The cytogenetics of Wilms' tumor

The Cytogenetics of Wilms' Tumor Rosalyn M. Slater

ABSTRACT: A close association has been demonstrated between the congenital deletion 11p13 and predisposition to Wilms' tumor. Recent cytogenetic studies on Wilms' tumor cells from normal children strongly suggests that somatic changes in the short arm of chromosome #11 play an important role in the development of this tumor. The application of improved cytogenetic techniques coupled with molecular biologic analysis may help resolve questions regarding the requirement of additional changes (to alterations of 11p13) in order to evoke complete transformation leading to malignancy. INTRODUCTION Wilms' tumor of the kidney is the most c o m m o n abdominal solid tumor of childhood, occurring at a frequency of about 1 in 10,000 young children. It is frequently associated with a n u m b e r of congenital abnormalities, such as sporadic aniridia, hemihypertrophy, and genitourinary anomalies [1]. Children with aniridia have a 1-in-3 chance of developing Wilms' tumor, and the a n i r i d i a - W i l m s ' tumor association has been s h o w n to be linked to a deletion of the short arm of chromosome #11, [del(11)(p13)] [2, 3]. Based on the incidence of familial and nonfamilial cases, Knudson and Strong [4] have proposed that two mutations are necessary for the development of Wilms' tumor; it has been suggested that the 11p deletion may be one of the events [5]. If this proposal is correct, then a similar somatic change should be present in the Wilms' tumor cells from normal individuals. Recent reports using cytogenetic and molecular biological techniques have added support to this theory. CYTOGENETIC STUDIES ON WILMS' TUMOR CELLS Until 1981, cytogenetic investigations on Wilms' tumors from normal children had been limited to one report using c o n v e n t i o n a l staining techniques [6]. Subsequently, data has become available on 38 tumors studied after short-term culture techniques [7-14] and on one cell line [15] (Table 1). With the exception of one case [10], the tumor material was obtained from children with no congenital chromosome abnormalities as verified in the majority of cases either by analysis of stimulated peripheral blood lymphocytes or by the presence of cells with an apparently normal karyotype mixed with the t u m o r cell population. A total of 13 tumors were found to show variation in the short arm of chromosome #11 as a result of a simple From the Institute of Human Genetics, Universityof Amsterdam,AcademicMedical Center. Subfaculty of Medicine,Amsterdam, The Netherlands. Address requests for reprints to Dr. Rosalyn M. Slater, Institute of Human Genetics, University of Amsterdam, Academic Medical Center, Subfaculty of Medicine, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Received August 5, 1985; accepted August 19, 1985.

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Cancer Genet Cytogenet19:3741 (1986) 0165-4608/86/$03.50

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R . M . Slater

deletion or due to translocations with other chromosomes. A d d i t i o n a l l y , a child with a constitutional 11p deletion s h o w e d basically the same karyotype in his Wilms' tumor [10]. Cytogenetic studies on a s p e c i m e n of n o d u l a r renal blastema, a possible precursor lesion of W i l m s ' tumor, from a patient with concomitant Wilms' also showed the presence of an interstitial del(11)(p11-p14) in cultured cells [16]. The remainder demonstrated no apparent changes in this region of #11 w i t h i n the limits of resolution of the cytogenetic techniques e m p l o y e d . The next most c o m m o n change involved variation in the long arm of c h r o m o s o m e #1. Kondo et al. [101 reported five of eight cases with partial trisomy for 1q21-1q31. These i n c l u d e d three patients where this change was a c c o m p a n i e d by partial m o n o s o m y of the long arm of #16, either due to der(16),t(1;16) (initially reported by Kaneko et al. [11]) or the result of an unidentifiable translocation together with an i s o c h r o m o s o m e lq. Douglass et al. [14] also found variation in l q in five cases and del(1)(p13) in two patients. Closer e x a m i n a t i o n of these data [14] also reveals that four s h o w e d monosomy of 16q either due to a translocation or simple nondisjunction, three of which also had l q variation in the same cell. The results of Slater et al. [8, 13] showed structural variation in #1 in three cases; changes in # 3 being more common, present in four cases. This latter finding, however, was not substantiated by other investigators. In those tumors reported as having h y p e r d i p l o i d variation, an extra chromosome # 9 and an extra #12 were the most consistent n u m e r i c a l abnormalities [11, 12, 14]. Structural aberrations involving the short arm of c h r o m o s o m e #11, therefore, appear to be the most c o m m o n a b n o r m a l i t y in W i l m s ' t u m o r cells. Variation in lq has been reported for a n u m b e r of different types of h u m a n malignancy, and appears to be associated with tumor progression [17]. However, abnormalities of chromosome #16 are very rare in t u m o r cells, though they have been observed in retinoblastoma, in some instances in association with aberrations of chromosome #1 [18, 19]. This suggests that these changes involving #1 and # 1 6 m a y p l a y a role in the d e v e l o p m e n t of both types of e m b r y o n i c tumor. A c o m p a r i s o n of the aforementioned data on W i l m s ' tumor reveals that there may be a relationship between t u m o r histology and c h r o m o s o m e number. The majority of the tumors studied have h a d c h r o m o s o m e numbers w i t h i n the diploid range. Three tumors were found to be h y p e r t r i p l o i d and two were h y p o d i p l o i d with h y p e r t r i p l o i d sublines due to p o l y p l o i d i z a t i o n of the m a i n stem line [13, 14]. All but one of these tumors showing h y p e r t r i p l o i d variation were reported as having an anaplastic histology, w h i c h has not yet been associated with tumors in the dipl o i d - h y p e r d i p l o i d range. POSSIBLE ROLE OF 11p IN DEVELOPMENT OF WILMS' TUMOR

