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Extensive Intra-tumor Heterogeneity in Primary Human Glial Tumors as a Result of Locus Non-specific Genomic Alterations

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Abstract

Genomic changes are a hallmark of the neoplastic process. These range from alterations at specific loci and defined karyotypic changes which influence tumor behavior to generalized alterations exemplified by microsatellite instability. Generalized genomic changes within a tumor would be evidence in favor of the mutator hypothesis which postulates a role for such extensive changes during tumorigenesis. In this report, we have used the DNA fingerprinting technique of randomly amplified polymorphic DNA (RAPD) analysis to study genomic alterations within primary human astrocytic tumors (gliomas) in a locus non-specific manner. The RAPD fingerprinting profile of consecutive segments of tumors 2 mm across was studied; 17 astrocytic (high- and low-grade) tumors were sectioned end to end. Tissue from 50 consecutive sections, 40 µm thick (total 2 mm across), was pooled and taken to be a tumor compartment. DNA was subjected to RAPD amplification by 15 random 10-mer primers.

A tumor segment was taken to have a DNA fingerprinting pattern different from others in the same specimen when its RAPD profile differed from others by at least one band of one RAPD reaction. All but one of the tumors showed compartments with a unique genetic profile, indicating genomic instability leading to widespread intra-tumor genetic heterogeneity. Eight tumors were also studied for loss of heterozygosity (LOH) of the p53 and D17S379 loci in the different segments as examples of alteration of specific tumor influencing loci. Three showed LOH of p53, which was limited to only one compartment of each tumor.

The extensive intra-tumor genetic instability detected in this study is suggestive of the overall high rate of change in the genomes of tumors including those of a lower grade. It is hypothesized that some of these altered clones, which manifest as zones of heterogeneity in a solid tumor, may accumulate changes at loci known to influence tumor behavior, and thus clinical outcome.

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References

  1. Heppner GH: Tumor heterogeneity. Cancer Res 44: 2259-2265, 1984

    Google Scholar 

  2. Barranco SC, Drewinko B, Humphrey RM: Differential response by human melanoma cells to 1,3-bis-(1-nitrosourea) and bleomycin. Mutat Res 19: 277-280, 1973

    Google Scholar 

  3. Calabresi P, Dexter DL, Heppner GH: Clinical and pharmacological implications of cancer cell differentiation and heterogeneity. Biochem Pharmacol 28: 1933-1941, 1979

    Google Scholar 

  4. Edwards PA: Heterogeneous expression of cell surface antigens in normal epithelia and their tumors revealed by monoclonal antibodies. Br J Cancer 51: 149-160, 1985

    Google Scholar 

  5. Manson MM, Green JA, Neal GE: Monoclonal antibody against tumor derived cell line, shows heterogeneity of altered hepatocytes during AFB-induced carcinogenesis. Int J Cancer 34: 869-874, 1984

    Google Scholar 

  6. Sinha S, Marshall C, Neal GE: Gamma-glutamyltranspeptidase and the ras-induced transformation of a rat liver cell line. Cancer Res 46: 1440-1445, 1986

    Google Scholar 

  7. Dexter DL, Kowalski HM, Blazar BA, Fligiel Z, Vogel R, Heppner GH: Heterogeneity of tumor cells from a single mouse mammary tumor. Cancer Res 38: 3174-3181, 1978

    Google Scholar 

  8. Mitelman F, Mark J, Levan G, Levan A: Tumor etiology and chromosome pattern. Science 176: 1340-1341, 1972

    Google Scholar 

  9. Mitelman F: The chromosomes of fifty primary Rous rat sarcomas. Hereditas 69: 155-186, 1971

    Google Scholar 

  10. Wright S: The shifting balance theory and macroevolution. Annu Rev Genet 16: 1-19, 1982

    Google Scholar 

  11. Paulus W, Peiffer J: Intratumoral histologic heterogeneity of gliomas. A quantitative study. Cancer 64: 442-447, 1989

    Google Scholar 

  12. Coons SW, Johnson PC: Regional heterogeneity in the proliferative activity of human gliomas as measured by the Ki-67 labeling index. J Neuropathol Exp Neurol 52: 609-618, 1993

    Google Scholar 

  13. Shapiro JR, Ebrahim SAD, Mohamed AN, Pu PY, Shapiro WR: BCNU-sensitivity in parental cells and clones from four freshly resected near-diploid human gliomas: an astrocytoma, an anaplastic astrocytoma and two glioblastoma multiforme. J Neuro-Oncol 15: 209-227, 1993

    Google Scholar 

  14. Reifenberger G, Deckert M, Wechsler W: Immunohistochemical determination of protein kinase C expression and proliferative activity in human brain tumors. Acta Neuropathol 78: 166-175, 1989

