Evidence for a Tumor Suppressor Gene on Chromosome 19Q Associated with Human Astrocytomas, Oligodendrogliomas, and Mixed Gliomas1

(CANCER RESEARCH 52. 4277-4279, August 1. 1992| Advances in Brief

Evidence for a Tumor Suppressor Gene on Chromosome 19q Associated with Human Astrocytomas, Oligodendrogliomas, and Mixed Gliomas1

Andreas von Deimling,2 David N. Louis,3 Klaus von Ammon, Iver Petersen, Otmar D. Wiestier, and Bernd R. Seizinger Molecular Neuro-Oncology Laboratory and Neurosurgical Service, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129 [A. v. D., D. N. L,, B. R. S.J; Department of Pathology (Neuropathology), Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114 Â¡D.N. L.J; Department of Neurosurgery, University of ZÃ¼rich,Raemistrasse 100, CH-8091 ZÃ¼rich,Switzerland Â¡K.v. A.]; and Laboratory of Neuropathology, Department of Pathology, University of Zurich, Schmelzbergstrasse 12, CH-8091 ZÃ¼rich,Switzerland [L P., O. D. W.Â¡

Abstract is unique to fibrillary astrocytomas and Oligodendrogliomas, and whether a putative tumor suppressor gene locus exists on Previous studies have shown frequent allelic losses of chromosomes the long arm of chromosome 19, we evaluated 122 glial tumors 9p, 10, 17p, and 22q in glial tumors. Other researchers have briefly for LOH events on chromosome 19. reported that glial tumors may also show allelic losses of chromosome 19, suggesting a putative tumor suppressor gene locus on this chromo Materials and Methods some (D. T. Ransom et al., Proc. Am. Assoc. Cancer Res., 32: 302, 1991). To evaluate whether loss of chromosome 19 alÃelesis common in Tissue Specimens and Histopathology. Tumor and blood samples glial tumors of different types and grades, we performed Southern blot were obtained from 116 patients treated at the Massachusetts General restriction fragment length polymorphism analysis for multiple chro Hospital (Boston, MA) and at the University Hospital (Zurich, Swit mosome 19 loci in 122 gliomas from 116 patients. Twenty-nine tumors zerland). All tumors were classified by the same neuropathologists and had loss of constitutional heterozygosity of 19q, and four tumors had graded according to the guidelines of the WHO (12). The group of 122 partial deletions of 19q. Allelic losses on 19q were restricted to grade III gliomas consisted of six grade II astrocytomas, nine grade III anaplastic anaplastic astrocytomas (4/9) and grade IV glioblastomas (11/46), astrocytomas, 54 grade IV GBM, 20 pilocytic astrocytomas, five grade grade II Oligodendrogliomas (2/5) and grade III anaplastic oligoden- II Oligodendrogliomas, three grade III anaplastic Oligodendrogliomas, drogliomas (2/2), and grade II (5/8) and grade III (5/7) mixed oligoas- nine grade II oligoastrocytomas, seven grade III anaplastic oligoastro trocytomas. These data demonstrate genetic similarities between astrocytomas, two pleomorphic xanthoastrocytomas, two malignant brain- cytomas, Oligodendrogliomas, and mixed glial tumors and indicate the stem gliomas, two ependymomas, and three gangliogliomas. In four presence of a glial tumor suppressor gene on chromosome 19q. cases of GBM and one case each of anaplastic oligodendroglioma and Introduction anaplastic ependymoma, recurrent tumors were also examined. DNA Probes and RFLP Analysis. We used a panel of DNA probes LOH4 studies have been widely used to identify regions of for the long and short arms of chromosomes 19: pJCZ3.1 (D19S20) chromosomal loss in human tumors (1, 