Clinical trial NCT00773825: Genomic Imprinting and Assisted Reproductive Technologies
Official title: Assessment of the Risk of Imprinting Defects in Children Born Following Assisted Reproductive Technologies (ART)
Study type: OBSERVATIONAL
Overall status: COMPLETED
Conditions: Natural Pregnancy
Pregnancy, Ovarian
Keywords: Genomic imprinting
Reproductive techniques, assisted
Beckwith-Wiedemann syndrome
Angelman syndrome
Lead sponsor: Assistance Publique - Hôpitaux de Paris
Sponsor class: OTHER
First posted: 2008-10-16
Enrollment: 542
Sex: FEMALE
Minimum age (years): 26
Maximum age (years): 40
Standard ages: ADULT
Accepts healthy volunteers: no
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: Women followed in a participating ART departments

Brief summary: Genomic imprinting, referring to an epigenetic marking resulting in monoallelic gene expression, plays a critical role in development. Recently, various imprinting diseases were reported in animals (Large Offspring syndrome (LOS)) and humans (Beckwith-Wiedemann syndrome (BWS) and Angelman syndrome (AS)) born after ART. In all cases, an imprinting defect was involved (loss of methylation at ICR2 in BWS, at SNRPN in AS and at IGF2R DMR2 in LOS). These data suggest that ART procedures may impair the establishment or the maintenance (following fertilization) of methylation marks at maternally imprinted loci. In view of these data, the aim of this study is to determine if children born following ART exhibit an increased risk of imprinting defects. If the answer is yes, the second objective is to identify the problematic step in the ART procedure and thus to suppress or modify this step.

Detailed description: Methodology: assessment of the methylation status at 9 different imprinted loci (using Southern blot and methyl-specific quantitative PCR) in 3 groups of patients: 150 children naturally conceived, 150 children conceived after ovarian stimulation but with in vivo fertilization, and 150 children conceived after ovarian stimulation and in VITRO fertilization. These analyses will be performed on cord blood. Fragments of placental tissue will also be collected for further analyses. Patients will be selected in maternity hospitals associated with ART departments ( ANTOINE BECLERE HOSPITAL, Cochin HOSPITAL, Saint-Vincent de Paul HOSPITAL, Jean VERDIER HOSPITAL, Tenon HOSPITAL and Dijon Hospital).

This work is also a unique opportunity to establish a DNA, RNA and tissue collection allowing further investigation regarding other epigenetic modifications than DNA methylation, not only at imprinted loci, but also in other genomic regions regulated by epigenetic modifications.

Eligibility criteria: Inclusion Criteria:

Mother :

* Age: 26 to 40 at conception
* Single foetus pregnancy
* Signed informed consent
* Affiliation to French health benefits
* Absence of maternal pathology
* Normal foetal karyotype (if available)
* Known procedure of ovarian stimulation
* ART procedure without sperm or oocyte donation
* ART in a participating ART departments
* Delivery in a participating hospital

Father

* Age : 18 to 50 at conception
* Signed informed consent

Exclusion Criteria:

* Abnormal foetal karyotype (if available)
* Delivery before 35 weeks of amenorrhea
* Delivery in not participating hospital
* Delivery complication leading to the absence of sample collection

Primary outcomes:
measure: Assessment of the methylation status at 9 imprinted loci in cord blood collected just after birth.
timeFrame: At the birth
Secondary outcomes:
measure: Assessment of other epigenetic marks (histone modifications) at imprinted loci and at non imprinted but epigenetically regulated loci.
timeFrame: At the birth
Other outcomes:
Locations:
city: Paris
country: France
facility: Trousseau Hospital
Study officials:
affiliation: Assistance Publique - Hôpitaux de Paris
name: Yves Le BOUC, PUPH
role: PRINCIPAL_INVESTIGATOR
References:
citation: Gicquel C, El-Osta A, Le Bouc Y. Epigenetic regulation and fetal programming. Best Pract Res Clin Endocrinol Metab. 2008 Feb;22(1):1-16. doi: 10.1016/j.beem.2007.07.009.
citation: Rossignol S, Steunou V, Chalas C, Kerjean A, Rigolet M, Viegas-Pequignot E, Jouannet P, Le Bouc Y, Gicquel C. The epigenetic imprinting defect of patients with Beckwith-Wiedemann syndrome born after assisted reproductive technology is not restricted to the 11p15 region. J Med Genet. 2006 Dec;43(12):902-7. doi: 10.1136/jmg.2006.042135. Epub 2006 Jul 6.
citation: Gicquel C, Gaston V, Mandelbaum J, Siffroi JP, Flahault A, Le Bouc Y. In vitro fertilization may increase the risk of Beckwith-Wiedemann syndrome related to the abnormal imprinting of the KCN1OT gene. Am J Hum Genet. 2003 May;72(5):1338-41. doi: 10.1086/374824. No abstract available.
citation: Gaston V, Le Bouc Y, Soupre V, Burglen L, Donadieu J, Oro H, Audry G, Vazquez MP, Gicquel C. Analysis of the methylation status of the KCNQ1OT and H19 genes in leukocyte DNA for the diagnosis and prognosis of Beckwith-Wiedemann syndrome. Eur J Hum Genet. 2001 Jun;9(6):409-18. doi: 10.1038/sj.ejhg.5200649.