Chromosome 5 Abnormalities in Acute Lymphoblastic Leukemia

Francisco Palau, Felix Prieto, Lourdes Badia, Magdalena Beneyto, and Isana Benet

ABSTRACT: We report two cases of acute lymphoblastic leukemia with involvement of chromosome 5. One of them showed a del(5)(q13q33) in a 5-year-old boy who had previously received antineoplastic chemotherapy for an L1-ALL that had been diagnosed nine months before. '/'he other one showed a t(5;7)(q12-13;q36) together with a t(8;14)(q24;q32) and a der( l ) in a 66-year- old man with an L3-ALL. Both chromosome 5 aberrations are interpreted as evolutionary events. In the first case, it was secondary to chemotherapy treatment; in the second, an evolutionary chromosome rearrangement, considering the translocation between chromosomes 8 anti 14 as the primary cytogenetie event.

INTRODUCTION

The most frequent and specific clonal cbromosomal abnormali t ies found in acute lymphoblas t ic leukemia (ALL) are t(1:9)[q23;p13), t(4:l 1)(q21 ;q23), t(9;22)(q34:ql 1), t(8;14)(q24;q32), t[11;14)(p13;q13), and delet ion of the long arm of chromosome 6 [1-3]. Chromosomal aberrat ions involving chromosome 5 are frequent in lnyelodys- plastic syndromes and acute myeloid leukemia, often as the 5 q - anomaly [4, 5]. but are rare in ALL and other lymphoid neoplasms [5, 6].

We report two ALL cases with part ic ipat ion of the 5q12-13 band in their respective chromosomal abnormali t ies. We consider the possibi l i ty of these being evolut ionary aberrations, one of them secondary to previous ant ineoplast ic chemotherapy and the other as an evolut ionary clonal event.

MATERIALS AND METHODS

Cytogenetic Investigations

Chromosome studies were performed on bone marrow cells that were processed in both cases by a direct method and 48-hour nonst imulated culture in RPMI 1640 with 15% fetal calf serum. Case 2 bone marrow cells were also processed in a 48- hou r - s t imu la t ed culture using 12-O-tetradecanoylphorbol 13-a(:etate (TPA) (2 /xg/ mL) as a polyclonal B-cell activator and low-molecular-weight B-cell growth factor

From U n i d a d de Gen6tica, Hospi ta l La Fe (F. P.. F. P.. L. B., M. B.] and Servicio de l l emato log ia y Oncologia , l lospital Clinico, Un ive r s idad de Valencia (I. B.), Valencia, Spain.

Address reprint requests to: Dr. F. Palau, Department of Biochemistry & Molecular Genetics, St. Mary's Hospital Medical School, Norfolk Place, London W2 1PG, United Kingdom.

Received November 29, 1989: accepted May 11, 1990.

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1 7 4 F. Palau et al.

(BCGF-I or IL-4) 10%, v/v, or inter leukin 2 (IL-2) 10%, v/v, (Cellular Products, Buffalo) as B-cel l -prol i fera t ing factors, in order to obtain more dividing ceils.

Chromosomes were character ized by a trypsin-G banding method [7]. Chromo- somal abnormal i t ies are descr ibed according to the ISCN [8].

Morphologic and Immunologic Analysis

The diagnosis of acute lymphoblas t ic leukemia was based on morphologic and tyro- chemical s tudies of the per iphera l blood smears and bone marrow aspirates prior to therapy, according to the French-American-Bri t ish (FAB) Cooperative Group criteria [9].

Cell-surface antigens were detected by a s tandard indirect immunofluorescent assay with monoclona[ ant ibodies representat ive of the major cluster groups (CD). They inc lude J5 (CALLA or CD10), HLA-DR, B-cell and B-cell precursor-associated antigens B4 (CD19), B1 (CD20) and BA-2 (CD9), T-ce l l -associa ted antigens Leu-9 (CD7), O K T l l (CD2) and OKT3 (CD3), and myeloid-associa ted antigen My-9 (CD33). Cell-surface immunoglobul in chains were detected by direct fluorescence with fluo- resceinated goat ant i -human immunoglobul in chains. Cytoplasmic ~-chain immuno- globul in (C/~) was identif ied by a direct method that used fluoresceinated F(ab)~ goat ant i -human/~-chains .

