Identification of chromosomal abnormalities and genomic features in near-triploidy/tetraploidy-acute leukemia by fluorescence in situ hybridization

Figures & data

Figure 1 BM morphologic features.

Notes: (A-1) (Case 7): Blasts are large with multiple and irregular nuclei and large nucleoli; cytoplasm is dark blue and filled with granules and prominent cytoplasmic vacuoles (arrow). Peripheral blood (A-2) (Case 7): Auer rod (arrow). (B) (Case 9): Blasts are different in size with pleomorphic nuclei and prominent nucleoli. Multinucleate blasts (arrow). (C) (Case 10): Blasts are different in size and filled with granules and prominent cytoplasmic pseudopodia. Faggot cell (arrow). (D) (Case 3): Atypical large blasts with irregular nuclear outlines and frequent cytoplasmic vacuoles, showing large nuclei and nucleoli. (E) (Case 2): Blasts are large with prominent, multiple, and pleomorphic nuclei. Magnification ×1000.

Abbreviation: BM, bone marrow.

Figure 2 Karyotype and FISH analysis (Case 7).

Notes: Numbers 1–22 represent chromosomes. (A) Karyotype of diploidy (R-banding): 46, XY, t (8;21)(q22;q22). 8q-(red arrow) and 21q+ (green arrow). (B) Karyotype of tetraploidy (R-banding): 92, XXYY, t(8;21)(q22;q22)×2. 8q-(red arrow) and 21q+ (green arrow). (C) FISH analysis with GLP RUNX1–RUNX1T1 dual color fusion probe (located at 21q22/8q22). Revealing 2F4O4G signals. Two fusions on the end of 21q+, four red signals (red arrow) are on native chromosome 8 and 8q−, two green signals proximal to the centromere of 21q+ (yellow arrow), and two native chromosome 21 (green arrow). (D) FISH analysis with GLP C-MYC dual color break-apart probe (located at 8q24); the picture displays that 8q24 (MYC) (red arrow) had not moved to 21q+. (E) FISH analysis with GLP ETV6–RUNX1 dual color fusion probe (located at 12p13/21q22). Tetraploidy metaphase shows red signals (red arrow) on chromosome 12, and six green signals (green arrow) consist of two on native chromosome 21 and four on 21q+×2. Magnification ×1000.

Abbreviation: FISH, fluorescence in situ hybridization.

Figure 3 Karyotype and FISH analysis (Case 9).

Notes: Numbers 1 -22 represent chromosomes. (A) Karyotype (R-banding): 86, XXYY, −5, –5, –7, I (11q)×2, –13, −13, –18. Red arrow represents isodicentric 11q chromosomes. (B) FISH analysis with GLP MLL dual color break-apart probe (located at 11q23). The probe confirms two idic (11q) chromosomes (red arrow). Magnification ×1000.

Abbreviation: FISH, fluorescence in situ hybridization.

Figure 4 Genes change in near-triploid/tetraploid-AL in 10 cases.

Abbreviation: AL, acute leukemia.

Table 1 Demographic and clinical features and treatment outcomes in near-triploidy/tetraploidy-AL patients

Case	Age (years)/ sex	WBC (×10^9/L)	Hb (g/L)	Platelets (×10^9/L)		Juvenile cells ratio	Percent blasts	Diagnosis	Immunophenotypic analysis	Treatment	OS (months)
1	8/M	40.2	78	30	0.6		78.0	ALL (high	CD10 (+), CD19 (+),	VDLP, CAT,	28
								risk)	CD13 (+), CD33 (+),	HD-MTX, VDLD
									CD34 (+), HLADR (+)
2	45/M	1.99	110	79	0.06		78.5	ALL-B	CD34 (+), CD13 (+),	CALLG-2008	6
									CD19 (+), CD34 (+),
									HLADR (+), DT (+),
									CD79α (−), MPO (−),
									CD5 (−), CD10 (−),
									CD14 (−), CD7 (−)
3	49/F	2.0	83	84	N		72.5	ALL	CD13 (+), CD15 (+),	VLP + IA, IA, NA	5
									CD33 (+), CD15 (+)
									CD56 (−), MPO (−),
									CD34 (−), HLADR (−)
4	88/F	1.8	80	28	N		45.5	ALL	CD15 (+), CD19(−),	VP-CAM	3
									CD16 (+) CD56 (+),
									CD56 (+), cyCD3 (−)
5	42/M	18.6	36	10	0.18		40.0	AML-M2a	CD13 (+), CD15 (+),	IA, FlAG	12
									CD33 (+), CD117 (+),
									MPO (+), HLADR (+),
									CD34 (+)
6	59/F	8.1	110	7	0.51		95.5	AML-M4	CD34 (+), CD117 (+)	IA, FlAG, HAE,	10
										NAE, decitabine
										+ CAG
7	23/M	2.1	49	39	0.22		59.5	AML-M2	HLADR (+), CD33 (+),	IA, FlAG, HAA,	10 (alive)
									CD38 (+), CD56 (+),	cladribine in the
									CD117 (+), MPO (+),	regimen
									CD4 (+), CD13 (+),
									CD15 (+), CD19 (+),
									CD64 (+), CD34 (+), CR
									(−), CK5/6 (−)
8	28/M	29.96	113	30	0.92		91.2	AML-M5a	CD13 (+), CD15 (+),	IA, MD-Ara-C	12
									CD33 (+), CD117 (+),
									MPO (+), HLADR (+),
									CD34 (+)
9	35/M	4.06	82	112	0.12		50.5	AML	CD13 (+), CD33 (+),	IA	2
									CD117 (+), MPO (+),
									HLADR (+), CD34 (+)
10	6/M	2.4	70	12	N		84.0	APL(M3a)	CD13 (+), CD33 (+),	ATRA + IDA,	34 (alive)
									CD117 (+), MPO (+)	ATRA + VP-16 +
										Ara-C, ATRA +
										HA, ATRA + VP-
										16 + HD-Ara-C

