Prolonged culture of normal chorionic villus cells yields ICF syndrome-like chromatin decondensation and rearrangements

Untreated cultures from normal chorionic villus (CV) or amniotic fluid-derived (AF) samples displayed dramatic cell passage-dependent increases in aberrations in the juxtacentromeric heterochromatin of chromosomes 1 or 16 (1qh or 16qh). They showed negligible levels of chromosomal aberrations in primary culture and no other consistent chromosomal abnormality at any passage. By passage 8 or 9, 82 ± 7% of the CV metaphases from all eight studied samples exhibited 1qh or 16qh decondensation and 25 ± 16% had rearrangements in these regions. All six analyzed late-passage AF cultures displayed this regional decondensation and recombination in 54 ± 16 and 3 ± 3% of the metaphases, respectively. Late-passage skin fibroblasts did not show these aberrations. The chromosomal anomalies resembled those diagnostic for the ICF syndrome (immunodeficiency, centromeric region instability, and facial anomalies). ICF patients have constitutive hypomethylation at satellite 2 DNA (Sat2) in 1qh and 16qh, generally as the result of mutations in the DNA methyltransferase gene DNMT3B. At early and late passages, CV DNA was hypomethylated and AF DNA was hypermethylated both globally and at Sat2. DNMT1, DNMT3A, or DNMT3B RNA levels did not differ significantly between CV and AF cultures or late and early passages. The high degree of methylation of Sat2 in late-passage AF cells indicates that hypomethylation of this repeat is not necessary for 1qh decondensation. Sat2 hypomethylation may nonetheless favor 1qh and 16qh anomalies because CV cultures, with their Sat2 hypomethylation, displayed 1qh and 16qh decondensation and rearrangements at significantly lower passage numbers than did AF cultures. Also, CV cultures had much higher ratios of ICF-like rearrangements to heterochromatin decondensation in chromosomes 1 and 16. These cultures may serve as models to help elucidate the biological consequences of cancer-associated satellite DNA hypomethylation.

[1]  T. Meitinger,et al.  DNA, FISH and complementation studies in ICF syndrome: DNA hypomethylation of repetitive and single copy loci and evidence for a trans acting factor , 1995, Human Genetics.

[2]  A. T. Sumner,et al.  ICF syndrome (immunodeficiency, centromeric instability and facial anomalies): investigation of heterochromatin abnormalities and review of clinical outcome , 1995, Human Genetics.

[3]  D. Smeets,et al.  ICF syndrome: a new case and review of the literature , 1994, Human Genetics.

[4]  C. Romano,et al.  Multibranched chromosomes 1, 9, and 16 in a patient with combined IgA and IgE deficiency , 1979, Human Genetics.

[5]  B. Dutrillaux,et al.  Segmentation of human chromosomes induced by 5-ACR (5-azacytidine) , 1976, Human Genetics.

[6]  C. E. Hildebrand,et al.  Human chromosome-specific repetitive DNA sequences: novel markers for genetic analysis , 2004, Chromosoma.

[7]  Christian B. Woods,et al.  Satellite DNA hypomethylation in karyotyped Wilms tumors. , 2003, Cancer genetics and cytogenetics.

[8]  M. Ehrlich,et al.  DNA methylation in cancer: too much, but also too little , 2002, Oncogene.

[9]  M. Ehrlich,et al.  High frequencies of ICF syndrome-like pericentromeric heterochromatin decondensation and breakage in chromosome 1 in a chorionic villus sample , 2001, Journal of medical genetics.

[10]  M. Fenech.  The role of folic acid and Vitamin B12 in genomic stability of human cells. , 2001, Mutation research.

[11]  T. de Ravel,et al.  The ICF syndrome: new case and update. , 2001, Genetic counseling.

[12]  C. Wijmenga,et al.  Genetic variation in ICF syndrome: Evidence for genetic heterogeneity , 2000, Human mutation.

[13]  M. Ehrlich,et al.  DNA hypomethylation and unusual chromosome instability in cell lines fromICF syndrome patients , 2000, Cytogenetic and Genome Research.

[14]  C. Wijmenga,et al.  The DNMT3B DNA methyltransferase gene is mutated in the ICF immunodeficiency syndrome. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Laird,et al.  CpG island hypermethylation in human colorectal tumors is not associated with DNA methyltransferase overexpression. , 1999, Cancer research.