2). The loss of one and p 13-1-25 (D19S11) for the short arm; and phom 11 (D19S74), pMPSl (CYP2A), p 17.1 (D19S8), pCII-711 (APOC2), pJN2CK-M chromosomal alÃeleis thought to unmask a mutant tumor sup (CKM), pKER2 (ERCC2), pl34C (D19S51), and pEFD4.2 (D19S22) pressor gene on the remaining alÃele,thus fulfilling the para for the long arm. The loci are illustrated in Fig. 1. All probes were digm of recessively acting tumor suppressor genes (3). In glial purchased from the American Type Culture Collection. The following tumors LOH on chromosome 17p has been associated with combinations of DNA markers and restriction enzymes were used: mutations of the p53 tumor suppressor gene on the remaining PJCZ3.1 (Hinfl, Mspl, or Bglll), pl3-l-25 (Taql), phom 11 (Taql), 17p alÃele(4-6). In addition to chromosome 17p, LOH studies pMP81 (Sad), pl7.1 (Mspl), pCII-711 (Taql), pJN2CK-M (Taql), pK- in human glial tumors have revealed frequent allelic losses on ER2 (Rsal), pl34C (Pstl), and pEFD4.2 (Taql). Restriction fragment chromosomes 9p, 10, and 22q, suggesting the presence of tu length polymorphism analysis was performed as described elsewhere mor suppressor genes on these chromosomes that are involved (13). in the pathogenesis of gliomas (7, 8). Cytogenetic studies ini Results and Discussion tially suggested that loss of portions of the long arm of chro mosome 19 occur in anaplastic astrocytomas and glioblastomas The findings are summarized in Table 1. LOH for loci on 19q (9, 10). Recently, Ransom et al. (11) reported loss of chromo was demonstrated in 4 of 9 grade III anaplastic astrocytomas some 19 in 6 of 22 astrocytomas, 3 of 4 Oligodendrogliomas, (44%) and in 11 of 46 GBMs (24%), but not in the six cases of and 1 of 4 mixed oligoastrocytomas; one of their Oligodendro grade II astrocytomas. LOH was also seen in 2 of 5 grade II gliomas had a homozygous deletion at the locus D19S8 on the Oligodendrogliomas and 2 of 2 grade III anaplastic Oligoden long arm of chromosome 19 (11). To evaluate whether 19q loss drogliomas (57% of pure oligodendroglial tumors). Of the mixed oligoastrocytomas, 5 of 8 grade II tumors and 5 of 7 Received 5/20/92; accepted 6/17/92. grade III tumors had LOH (67% of mixed tumors). In four The costs of publication of this article were defrayed in part by the payment of cases (one anaplastic astrocytoma and three GBMs), partial page charges. This article must therefore be hereby marked advertisement in accord deletions were found on the long arm of chromosome 19; these ance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 Funded by Grant CD4S5 from the American Cancer Society (B. R. S.) and a are illustrated in Figs. 1 and 2. No LOH was detected in the generous gift from the Preuss Foundation (B. R. S.). other gliomas (20 pilocytic astrocytomas, two pleomorphic 2 A. v. D. is a postdoctoral fellow supported by the Deutsche Forschungsgemein schaft. To whom requests for reprints should be addressed, at Molecular Neuro- xanthoastrocytomas, two malignant brainstem gliomas, two Oncology Laboratory, Massachusetts General Hospital, Building 149, Floor 6W, ependymomas, and three gangliogliomas). LOH on the short Charlestown, MA 02129. 3 D. N. L. is a recipient of NIH Public Health Service Grant NRSA CA09144. arm of chromosome 19 was seen in only one case, a recurrent 4 The abbreviations used are: LOH, loss of heterozygosity; GBM, glioblastoma GBM in which the primary tumor did not show allelic loss on multiforme. 19p. 4277

