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.