The close association of the congenital deletion 11p13 w i t h a p r e d i s p o s i t i o n to W i l m s ' tumor and the results of cytogenetic studies on W i l m s ' t u m o r cells from normal children strongly suggests that somatic changes in the short arm of chromosome #11 play an important role in the d e v e l o p m e n t of this tumor. The close association of the breakpoints involved in c h r o m o s o m e rearrangements in cancer cells with the localization of h u m a n cellular protooncogenes as found with c°abl in chronic m y e l o i d l e u k e m i a (CML) with t(9;22) [20] and c-myc in Burkitt's lyrnp h o m a with t(8;14) and variant translocations [21], has led to speculation about the possible role of the Harvey ras protooncogene (c-Ha-ras 1) in W i l m s ' tumor because of its location on the short arm of #11 [22]. Studies on seven patients with 11p deletions and the a n i r i d i a - W i l m s ' tumor s y n d r o m e have s h o w n that this oncogene is neither located in the deleted segment [23-25], nor suffers any gross rearrangement because of the deletion [25].

39

Cytogenetics of Wilms' T u m o r

Table 1 Cytogenetic studies on W i l m s ' t u m o r Cytogenetic findings Number studied

Numerical range

Structural variation in 11p

Other variations

Ref.

TUMOR 1 2

46 46

1 2

4 1 2

46-52 46 46(51)

3° 1 --

5

46-54

--

14

38-78

6

9

46-77

1

1

46

1

1

not given

1

none seen 4p + ,del(9)(q22) 4p+,#7,2 + 3 , + 6 . + 7 ,+ 8 ,+ 9 ,÷ 12 t(11;12)(p13;q13) - 16, + der(16)t(1;16](q21?;q22?) - 16, + der(16)t(1;16)(q11?;q11?) del(16)(q12 - q22?),:(1q) lq abnormalities + 9, + 12 #1 #16q (4 cases, 3 with lq variation) +9 and/or + 12 #3q #14 #1 CELL LINE + 19 RENALNODULE not given

[7] [8] (1 case)

[10] [9] [11]

(1 (2 (3 (7

[12]

case) cases) cases) cases)

(6 cases) (4 cases) (3 cases) (3 cases)

[14]

[13]

[15] [16]

~Onepatient with constitutional del(11)(p13).

A comparison of constitutional and t u m o r DNA from Wilms' t u m o r patients using a n u m b e r of DNA probes for the short arm of c h r o m o s o m e #11, including cHa-ras 1, has s h o w n that in some cases heterozygosity in the constitutional DNA has been replaced by homozygosity for the 11p markers in the t u m o r cells and, thus, provide e v i d e n c e that mitotic events have occurred w h i c h could serve to unmask initial predisposing recessive mutations [26-29]. Similar results using DNA probes for # 1 3 had also previously been obtained for retinoblastoma, another embryonic t u m o r of c h i l d h o o d [30]. In certain cases of Wilms' tumor, one of the two constitutional c-Ha-ras 1 alleles was absent, w h i c h suggests that changes in this oncogene may play some role in the d e v e l o p m e n t of this tumor [28]. Karyotype analysis was only carried out in four [26,28] of the a b o v e m e n t i o n e d Wilms' tumors and structural variation 11p could be detected in one case [28], demonstrating that changes in 11p are probably more frequent in W i l m s ' tumor than is evident from cytogenetic studies alone. These findings suggest that a recessive mutation carried on the short arm of #11 plays an important role in the d e v e l o p m e n t of W i l m s ' tumor. The fact that not all children with constitutional del(11)(p13) d e v el o p W i l m s ' t u m o r is e v i d e n c e that either additional genetic changes are required for complete transformation to the malignant p h e n o t y p e or that small, as yet undetected, differences exist in the size of the deletion. The application of i m p r o v e d cytogenetic techniques and a molecular approach may help to resolve this problem.

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R . M . Slater

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25. Geurts van Kessel A, Nusse R, Slater R, Tetteroo P, Hagemeijer A (1985): Localization of the oncogene c-Ha-ras 1 outside the aniridia-Wilms' tumor-associated deletion of chro~ mosome 11 (delllp13) using somatic cell hybrids. Cancer Genet Cytogenet 15:79-84. 26. Koufos A, Hansen MF, Lampkin BC, Workman ML, Copeland NG, Jenkins NA, Cavenee WK (1984): Loss of alleles at loci on h u m a n chromosome 11 during genesis of Wilms' tumor. Nature 309:170-172. 27. Orkin SH, Goldman DS, Sallan SE (1984): Development of homozygosity for chromosome 11p markers iri Wilms' tumour. Nature 309:172-174. 28. Reeve AE, Housiaux PJ, Gardner RJM, Chewing WE, Grindley RM, Millow LJ (1984): Loss of a Harvey ras allele in sporadic Wilms' tumor. Nature 309:174-176. 29. Fearon ER, Vogelstein B, Feinberg AP (1984): Somatic deletion and duplication of genes on chromosome 11 in Wilms' tumours. Nature 309:176-178. 30. Cavenee WK, Dryja TP, Phillips RA, Benedict WF, Godbout R, Gallie BL, Murphree AL, Strong LC, White RL (1983): Expression of recessive alleles by chromosomal mechanisms in retinoblastoma. Nature 305:779-784.