Abbreviations: AL, acute leukemia; ALL, acute lymphoblastic leukemia; AML, acute myeloid leukemia; ATRA, all-trans retinoic acid; CAG, Cytarabine, Aclarubicin, and G-CSF; CALLG-2008, Guideline in China for acute leukemia; CAT, Cyclophosphamide, Ara-C, and Topotecan; F, female; FlAG, Fludarabine, Cytarabine, and G-CSF; HA, Homoharringtonine and Ara-C; HAA, Homoharringtonine, Ara-C, and Aclarubicin; HAE, Homoharringtonine, Ara-C, and Etoposide; Hb, hemoglobin; HD, high dose; IA, Cytarbine and Darubicin; IDA, Idarubicin; M, male; MD, medium dose; MTX, Methotrexate; N, not found; NA, Mitoxantrone (Novantrone) and Ara-C; NAE, Mitoxantrone (Novantrone), Ara-C, and Etoposide; VDLD, Vincristine, Dexamethasone, Levo-asparaginase, and Daunorubicin; VDLP, Vincristine, Daunorubicin, l-asparaginase, and Prednisone; VLP, Vincristine, l-asparaginase, and Prednisone; VP-16, Etoposide; VP-CAM, Vincristine, Prednisone, Cyclophosphamide, Ara-C, 6-MP (6-mercaptopurine); WBC, white blood cell.

Table 2 Karyotypes

Case	Karyotype
1	86−97<4n>, XXYY, +X, +1, +5, +8, +10, +12, +16, +20, +21, +22, −Y, −3, −4, −5, −6, −9, −11, −14, −18 [CP10]/46, XY, N [12]
2	71−74, XXY, +1, +2, −3, +4, +5, +6, del(6)(q22), + del(6)(q22), −7, +8, +12, −13, +14, +der(15), −17, −19, −20, +21, add(22)(p13), +add(22) (p13) [CP10]
3	1) 90−92, XXXX, der(1), −1, −8, −21 [cp10]a 2) 90−92, XXXX, −1, der(1), del(8)(q24), −9 [cp10]b
4	70−76, XX, −X, +1, +2, i(3q), add(9)(p13), +10, add(11)(q23), −12, +17, +18, +19, +20, +21, der(22), +der(22), +marx2 [CP10]
5	90, XX, −Y, −Y, t(8;21)(q22;q22)×2[CP10]/45, X, −X, t(8;21)(q22;q22) [10]
6	82−84, XX, −X, −X, del(1)(q21), add(1)(p36), −3, −7, −8, −9, −11, add(5)(q35), −14, −16, −18, t(8;21)(q22;q22)×2 [CP10]
7	92, XXYY, t(8;21)(q22;q22)×2 [8]/46, t(8;21)(q22;q22) [12]
8	94, XXYY, del(7)(q31)×2, +22, +22 [9]/47, XY, del(7)(q31), +22 [11]
9	83−87, XXYY, −3, −5, −7, −9, −10, −13, −13, −16, −18, i(11q)×2 [CP10]/42−43, XY, −3, −5, −13 [2]
10	1) 92, XXYY, t(15;17)(q22;q21)×2 [9]/46, XY, t(15;17)(q22;q21) [7]a 2) 46, XY,N [20]c

Notes:

a At the first diagnosis of the disease.

b At the first relapse.

c Continuous complete remission.