RESULTS

Clinical and Morphologic Features

Patient No. 1. A 4-year-old boy was admit ted to our hospital in March 1988, because fifteen days earl ier he showed polyuria, polydipsia , vomiting, pain in the calves making walking difficult, and hematomas. He had had a history of anorexia, weight loss, and general ized weakness for six months. On physical examinat ion, he had some petechiae on the neck and thorax, laterocervical adenopathies , a systolic murmur, and hepatomegaly 3 cm below the rib cage. Peripheral blood showed 17.5 × 109/L white blood cell (WBC) count with 40% blast cells, the hemoglobin was 85 g/dL, and the platelet count was 82 x 10~VL. The bone marrow revealed a monomorphic infil- trate of 90% blasts. The blast cell morphology was consistent with acute lymphoblas t ic leukemia. The cytochemical staining reactions were negative for Sudan black B, myeloperoxidase , per iodic acid Schiff, beta-glucuronidase and acid phosphatase, and posit ive for a lpha-napthyl acetate esterase (81%). Immunologic data are shown in Table 1. A diagnosis of L1-ALL with common immunopheno type was made. The boy received specific chemotherapy (etoposide, cytarabine, vincrist ine, L-aparaginase, prednisone, and intrathecal prophylaxis) and complete remission was obtained. Dur- ing the induct ion chemotherapy, he underwent a convulsive episode secondary to a parietal hemorrage that was at tr ibuted to L-asparaginase coagulopathy. Nine months later, he had a hematologic relapse with 91% blast cells in the bone marrow aspirate. The WBC count was 6.1 × 109 with 9% blast cells, the hemoglobin level 86 g/L, and the platelet count 57 x 1 0 9. The spinal fluid was normal. The morphology was L1 by FAB criteria. Immunopheno type is shown in Table 1.

Patient No. 2. A 66-year-old mall was admit ted in March 1989. The patient had all history of fatigue, weight loss, and profuse sweating. The physical examinat ion was normal. On admission, the WBC count was 9.11 x 109/L with 9% blast cells and platelet count 252 x 109/L. The bone marrow specimen showed an infiltrate of

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Table 2 Cytogenet ic analysis of pat ients wi th ALL and a chromosome 5 abnormal i ty

Patient Clilfi(:al Number of cells Number of cells number stage Sour(:e" analyzed abnormal Karyotype

1 Diagnosis Relapse

2 l)iagnosis

BM 48 hr 9 0 46,XY BM 48 hr 6 2 46.XY,dcl(5)(q13(133) BM 48 hr 2 2 4(LXY,t(5;V)(ql 3:q36).

BM TPA + l(8;14)(q24;q32) (74%),' t3CGF-I 1(} 10 46,XY,t(5:7)(q13:q36),

BM TPA t t(8:14)(q24;q32), der(1} IL-2 7 7 ( l l } l e r ~ { : e n --,lq31 : :1q23

I {th,*r ) (26%1

"BM. bone marrow: 48 hr, cells cultured for 48 hours in the absence of mitogens: TPA. 12-Oqetrad(x:anoylphorbo113- acetate; BCGF-I, B-cell growth factor type I or interleukin 4; ]L-2. interleukin 2.

l y m p h o i d blasts w i t h a basoph i l i c scarce cy toplasm, cop ious vacuoles , and nucleol i . An L3-ALL by the FAB class i f icat ion was diagnosed. There was a blast inf i l t ra t ion of the sp ina l fluid. The pat ient d ied three days later.

Cytogenetic Analys i s

The resul ts of cy togene t ic inves t iga t ions of the l eukemia cells f rom the two pat ients are l i s ted in Table 2. Pat ient no. 1 was s tud ied at d iagnosis showing a d ip lo id karyotype; at re lapse n ine m o n t h s later, w h e n he was under c h e m o t h e r a p y t reatment , a del(5)(q13q33) was found in two of the six (;ells ana lyzed {Fig. 1). Patient no. 2 was s tud ied at d iagnosis s h o w i n g a typical t(8;14)(q24;q32), usual ly found in Burkit t - l ike (Jells of L3-ALL, and a t(5:7)(q12 13:q36) in tim 19 (:ells ana lyzed (Fig. 2). Moreover , a de r iva t ive c h r o m o s o m e de r ( l ) t ( lp t e r - - , cen - - ,1q31 : : lq23--~1qter) was found in a subc lone (26.3% of cells). In this case, we obta ined more adequa te me taphases in the s t imula ted (:ultures. Also, tim oon- s t imu la t ed short cu l ture showe(t two metaphases wi th the same clonal karyotype.