    Google Scholar 

  15. Goldman S, Levivier M, Pirotte B, Brucher JM, Wikler D, Damhaut P, Stanus E, Brotchi J, Hildebrand J: Regional glucose metabolism and histopathology of gliomas. Cancer 78: 1098-1106, 1996

    Google Scholar 

  16. Shapiro JR, Yung WA, Shapiro WR: Isolation, karyotype, and clonal growth of heterogeneous subpopulations of human malignant gliomas. Cancer Res 41: 2349-2359, 1981

    Google Scholar 

  17. Coons SW, Johnson PC: Regional heterogeneity in theDNA content of human gliomas. Cancer 72: 3052-3060, 1993

    Google Scholar 

  18. Coons SW, Johnson PC, Shapiro JR: Cytogenetic and flow cytometry DNA analysis of regional heterogeneity in a low grade human glioma. Cancer Res 55: 1569-1577, 1995

    Google Scholar 

  19. Sidransky D, Mikkelsen T, Schwechheimer K, Rosenblum ML, Cavanee W, Vogelstein B: Clonal expansion of p53 mutant cells is associated with brain tumor progression. Nature 355: 846-847, 1992

    Google Scholar 

  20. Strommer K, Hamou MF, Diggelmann H, de Tribolet N: Cellular and tumoral heterogeneity of EGFR gene amplification in human malignant gliomas. Acta Neurochir (Wien) 107: 82-87, 1990

    Google Scholar 

  21. Chattopadhyay P, Rathore A, Mathur M, Sarkar C, Mahapatra AK, Sinha S: Loss of heterozygosity of a locus on 17p13.3, independent of p53, is associated with higher grades of astrocytic tumors. Oncogene 15: 871-874, 1997

    Google Scholar 

  22. Joshi AR, Sinha S, Dil-Afroze, Sulaiman IM, Banerji AK, Hasnain SE: Alterations in brain tumor DNA detected by a fingerprinting probe. Ind J Biochem Biophys 33: 455-457, 1996

    Google Scholar 

  23. Dil-Afroze, Misra A, Sulaiman IM, Sinha S, Sarkar C, Mahapatra AK, Hasnain SE: Alteration in brain tumor genomes identified by RAPD analysis. Gene 206: 45-48, 1998

    Google Scholar 

  24. Ong TM, Song B, Qian HW, Wu ZL, Whong WZ: Detection of genomic instability in lung cancer tissues by random amplified polymorphic DNA analysis. Carcinogenesis 19: 233-235, 1998

    Google Scholar 

  25. Wales MM, Biel MA, El Deiry WS, Nelkin BD, Issa JP, Cavanee WK, Kuerbitz SJ, Baylin SB: p53 activates expression of HIC-1, a new candidate tumor suppressor gene on 17p13.3. Nat Med 1: 570-577, 1995

    Google Scholar 

  26. Fidler IJ, Hart IR: Biological and experimental consequences of the zonal composition of solid tumours. Cancer Res 41: 3266-3267, 1981

    Google Scholar 

  27. Sambrook J, Fritsch EF, Maniatis T (eds): Molecular Cloning. Cold Spring Harbor Laboratory Press, New York, 1989

    Google Scholar 

  28. Largey JS, Meltzer SJ, Yin J, Norris K, Sauk JJ, Archibald DW: Loss of heterozygosity of p53 in oral cancers demonstrated by the polymerase chain reaction. Cancer 71: 1933-1937, 1993

    Google Scholar 

  29. Hahn M, Serth J, Fislage R, Wolfes H, Allhoff E, Jonas U, Pingoud A: Polymerase chain reaction detection of a highly polymorphic VNTR segment in intron 1 of human p53 gene. Clin Chem 39: 549-550, 1993

    Google Scholar 

  30. Carrozzo R, Ledbetter DH: Dinucleotide repeat polymorphism mapping to the critical region for lissencephaly (17p13.3). Hum Mol Genet 2: 615, 1993

    Google Scholar 

  31. Williams JGK, Hanafey MK, Rafalski JA, Tingey SV: Genetic analysis using random amplified polymorphicDNA markers. Method Enzymol 162: 704-740, 1993

    Google Scholar 

  32. Thein SL, Jeffreys AJ, Gooi HC, Cotter F, Flint J, O'Connor NTJ, Weatherall DJ, Wainscoat JS: Detection of somatic changes in human cancerDNAbyDNAfingerprinting analysis. Br J Cancer 55: 353-356, 1987

    Google Scholar 

  33. Matsumura Y, Tarin J: DNA fingerprinting survey of various human tumors and their metastases. Cancer Res 52: 2174-2179, 1992

    Google Scholar 

  34. Ellsworth DL, Rittenhouse KD, Honeycutt RL: Artifactual variation in randomly amplified polymorphic DNA banding patterns. Biotechniques 14: 214-217, 1993

