DNA methyltransferase 3B mutations linked to the ICF syndrome cause dysregulation of lymphogenesis genes

Abstract

ICF (immunodeficiency, centromeric region instability and facial anomalies) is a recessive disease caused by mutations in the DNA methyltransferase 3B gene (DNMT3B). Patients have immunodeficiency, chromosome 1 (Chr1) and Chr16 pericentromeric anomalies in mitogen-stimulated lymphocytes, a small decrease in overall genomic 5-methylcytosine levels and much hypomethylation of Chr1 and Chr16 juxtacentromeric heterochromatin. Microarray expression analysis was done on B-cell lymphoblastoid cell lines (LCLs) from ICF patients with diverse DNMT3B mutations and on control LCLs using oligonucleotide arrays for approximately 5600 different genes, 510 of which showed a lymphoid lineage-restricted expression pattern among several different lineages tested. A set of 32 genes had consistent and significant ICF-specific changes in RNA levels. Half of these genes play a role in immune function. ICF-specific increases in immunoglobulin (Ig) heavy constant mu and delta RNA and cell surface IgM and IgD and decreases in Ig(gamma) and Ig(alpha) RNA and surface IgG and IgA indicate inhibition of the later steps of lymphocyte maturation. ICF-specific increases were seen in RNA for RGS1, a B-cell specific inhibitor of G-protein signaling implicated in negative regulation of B-cell migration, and in RNA for the pro-apoptotic protein kinase C eta gene. ICF-associated decreases were observed in RNAs encoding proteins involved in activation, migration or survival of lymphoid cells, namely, transcription factor negative regulator ID3, the enhancer-binding MEF2C, the iron regulatory transferrin receptor, integrin beta7, the stress protein heme oxygenase and the lymphocyte-specific tumor necrosis factor receptor family members 7 and 17. No differences in promoter methylation were seen between ICF and normal LCLs for three ICF upregulated genes and one downregulated gene by a quantitative methylation assay [combined bisulfite restriction analysis (COBRA)]. Our data suggest that DNMT3B mutations in the ICF syndrome cause lymphogenesis-associated gene dysregulation by indirect effects on gene expression that interfere with normal lymphocyte signaling, maturation and migration.

A p38 MAPK-MEF2C pathway regulates B-cell proliferation

“…Dysregulation of B-cell proliferation can cause inadequate immune response, immunodeficiency, or leukemia (29,30). Our results highlight an important role for MEF2C in regulating B-cell proliferation.…”

DNA replication is altered in Immunodeficiency Centromeric instability Facial anomalies (ICF) cells carrying DNMT3B mutations

“…A similar variability was found in gene expression patterns, each patient having his own subset of dysregulated genes. [12][13][14][15] This variability reflects the heterogeneity of DNMT3B mutations and the resulting phenotypic and molecular variability that is observed among patients. Depending on the residual activity of the DNMT3B protein, each patient has his own transcriptome, and hence may have a different efficiency of DNA replication.…”

“…ICF-specific changes in RNA levels of genes critical for immune function, development, and neurogenesis were identified through the analysis of global expression profiles. [13][14][15][16] Finally, various late-replicating sequences (ie, satellite 2, telomeric repeats, and genes in F-heterochromatin) replicate earlier in ICF than in control cells. 9,11,17 In female ICF cells, genes located in F-heterochromatin lose DNA methylation and escape silencing, and this activation process is associated with a change in their replication timing from late to an earlier stage in the Sphase.…”

Epigenetics and Human Disease

“…3.3; Kondo et al 2000). Using lymphoblastoid cell lines of normal and ICF patients, gene expression studies revealed alterations in the expression of genes involved in maturation, migration, activation, and homing of lymphocytes (Ehrlich et al 2001). It is not clear from this study, however, whether loss of DNMT3B causes dysregulation of such genes because the methylation patterns at their promoter did not seem to be altered.…”