DISCUSSION

The 5 q - a n o m a l y and o ther c h r o m o s o m a l aberrat ions i nvo lv ing c h r o m o s o m e 5 are rare f indings in l y m p h o i d neoplasms . A m o n g the 234 cases of ALL wi th s t ructura l rea r rangements s tud ied in the Thi rd In ternat ional Workshop on C h r o m o s o m e s in Leukemia , c h r o m o s o m e 5 was translocate(t or de le ted in only three cases [61. In their r ev iew of 208 d isorders associa ted wi th 5(t ( :hromosome, Van Den Bergbe et al. 15] found on ly n ine l ymphopro l i f e r a t i ve disorders , three of them ALL in chi ldren . In fact, the 5 q - anomaly is associa ted wi th mye lodysp la s t i c synd romes and acute myelo id l eukemia as de novo or se(:ondary d isorders [5 ,10 121 . Thus , our two patients show a rare morpho log ic -cy togene t i c cmnbina t io lL

Pat ient no. 1 s h o w e d a l y m p h o i d c lone wi th a 5q ch romosome , w h i c h appea red at re lapse n ine nmnths after d iagnos is of an ALL wi th a normal karyotype. S ince then, the pat ient has rece ived an t ineop las t i c chemothe rapy . This is not a f requent event . A m o n g the 21 ALL cases s tud ied at re lapse in the Sixth In ternat ional Workshop on C h r o m o s o m e s in Leukemia , only one case wi th a normal karyotype at d iagnosis s h o w e d an abnorma l c lona l evo lu t ion [13]. Thus, we cannot de t e rmine in our pat ient whe the r there has been a t rue relapse wi th clonal evo lu t ion in a c h r o m o s o m a l l y normal l eukemic c lone or whe the r it represents a secondary l eukemia event . The

Chromosome 5 Abnorlnalities in ALL 177

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Figure 1 Trypsin-Giemsa-banded karyotype showing interstitial deletion on the long arm of chromosome 5, del(5)(q13q33) (patient no. 1).

finding of a 5 q - chromosome suggests the second possibility as more probable. Cytostatic treatment was probably the cause of the relapse, although the cell pheno- type in secondary leukemia is usually myeloid with loss of chromosomes 5 or 7 [10, 11]. We found only two clonal cells, though the presence of normal cells is frequent in 5 q - anomaly [5]. In this patient, blast cells showed at immunologic data diagnosis suggesting a precursor B-cell origin with negativity for the myelomonocytic antigen CD33 and also for myeloperoxidase and Sudan black B. The presence of a high number of alpha-napthyl acetate esterase-pnsitive (:ells could suggest the diagnosis of a bich)- hal hybrid leukemia [14]. This is a possibility because there is no previous chemother- apy which could have induced a secondary acute leukemia or a switch in cell lineage [15]. Then, at relapse, only the lymphoid line would have suffered a clonal evolution. In spite of these arguments, we think that negativity for the CD33 antigen and the presence of specific B-cell antigens suggests more a lymphoid origin than a biclonality. The presence of non-specific esterase activity in leukemic cells could be the conse- quence of a transient phase of limited promiscuity of gene expression, which remains as a relic in leukemic blast cells that are in maturation arrest [16].

Patient no. 2 showed a L3-ALL with a t{8;14)(q24;q32), typically described in these leukemias and in Burkitt 's lymphoma [1]. He also showed a t(5;7)(q12 13;q36), which is a rare chromosomal aberration in ALL. The 5q12-14 breakpoint is frequently affected in the interstitial deletion of 5 q - anomaly [5].

Some chromosomal aberrations affecting chromosome 5 have been des(:ribed in ALL as the t(5;14)(q31:32) associated with hypereosinophil ia [17.18] and inversion of chromosome 5 affecting the 5ql 3 band 119]. They are different structural abnormalities than that seen in our case.

178 F. Palau et al.

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3 4 5 der5

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Figure 2 Trypsin-Giemsa-banded karyotype showing the t(5;7) (q12 - 13;q36) together with the classical t(8;14}(q24;q32) (patient no. 2).

Raimondi et al. [20] have described a group of ALL with involvement of 7q32-36 region, where the T-cell receptor beta chain gene is located. These leukemias showed a T-cell lineage that is different from the B-cell immunophenotype showed by our patient no. 2.

Translocations between chromosomes 5 and 7 have been reported in three malig- nant myeloid disorders involving the 5q11.2 snbband, but with the participation of the short arm of chromosome 7 [21]. Obviously, the cellular origin and the type of translocation are different from that in our case.

We conclude that ALL can be associated with chromosome 5 abnormalities but they are a rare finding in this type of leukemia. In our cases, we consider these abnormalit ies as evolutionary chromosomal events, in case no. 1 as secondary to chemotherapy and in case no. 2 as an evolutionary abnormality in the biology of the neoplasia, probably giving the leukemia cells a proliferative advantage.

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