CD9 predicts ETV6-RUNX1 in childhood B-cell precursor acute lymphoblastic leukemia

Blunck, Caroline B.; Terra-Granado, Eugênia; Noronha, Elda P.; Wajnberg, Gabriel; Passetti, Fábio; Pombo-de-Oliveira, Maria S.; Emerenciano, Mariana

doi:10.1016/j.htct.2018.11.007

Caroline B. Blunck

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

Eugênia Terra-Granado

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

Elda P. Noronha

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

Gabriel Wajnberg

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

Fundação Oswaldo Cruz (Fiocruz), Rio de Janeiro, RJ, Brazil

Atlantic Cancer Research Institute, Moncton, Canada

Fábio Passetti

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

Fundação Oswaldo Cruz (Fiocruz), Rio de Janeiro, RJ, Brazil

Maria S. Pombo-de-Oliveira

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

Mariana Emerenciano ^**Corresponding author at: Research Center, Instituto Nacional de Câncer (INCA), Rua André Cavalcanti, 37, Rio de Janeiro CEP: 20231-050, RJ, Brazil. E-mail address: memerenciano@inca.gov.br (M. Emerenciano).

Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil

https://orcid.org/0000-0003-2337-8420

*Corresponding author at: Research Center, Instituto Nacional de Câncer (INCA), Rua André Cavalcanti, 37, Rio de Janeiro CEP: 20231-050, RJ, Brazil. E-mail address: memerenciano@inca.gov.br (M. Emerenciano).

Conflicts of interest

The authors declare no conflicts of interest.

Figures (3)
Tables (2)

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Figure 1
FISH using TEL/AML1 Translocation, Dual Fusion probe (spectrum orange/spectrum green) showing (A) the normal pattern (2 orange and 2 green signals) and (B) the ETV6-RUNX1 gene fusion (1 orange signal, 1 green signal and two co-localizations).

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Figure 2
CD9 expression in blast cells analyzed by FC and correlation between the CD9 percentage/MFI-r and the major molecular alterations in BCP-ALL. (A) Upper dot plots show the analysis strategy used to evaluate CD9 expression in the blast population, which was defined by the R1 (viable cells) + R2 (CD45 negative/low or intermediate) and R3 regions (CD19 positive cells). (B) R4 regions represent the percentage of CD9 positive blast cells. The left dot plot represents patients with a CD9 high-percentage expression (96%) and the right dot plot represents patients with a CD9 low-percentage expression (28%). (C) CD9 expression evaluated by ratio median fluorescence intensity (MFI-r) in blast population. In the upper histogram, non-specific IgG control histogram. The middle panel represents the low CD9 MFI-r expression in a patient ETV6-RUNX1+. The last panel represents the high CD9 MFI-r expression in a patient ETV6-RUNX1−. (D) CD9 percentage according to the common alterations (high hyperdiploidy, TCF3-PBX1, BCR-ABL1, KMT2A rearrangements) in BCP-ALL. (E) CD9 MFI-r, according to the common alterations in BCP-ALL. Horizontal lines inside the bars represent the median CD9 expression percentage or MFI-r obtained from the studied cases. *Other alterations refer to MLL-r (n = 16); TCF3-PBX1 (n = 4); BCR-ABL1 (n = 1). **No alterations were identified with the screening methods used in this study.

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Figure 3
ROC curve analysis. (A) ROC curve of CD9 percentage predicting ETV6-RUNX1. (B) ROC curve of CD9 MFI-r predicting ETV6-RUNX1.

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Table 1
Demography and laboratory characteristics of patients according to ETV6-RUNX1 status.

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Table 2
ETV6-RUNX1 prediction values for CD9 percentage and MFI-r by flow cytometry.

		ETV6-RUNX1
Variables	n (%)	Positive n (%)	Negative^a a MLL -rearrangement ( n = 14); TCF3-PBX1 ( n = 4); BCR-ABL1 ( n = 1). n (%)	p value
Age (months)
0-12	17 (9.1)	3 (6.8)	14 (9.9)	0.801
13-120	155 (83.3)	38 (86.4)	117 (82.4)
>120	14 (7.5)	3 (6.8)	11 (7.7)
Gender
Female	79 (42.4)	18 (40.9)	61 (43.0)	0.862
Male	107 (57.5)	26 (59.1)	81 (57.0)	0.862
BCP-ALL
Pro-B	13 (7.0)	1 (2.3)	12 (8.5)	0.296
Common	150 (80.6)	36 (81.8)	114 (80.3)
Pre-B	23 (12.3)	7 (15.9)	16 (11.3)
WBC (×10 ⁹ /L)
<50	147 (79.0)	37 (84.1)	110 (77.5)	0.345
≥50	39 (21.0)	7 (15.9)	32 (22.5)	0.345
TOTAL	186	44 (23.7)	142 (76.3)

CD9 expression
	Percentage	MFI
All patients (n = 186)
Best cutoff^a a Best cutoff for maximizing CD9 expression sensitivity and specificity to predict ETV6-RUNX1 ; FISH: fluorescence in situ hybridization; MFI-r: median fluorescence intensity ratio.	64%	12.52
Sensitivity	99.0%	72.0%
Specificity	88.2%	86.3%
Positive predictive value	71.4%	81.8%
Negative predictive value	76.3%	89.0%
Accuracy	0.84	0.80
Time to diagnosis
<24 h (n = 121)
Best cutoff^a a Best cutoff for maximizing CD9 expression sensitivity and specificity to predict ETV6-RUNX1 ; FISH: fluorescence in situ hybridization; MFI-r: median fluorescence intensity ratio.	64%	10.53
Sensitivity	96.0%	70.0%
Specificity	87.0%	81.0%
Positive predictive value	84.0%	79.0%
Negative predictive value	87.0%	62.0%
Accuracy	0.88	0.79
Time to diagnosis
>24 h (n = 65)
Best cutoff^a a Best cutoff for maximizing CD9 expression sensitivity and specificity to predict ETV6-RUNX1 ; FISH: fluorescence in situ hybridization; MFI-r: median fluorescence intensity ratio.	75%	13.73
Sensitivity	28.0%	50.0%
Specificity	90.0%	91.0%
Positive predictive value	91.0%	87.0%
Negative predictive value	25.0%	47.3%
Accuracy	0.87	0.80

[1] *Corresponding author at: Research Center, Instituto Nacional de Câncer (INCA), Rua André Cavalcanti, 37, Rio de Janeiro CEP: 20231-050, RJ, Brazil. E-mail address: memerenciano@inca.gov.br (M. Emerenciano).

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CD9 predicts ETV6-RUNX1 in childhood B-cell precursor acute lymphoblastic leukemia

ABSTRACT

Introduction:

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