    Google Scholar 

  35. Ayliffe MA, Lawrence GJ, Ellis JG, Pryor AJ: Heteroduplex molecules formed between allelic sequences cause nonparental RAPD bands. Nucleic Acid Res 22: 1632-1636, 1994

    Google Scholar 

  36. Micheli MR, Bova R, Calissano P, D'Ambrosio E: Randomly amplified polymorphic DNA fingerprinting using combinations oligonucleotide primers. Biotechniques 15: 388-390, 1993

    Google Scholar 

  37. Pillay M, Kenny ST: Anomalies in direct pair wise comparisons of RAPD fragments for genetic analysis. Biotechniques 19: 694-696, 698, 1995

    Google Scholar 

  38. Schweder ME, Shatteers RG, West SH, Smith RL: Effect of transition interval between melting and annealing temperature on RAPD analysis. Biotechniques 19: 38, 40-42, 1995

    Google Scholar 

  39. Arribas R, Capella G, Tortola S, Masramon L, Grizzle WE, Perucho M, Peinado MA:Assessment of genomic damage in colorectal cancer by DNA fingerprinting: prognostic applications. J Clin Oncol 15: 3230-3240, 1997

    Google Scholar 

  40. Caetano-Anolles G, Bassam BJ, Gresshoff PM: DNAampli-fication fingerprinting using very short arbitrary oligonucleotide primers. Biotechnology 9: 553-557, 1991

    Google Scholar 

  41. Peinado MA, Malkhosyan S, Velazquez A, Perucho M: Isolation and characterization of allelic losses and gains in colorectal tumors by arbitrarily primed polymerase chain reaction. Proc Natl Acad Sci USA 89: 10 065-10 069, 1992

    Google Scholar 

  42. Singh KP, Roy D: Detection of mutation(s) or polymorphic loci in the genome of experimental animal and human cancer tissues by RAPD/AP-PCR depends on DNA polymerase. Int J Oncol 14: 753-58, 1999

    Google Scholar 

  43. Daumas-Duport C: Patterns of tumor growth and problems associated with histological typing of low-grade gliomas. In: Benign Cerebral Glioma. Neurosurgical Topics. Apuzzo (ed) American Association of Neurological Surgeons, 1995, Vol 1, pp 125-147

    Google Scholar 

  44. Daumas-Duport C, Varlet P, Tucker ML, Beuvon F, Cervera P, Chodkiewicz JP: Oligodendrogliomas. Part I: Patterns of growth, histological diagnosis, clinical and imaging correlations: a study of 153 cases. J Neuro-Oncol 34: 37-59, 1997

    Google Scholar 

  45. Jung V, Romeike BFM, Henn W, Feiden W, Moringlane JR, Zang KD, Urbschat S: Evidence of focal genetic microheterogeneity in glioblastoma multiforme by area specific CGH on microdissected tumor cells. J Neuropath Exp Neurol 58: 993-999, 1999

    Google Scholar 

  46. Boehm T, Folkman J, Browder T, O'Reilly MS: Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 390: 404-407, 1997

    Google Scholar 

  47. Miller BE, Miller FR, Leith J, Heppner GH: Growth interaction in vivo between tumor subpopulations derived from a single mouse mammary tumor. Cancer Res 40: 3977-3981, 1980

    Google Scholar 

  48. Lantos PL, Vandenberg SR, Kleihues P: Tumors of the nervous system. In: Greenfield's Neuropathology. Graham DI, Lantos PL (eds) Arnold, London, 1997, Vol 2, pp 583-879

    Google Scholar 

  49. Shaw EG, Scheithauer BW, Gilbertson DT, Nichols DA, Laws ER, Earle JD, Daumas-Duport C, O'Fallon JR, Dinapoli RP: Postoperative radiotherapy of supra-tentorial low-grade gliomas. Int J Radiat Oncol Biol Phys 16: 663-668, 1989

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

    Anjan Misra & Parthaprasad Chattopadhyay

  2. Department of Pathology, All India Institute of Medical Sciences, New Delhi, India

    Amit K. Dinda & Subrata Sinha

  3. Department of Neurosurgery, All India Institute of Medical Sciences, New Delhi, India

    Chitra Sarkar & Ashok K. Mahapatra

  4. National Institute of Immunology, New Delhi, India

    Seyed E. Hasnain

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  1. Anjan Misra
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  2. Parthaprasad Chattopadhyay
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  3. Amit K. Dinda
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  6. Seyed E. Hasnain
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  7. Subrata Sinha
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Misra, A., Chattopadhyay, P., Dinda, A.K. et al. Extensive Intra-tumor Heterogeneity in Primary Human Glial Tumors as a Result of Locus Non-specific Genomic Alterations. J Neurooncol 48, 1–12 (2000). https://doi.org/10.1023/A:1006435201961

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