A Study of Multilocus Methylation Abnormalities in Subjects With Imprinting Disorders

Imprinting disorders may be secondary to various molecular mechanisms, including "epimutations", i.e. aberrant methylation levels (gain or loss of methylation) at the level of the ICRs. The 11p15 region in humans contains two genes essential for controlling foetal growth: IGF2, expressed from the paternal allele, and whose expression is controlled by an imprinting centre called H19/IGF2: IG-DMR (or ICR1), methylated on the paternal allele; and CDKN1C, expressed from the maternal allele, and whose expression is controlled by a second imprinting centre called KCNQ1OT1:TSS-DMR (or ICR2), methylated on the maternal allele. (Eggermann et al, 2023) Among the diseases linked to parental imprinting, Beckwith Wiedemann syndrome (BWS, prevalence 1/13,500) is an overgrowth syndrome that, in particular, predisposes to the appearance of embryonic tumours during the first years of life. 80% of subjects with BWS have an alteration in the 11p15 region, which may be a loss of ICR2 methylation (around 50%) or a gain of ICR1 methylation (5 to 10%). (Brioude et al, 2018) Silver Russell syndrome (SRS, prevalence 1/50,000) is a syndrome of fetal and postnatal growth restriction, with preservation of head circumference, severe feeding difficulties and early metabolic complications. Approximately 50% of subjects with SRS have a loss of ICR1 methylation in the 11p15 region. (Wakeling et al, 2017). Temple syndrome (TS14) is a fetal and postnatal growth restriction syndrome similar to SRS with neonatal hypotonia, and associated with a risk of early puberty and obesity. This syndrome is usually caused by abnormalities of the 14q32 region at the MEG3/DLK1 locus which is also imprinted (maternal uniparental disomy, paternal deletion or loss of methylation of the MEG3/DLK1:IG-DMR). (Geoffron et al, 2017) For several years now, it has been shown that some subjects with a methylation anomaly at one locus may have methylation anomalies at other regions subject to parental imprinting. This condition is known as multilocus imprinting disorder (MLID). The prevalence of this multilocus disorder is not precisely known, because the methodologies used vary in terms of the technique used and the number of DMRs studied. In some subjects, symptoms of different diseases linked to these different regions may be present. However, little is currently known about the phenotypic consequences of these multilocus disorders. Finally, although most methylation anomalies occur sporadically, a few familial forms of methylation anomalies have been reported, with the common feature of recurrent miscarriages and the presence of multilocus disease in affected subjects. These familial forms have been associated with the presence in the mother of pathogenic variations in the genes encoding factors in the SCMC (subcortical maternal complex). Among other things, these factors are involved in oocyte maturation and the regulation of epigenetic marks after fertilisation and during the first cell divisions. (MacKay et al, 2024)

Inclusion criteria

• "Negative control" population: subjects who have undergone genetic exploration and:

• Having had a study of normal methylation of the 11p15 region (H19/IGF2: IG-DMR and KCNQ1OT1:TSS-DMR) by the reference technique.
• And having had a sequencing study of one or more genes involved in a growth pathology (Sanger sequencing and/or NGS analysis) and having concluded in the presence of a pathogenic variant responsible for the pathology.

OR

Study population:

• subjects who had undergone a reference genetic investigation (ASMM-RTqPCR) and for whom the final diagnosis was: gain or loss of methylation at the H19/IGF2:IG-DMR locus (BWS,SRS), at the KCNQ1OT1:TSS-DMR locus (BWS) or at the DLK1/MEG3:IG-DMR locus (TS14).
• Affiliation with a social security scheme or beneficiary (excluding AME).
• Signature of the consent form by the subject's parents or guardians, or the subject if of age at the time of the study.

Exclusion criteria

• Inability of either the subject (if over 18) or the parents or holder of parental authority to receive informed information about the protocol.
• Subject (if over 18) or parent or holder of parental authority deprived of their liberty by judicial or administrative decision