Imprinting Alterations Research Articles

Frequent loss of imprinting of the H19 and IGF-II genes in ovarian tumors.

Several human imprinted genes have been identified and are implicated in genetic diseases and tumorigenesis. We studied alterations of two imprinted genes, the paternally imprinted H19 and maternally imprinted IGF2, in 15 ovarian tumors with various cell types. To know allele-specific expression of the two genes, we analyzed restriction fragment length polymorphisms (RFLPs) at the 3'-untranslated region (UTR) in their cDNA, compared with those in the respective genomic DNA. As a result, biallelic H19 and IGF2 expression was observed in 8 (62%) of 13 informative (heterozygous) ovarian cancers and in 6 of 11 informative cases, respectively. H19 loss of imprinting (LOI) was most frequently observed in malignant serous cystadenocarcinoma (in four of six cases), whereas IGF2 LOI was not common in malignant epithelial cancers because three of six such LOI events occurred in benign mucinous cystadenomas and non-cancerous endometriotic cyst. Our data suggest that the alteration of H19 and IGF2 imprinting plays differential roles in tumorigenesis and progression of ovarian tumors, depending on the tissue type as well as the developmental stage. Our data may argue against tumor suppressor activity of H19 in ovarian cancers.

Imprinting mutation in the Beckwith-Wiedemann syndrome leads to biallelic IGF2 expression through an H19-independent pathway.

The Beckwith-Wiedemann syndrome (BWS) is genetically linked to chromosome 11p15.5, and a variety of observations suggest that deregulation of imprinted genes in this region is causally involved in the pathogenesis of the disease. It has been shown that in some patients without cytogenetic abnormalities the otherwise repressed maternal copy of the insulin-like growth factor 2 (IGF2) gene is expressed, leading to biallelic expression of IGF2. In some of these cases, this is accompanied by repression and DNA methylation of the maternal (otherwise active) copy of the neighbouring H19 gene. Hence, it is attractive to think that mutations may interfere with some aspect of H19 imprinting, thus leading to an inactive maternal allele, and indirectly to activation of the maternal IGF2 allele as reported in mice with an H19 gene deletion. However, no mutations have been identified so far in these patients. The only known mutations associated with BWS are maternally transmitted translocations, which are clustered in two locations centrometric to IGF2. The first cluster is 200-400 kb from IGF2 and the second is several megabases away. Hence, genes located far from the translocation breakpoints are potentially deregulated by them. Here we provide the first evidence of alteration of imprinting in a translocation family, with biallelic expression of IGF2 and altered DNA replication patterns in the IGF2 region. Interestingly, H19 imprinting was normal, suggesting an H19-independent pathway to biallelic IGF2 transcription. DNA methylation in IGF2 remained monoallelic, suggesting that the mutation in this family had uncoupled allele-specific methylation from expression.

Distribution and changes in mu- and kappa-opiate receptors during the midlife neurodevelopmental period of Coho salmon, Oncorhynchus kisutch.

Parr-smolt transformation (PST) in coho salmon is associated with a plasma thyroid hormone (PT4) surge and a critical period of neural development that includes axonal sprouting, neurogenesis, and surges of select neurotransmitters. Here we provide a description of the selectivity, distribution, and the changes in the density of mu- and kappa-opiate receptors during PST, as revealed by quantitative in vitro autoradiography of [3H]Tyr-D-Ala-Gly-NMe-Phe-Gly-ol ([3H]DAMGO) and [3H]ethylketocyclazocine ([3H]EKC), respectively. The concentration of mu-receptors increased significantly in select cell groups in the early stages of parr-smolt transformation, until a peak was reached at the time coinciding with the peak of the PT4 surge. In other cell groups, the peaks occurred 1 or 2 weeks later. With one exception, this increase was followed by a decrease in concentration. The brain areas showing the highest concentrations are the dorsal nucleus of the ventral telencephalic area, the glomerular region, the granular layer of the valvula cerebelli, the nucleus diffuses of the inferior lobe, and the nucleus diffuses of the torus lateralis. Other regions with distinctly elevated mu-receptor concentrations are the stratum griseum centrale of the optic tectum and the preoptic area. The distribution of kappa-receptors is more diffuse, and the densities are considerably lower. The overlap in distribution of mu- and kappa-receptors is considerable, but significant exceptions are noted. For example, the dorsomedial nucleus of the dorsal telencephalic area, the habenular nucleus, and the dorsomedial nucleus of the thalamus exhibit a surge in density of kappa-receptors at the time of the PT4 surge, while the density of mu-receptors in these nuclei remain very low throughout parr-smolt transformation. The kappa-receptor containing cell groups are not identifiable until 3 weeks before the PT4 surge because of low densities. The most prominently labeled kappa-receptor regions are the ventral and dorsal nuclei of the ventral telencephalic area, the medial and dorsal zones of the dorsal telencephalic area, the optic tectum (all layers), the dorsomedial nucleus of the thalamus, the torus lateralis of the ventral hypothalamus, and the preoptic area. An increase of mu- and kappa-opiate receptor densities in specific brain regions may reflect roles in the alteration of brain organization, olfactory imprinting, neuroendocrine activity or other physiological activities. The overall distribution of these receptors are relatively more extensive in salmon than in other vertebrates so far studied.

Uniparental disomy and genomic imprinting as causes of human genetic disease.

The existence of parent-of-origin differences in the expression of some genes, a process known as genomic imprinting, has been recognized and documented over the past several years. This epigenetic marking process results in the differential expression of normal genes depending upon whether they were inherited from the mother or the father. A number of human disorders have been identified as resulting from alterations in genomic imprinting. One process which can unmask genomic imprinting is uniparental disomy, in which both members of a chromosome pair are contributed by one sex parent. When uniparental disomy is present, genetic abnormality can result either from homozygosity of a single mutant allele which is present in two doses, or from the presence of two copies of an imprinted unexpressed gene or genes, rather than the usual one expressed and one unexpressed. Examples of human genetic disorders that are the consequence of genomic imprinting, and a discussion of current knowledge about the mechanisms of imprinting and the causes of uniparental disomy, are reviewed.

Allelic methylation of H19 and IGF2 in the Beckwith — Wiedemann syndrome

Beckwith-Wiedemann syndrome (BWS) is a congenital overgrowth syndrome with associated embryonal tumours. Most BWS cases are sporadic but familial cases occur in 15% of patients and in these there is linkage to chromosome 11p15. In addition, a small number of patients have cytogenetic abnormalities involving chromosome 11p15. Approximately 20% of sporadic BWS patients have uniparental paternal disomy (UPD) of chromosome 11p15. This finding together with the observation that penetrance in familial cases depends on parental transmission, suggests that the gene(s) for BWS are imprinted. The recent demonstration of biallelic expression of the otherwise maternally imprinted IGF2 gene in some BWS patients implicates excess IGF2 expression in the disease. Here we have analysed the allele-specific methylation patterns in the IGF2 gene and in the neighbouring and reciprocally imprinted H19 gene in a group of 42 BWS patients, 10 of which were mosaic UPD cases. We found that allelic methylation of both genes was normal in all non-UPD cases, with the paternal allele being methylated, and was increased in UPD cases in proportion with the disomic lineage. These findings suggest that sporadic BWS is not associated with a general alteration of methylation imprinting of the IGF2 and H19 genes. The methylation assay used in this study thus also offers a simple and reliable diagnostic test of UPD for 11p15.5. An unexpected finding was a distortion of the frequency of AvaII alleles at the IGF2 locus exclusively in UPD BWS cases (P < 0.001). This further implicates the IGF2 gene in aspects of the BWS phenotype.