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Genetic heterogeneity and clonal evolution in neuroblastoma

Abstract

Tumour heterogeneity and clonal evolution at the genetic level may explain the development of malignant or resistant disease during clinical progression of neuroblastoma (NB). In this report we use 1p allelic analysis and DNA ploidy to evaluate clonal heterogeneity and clonal selection in vivo. We studied a total of 69 tumours from 29 patients with NB. To evaluate tumour heterogeneity and clonal evolution in vivo we used a panel of polymorphic allelic markers mapping to chromosome 1. 33 tumours from 12 patients (group 1) were obtained from different sites during the same surgery or at sequential surgeries without intervening chemotherapy to evaluate genetic heterogeneity. Paired samples from 10 patients (group 2) were used to evaluate clonal selection before and after chemotherapy. In 6 cases paired tumours and derived cell lines were studied. Analysis of DNA ploidy changes by karyotype, FISH and flow cytometry was performed in 15 tumours from 6 multiply recurred local-regional (LR) NB patients. Allelotype study revealed that 66% (8/12) of group 1 samples were heterogeneous, with distinct allelic patterns in tumour samples separated by time or location. In group 2 allelic patterns were different in post-chemotherapy specimens in 60% (6/10). DNA ploidy analysis showed that pre-chemotherapy samples contained 2 distinct ploidy clones, one diploid and one triploid, whereas all post-chemotherapy tumor samples were 100% diploid. These findings suggest that NB exhibits a high degree of clonal heterogeneity and clonal evolution occurs during the course of therapy and clinical progression. © 2001 Cancer Research Campaign http://www.bjcancer.com

References

  1. Ambros PF, Ambros IM, Strehl S, Bauer S, Luegmayr A, Kovar H, Ladenstein R, Fink FM, Horcher E, Printz G, Mutz I, Schilling F, Urban C and Gadner H (1995) Regression and progression in neuroblastoma. Does genetics predict tumor behaviour?. Eur J Cancer 31A: 510–515

  2. Ambros IM, Zellner A, Roald B, Amann G, Ladenstein R, Printz D, Gadner H and Ambros PF (1996) Role of ploidy, chromosome 1p, and Schwann cells in the maturation of neuroblastoma. N Engl J Med 334: 1505–1511

  3. Brodeur GM (1995) Molecular basis for heterogeneity in human neuroblastomas. Eur J Cancer 31A: 505–510

  4. Brodeur GM, Hayes FA, Green AA, Casper JT, Wasson J, Wallach S and Seeger RC (1987) Consistent N-myc copy number in simultaneous or consecutive neuroblastoma samples from 60 individual patients. Cancer Res 47: 4248–4253

  5. Brodeur GM, Pritchard J, Berthold F, Carlsen NLT, Castel V, Castleberry RP, De Bernardi B, Evans AE, Favrot M, Hedborg F, Kaneko M, Kemshead J, Lee REJ, Look T, Pearson ADJ, Philip T, Roald B, Sawada T, Seeger RC, Tsuchida Y and Voute PA (1993) Revisions of the International Criteria for neuroblastoma diagnosis, staging, and response to treatment. J Clin Oncol 11: 1466–1477

  6. Brodeur GM, Sawada T, Tsuchida Y and Voute PA (1999). Neuroblastoma, Elsevier Science: Amsterdam

  7. Cawkwell L, Bell SM, Lewis FA, Dixon MF, Taylor GR and Quirke P (1993) Rapid detection of allele loss in colorectal tumours using microsatellites and fluorescent DNA technology. Br J Cancer 67: 1262–1267

  8. Cheung NKV, Kushner BH, LaQuaglia M, Kramer K, Gollamudi S, Heller G, Gerald W, Yeh S, Finn R, Larson SM, Wuest D, Byrnes M, Dantis E, Mora J, Cheung IY, Rosenfield N, Abramson S and O’Reilly RJ (2001) N7 a novel multi-modality therapy of high risk neuroblastoma in children diagnosed over 1 year of age. Med and Ped Oncol 36: 227–230

  9. Fearon ER and Vogelstein B (1990) A genetic model for colorectal tumorigenesis. Cell 61: 759–767

  10. Gotoh T, Sugihara H, Matsumura T, Katsura K, Takamatsu T and Sawada T (1998) Human neuroblastoma demonstrating clonal evolution in vivo. Genes Chromosomes Cancer 22: 42–49

  11. Hedley DW (1989) Flow cytometry using paraffin-embedded tissue: five years on. Cytometry 10: 229–241

  12. Kushner BH, LaQuaglia MP, Bonilla MA, Lindsley K, Rosenfield N, Yeh S, Eddy J, Gerald WL, Heller G and Cheung NKV (1994) Highly effective induction therapy for stage 4 neuroblastoma in children over 1 year of age. J Clin Oncol 12: 2607–2613

  13. Larson AA, Kern S, Curtiss S, Gordon R, Cavenee WK and Hampton GM (1997) High resolution analysis of chromosome 3p alterations in cervical carcinoma. Cancer Res 57: 4082–4090

  14. Mao L, Lee DJ, Tockman MS, Erozan YS, Askin F and Sidransky D (1994) Microsatellite alterations as clonal markers for the detection of human cancer. Proc Natl Acad Sci 91: 9871–9875

  15. Martinnsson T, Sjoberg RM and Hedborg F (1995) Deletion of chromosome 1p loci and microsatellite instability in neuroblastomas analyzed with short-tandem repeat polymorphisms. Cancer Res 55: 5681–5686

  16. Mora J, Cheung NKV, Kushner BH, LaQuaglia MP, Kramer K, Fazzari M, Heller G, Chen L and Gerald W (2000) Clinical Categories of Neuroblastoma are associated with different Patterns of Loss of Heterozygosity on Chromosome Arm 1p. J Mol Diagn 2: 37–46

  17. Mora J, Gerald W, Chen L, Qin J and Cheung NKV (2001) Survival analysis of clinical, pathologic and genetic features in neuroblastoma presenting as local-regional disease. Cancer 91: 435–442

  18. Shimada H, Chatten J, Mewton WA, Sachs N, Hamoudi AB, Chiba T, Marsden HB and Misugi K (1984) Histopathologic prognostic factors in neuroblastic tumors: definition of subtypes of ganglioneuroblastoma and an age-linked classification of neuroblastomas. J Natl Cancer Inst 73: 405–416

  19. Shimada H, Ambros IM, Dehner LP, Hata J, Joshi VV and Roald B (1999) Terminology and morphologic criteria of neuroblastic tumors: recommendations by the International Neuroblastoma Pathology Committee. Cancer 86: 349–363

  20. Suzuki T, Mugishima H, Chin M, Takamura M, Shichino H, Nagata T and Harada K (1997) Case of neuroblastoma with differing cytologic and molecular biologic features at primary and metastatic sites. J Ped Hem/Onc 19: 176–180

  21. Taylor SR, Blatt J, Constatino JP, Roederer M and Murphy RF (1988) Flow cytometric DNA analysis of neuroblastoma and ganglioneuroma. Cancer 62: 749–754

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Correspondence to J Mora.

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Keywords

  • heterogeneity
  • neuroblastoma
  • clonal evolution
  • DNA ploidy

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