pre or postchemotherapy sample  Prechemo  Postchemo	31 46	0.01 0.03	0.00 0.00	1.19 0.66	0.14 1.20	0.01 0.05	1.20 3.94	0.77 0.50	0.23 0.17	1.84 1.20	1.40 0.79	0.03 0.03	3.35 2.95	7.49 7.22	1.83 1.92	14.36 24.57	0.92	0.00	1.43
Remission status  Resistant  Relapse  Remission	31 15 31	0.00 0.05 0.09	0.00 0.00 0.00	0.34 0.63 1.11	0.64 0.21 0.85	0.03 0.01 0.03	1.56 2.42 2.55	0.91 0.41 0.74	0.23 0.17 0.17	1.84 0.62 1.40	0.90 0.74 1.61	0.00 0.02 0.41	3.21 4.61 2.86	4.68 1.56 9.37	1.23 0.24 3.90	14.36 57.38 23.06	0.02 0.96 0.75	0.00 0.00 0.02	0.96 4.50 0.96
Survival  Dead  Alive	35 39	0.03 0.01	0.00 0.00	0.39 1.10	0.69 0.64	0.05 0.01	2.42 2.19	0.54 0.74	0.22 0.24	1.31 1.44	0.80 1.54	0.01 0.07	3.21 3.28	5.88 7.57	2.04 1.83	23.37 15.13	0.06 0.75	0.00 0.00	1.88 0.96

Note: The values are given as median and IQR (25th–75th percentile).

Abbreviations: AML, acute myeloid leukemia; CDA, cytidine deaminase; dCK, deoxycytidine kinase; dCMPD, deoxycytidine monophosphate deaminase; hENT1, human equilibrative nucleoside transporter 1; Topo-IIa, topoisomerase IIα.