[16]  M. Ehrlich,et al.  Frequent hypomethylation in Wilms tumors of pericentromeric DNA in chromosomes 1 and 16. , 1999, Cancer genetics and cytogenetics.

[17]  M. Ehrlich,et al.  Satellite DNA hypomethylation vs. overall genomic hypomethylation in ovarian epithelial tumors of different malignant potential. , 1999, Mutation research.

[18]  A. T. Sumner,et al.  A FISH study of chromosome fusion in the ICF syndrome: involvement of paracentric heterochromatin but not of the centromeres themselves. , 1998, Journal of medical genetics.

[19]  S. Baylin,et al.  Hypomethylation of pericentromeric DNA in breast adenocarcinomas , 1998, International journal of cancer.

[20]  M. Ehrlich,et al.  DNA demethylation and pericentromeric rearrangements of chromosome 1. , 1997, Mutation research.

[21]  M. Ehrlich,et al.  Preferential induction of chromosome 1 multibranched figures and whole-arm deletions in a human pro-B cell line treated with 5-azacytidine or 5-azadeoxycytidine. , 1997, Cytogenetics and cell genetics.

[22]  D. Eastmond,et al.  Detection of chromosomal breakage in the 1cen-1q12 region of interphase human lymphocytes using multicolor fluorescence in situ hybridization with tandem DNA probes. , 1995, Cancer research.

[23]  T. Haaf,et al.  The effects of 5-azacytidine and 5-azadeoxycytidine on chromosome structure and function: implications for methylation-associated cellular processes. , 1995, Pharmacology & therapeutics.

[24]  A. Niveleau,et al.  Abnormal methylation pattern in constitutive and facultative (X inactive chromosome) heterochromatin of ICF patients. , 1994, Human molecular genetics.

[25]  M. Jeanpierre.  Human satellites 2 and 3. , 1994, Annales de genetique.

[26]  F. Ledeist,et al.  An embryonic-like methylation pattern of classical satellite DNA is observed in ICF syndrome. , 1993, Human molecular genetics.

[27]  E. Viégas-Péquignot,et al.  Specific induction of uncoiling and recombination by azacytidine in classical satellite-containing constitutive heterochromatin. , 1993, Cytogenetics and cell genetics.

[28]  J. Bullerdiek,et al.  Chang medium raises the chromatin instability of pericentromeric areas of chromosome 1 in amniotic fluid cells , 1992, Prenatal diagnosis.

[29]  C. Fuster,et al.  Heterochromatin decondensation in chromosomes from chorionic villus samples , 1991, Prenatal diagnosis.

[30]  C. Fuster,et al.  Spontaneous chromosome fragility in chorionic villus cells. , 1991, Early human development.

[31]  F. Hecht,et al.  Chromosome abnormalities and genetic counseling. , 1990 .

[32]  F. Ledeist,et al.  Multibranched chromosomes in the ICF syndrome: immunodeficiency, centromeric instability, and facial anomalies. , 1989, American journal of medical genetics.

[33]  R. Blaese,et al.  Variable immunodeficiency with abnormal condensation of the heterochromatin of chromosomes 1, 9, and 16. , 1988, The Journal of pediatrics.

[34]  H. Willard,et al.  Chromosome-specific alpha satellite DNA from human chromosome 1: hierarchical structure and genomic organization of a polymorphic domain spanning several hundred kilobase pairs of centromeric DNA. , 1987, Genomics.

[35]  F. Harris,et al.  Centromeric instability of chromosomes 1 and 16 with variable immune deficiency: a new syndrome , 1985, Clinical genetics.

[36]  C. Gosden,et al.  Amniotic fluid cell types and culture. , 1983, British medical bulletin.

[37]  W. Chen.  Studies on the origin of human amniotic fluid cells by immunofluorescent staining of keratin filaments. , 1982, Journal of medical genetics.

[38]  M. Ehrlich,et al.  Amount and distribution of 5-methylcytosine in human DNA from different types of tissues of cells. , 1982, Nucleic acids research.

[39]  H. Hoehn,et al.  Morphological and biochemical heterogeneity of amniotic fluid cells in culture. , 1982, Methods in cell biology.