18 262 438 420 area of homozygous deletion reported by Ransom et a/., at the D19S8 locus falls within the overlap regions of our partial deletions. Our results further demonstrate that 19q loss in glio mas appears to be restricted to diffuse astrocytic and oligoden p13.3 droglial tumors. The common region of overlap for the observed deletions p13.2p13.1 involves an area of chromosome 19 that contains a number of cloned genes, including ERCCi, ERCC2, XRCC 7, and CKM D19S11 (14, 15). The ERCC and XRCC genes are of particular interest as tumor suppressor gene candidates, since they code for DNA repair enzymes, and because ERCC2 has been recently shown to reverse the UV radiation-sensitive phenotype of xeroderma p12 pigmentosa cells (16). It remains to be shown, however, whether this locus is abnormal in glial tumors. The CKM gene, qi i iÂ»oft/ encoding muscle creatine kinase, could also be involved in a tumor suppressor pathway, since recent studies have shown that q12q13.1qi3.2q13.3q13.4u the upstream enhancer region of the murine CKM gene con tains a locus which can be activated by the murine p53 tumor suppressor gene product (17). Numerous studies have impli cated ihe p53 gene in glial tumorigenesis (4-6, 18, 19). If CKM CYP2A/.^ plays a role in a cellular pathway involving p53, perhaps in control of the cell cycle, mutations in the CKM gene itself may D19S8^^ be important in oncogenesis. In these four cases, however, rel atively large areas of 19q were not informative or not examined CKM^^^ (Fig. 1, shaded areas), and we cannot therefore exclude other ERCC2 regions of 19q as sites for this putative tumor suppressor gene. ~~D19S51D19b22Studies of allelic loss have shown that certain chromosomal losses are more common in some tumors than in others and that patterns of allelic loss may characterize individual tumors. Loss of chromosome 19 is an uncommon finding in human Fig. 1. Schematic of chromosome 19. Four cases with partial deletions on tumors (20) and has been reported, to our knowledge, only in chromosome 19. Deleted areas, LOH; shaded areas, not informative or not ex glial tumors (11). The histogenesis of mixed oligoastrocytomas amined; white areas, maintenance of heterozygosity. has long been a source of debate. Clinical, immunohistochem- ical, electron microscopic, and experimental data have hinted Table 1 Tumor types and number of tumors assessed that the oligodendroglial and astrocytic components in mixed oligoastrocytomas may arise from a common stem cell and that DiagnosisPilocytic (patients)"206954 19p*0/170/60/91/350/50/20/80/40/20/10/21/91LOH19q*0/100/64/9 mixed tumors are related to "pure" oligodendrogliomas (21- astrocytomaWHO 23). The common loss of portions of the long arm of chromo IAstrocytomagrade IIAnaplastic WHO grade some 19 in mixed oligoastrocytomas, oligodendrogliomas, and astrocytomaWHO (44%)11/46(24%)2/5 IIIGlioblastomagrade multiformeW (50)5397222 APOC2 CKM IVOligodendrogliomaHO grade IIAnaplastie WHO grade (40%)2/2(100%)5/8 oligodcndrogliomaWHO IIIOligoastrocytomagrade â€¢¿^4.5 <-4.4 IIAnaplastic WHO grade (63%)5/7 OligoastrocytomaWHO (71%)0/20/20/10/229/100 IIIMalignantgrade 3.8 3.5 gliomaPleomorphicbrain stem xanthoastrocytomaAnaplastic ependymomaWHO (1)3 <- 2.2 IIIGangliogliomaTotalTumorsgrade <-2.0 (2)122(116)LOH

(29%) â€¢¿Whenmultiple tumors from the same patients are included, the number of 4.7 -> individual patients is listed in parentheses. * Numerator represents the number of tumors with LOH; denominator repre 2.0 sents the number of informative patients. 1.8 3.1

Loss of allelic heterozygosity on the long arm of chromosome 19 is therefore a frequent event in glial tumorigenesis and sug gests that a tumor suppressor gene on 19q is involved in the D19S51 D19S22 pathogenesis of astrocytic and oligodendroglial tumors. Our Fig. 2. Interstitial deletion in an anaplastie astrocytoma (case 262). Left, ref results are in agreement with the findings of Ransom et al. who erence DNA; right, tumor DNA. LOH is seen in APOC2, CKM, and DI9S5I; heterozygosity is maintained in D19S22. With the CKM probe only the upper showed allelic losses of chromosome 19q in astrocytomas, polymorphism is informative; the hybridization signal in the 2.2-kilobase band is mixed gliomas, and oligodendrogliomas (11). In addition, the reduced in the tumor tissue. Numbers, band sizes in kilobases. 4278

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Downloaded from cancerres.aacrjournals.org on September 23, 2021. © 1992 American Association for Cancer Research. Evidence for a Tumor Suppressor Gene on Chromosome 19q Associated with Human Astrocytomas, Oligodendrogliomas, and Mixed Gliomas

Andreas von Deimling, David N. Louis, Klaus von Ammon, et al.

Cancer Res 1992;52:4277-4279.

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