Ignite Creation Date:
2024-05-06 @ 8:21 PM
Last Modification Date:
2024-10-26 @ 3:25 PM
Study NCT ID:
NCT06346418
Status:
RECRUITING
Last Update Posted:
2024-04-04
First Post:
2024-03-20
Brief Title:
Maternal Genes and Epimutations Beckwith-Wiedemann Syndrome Reproductive Risks
Sponsor:
Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico
Organization:
Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico
Study Overview
Official Title:
Role of Maternal Effect Genes and Epimutations in Beckwith-Wiedemann Syndrome and Adverse Reproductive Outcomes
Status:
RECRUITING
Status Verified Date:
2024-03
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Pathogenic variants in subcortical maternal complex SCMC have been identified not only in mothers of Beckwith-Wiedemann syndrome BWS babies but also in women with reproductive disturbances such as failed pregnancy attempts and recurrent pregnancy loss Based on the higher incidence of BWS in children born from Assisted Reproductive Technology ART this project aims to investigate incidence and molecular mechanism of pathogenic variants of SCMC in women with reproductive disorders Study objectives will be i assess the incidence of these variants as a cause of differences in reproductive outcomes in the infertile female population and mothers of children with BWS ii identify methylation changes in women with reproductive problems including those with offspring affected by BWS iii determine the molecular causes underlying female infertility and imprinting disorder associated with damaging SCMC gene variants by employing a mouse model
Detailed Description:
The first aim of the project is to define the incidence of Maternal-Effect Genes MEGs and specifically of SCMC pathogenic variants as a cause of differences in reproductive outcomes in the infertile female population and mothers of BWS children To this aim three cohorts of patients will be recruited healthy women with offspring affected by BWS and peculiar reproductive history from our population of clinically and molecularly diagnosed BWS families Cohort 1 women under 35 undergoing ART for infertility defined as failure to achieve a pregnancy after 12 months or more of regular unprotected sexual intercourses and unable to obtain a live birth after three completed cycles or after the transfer of at least 6 blastocysts Cohort 2 and women under 35 with RPL Recurrent Pregnancy Loss defined as the loss of two or more pregnancies before 24 weeks of gestation Cohort 3
To identify pathogenic variants whole-exome sequencing WES will be performed as the first approach in all the recruited patients WES analysis will be carried out following various steps First the investigators will analyze different subsets of genes belonging to
1 Maternal effect genes as SCMC components and other related genes
2 Genes essential in the maturation of the oocyte and zygote progression through the early phases of the embryogenesis or highly expressed at different stages of oocyte maturation
3 Genes with known and potential roles in the establishment and control of genomic imprinting and involved in DNA methylation reactions
Subsequently variants with a high pathogenicity score will be analyzed to identify any genes that may be associated with the phenomenon but do not belong to the previously described categories of genes Finally the investigators will conduct a whole genome sequencing WGS analysis on a selected subgroup of BWS mothers with peculiar clinical histories and negative WES analysis to explore all the noncoding and regulatory regions not targeted by WES
The second aim of this project is to employ whole-genome methylation analysis to identify methylation changes in women with reproductive problems including those with offspring affected by BWS Specific tasks will be
1 Determining the whole-genome methylation of blood leukocytes of the cohorts of women described in Aim 1 and comparing it with that of a similar number of sex- and age-matched controls
2 Determining the whole-genome methylation of unfertilized oocytes derived from unsuccessful ART cycles of the same cohorts and comparing it with that of control oocytes derived from either donation for research or public datasets
DNA methylation will be determined in blood leukocytes by methylation array analysis and in unfertilized oocytes by single cell Bisulfite sequencing scBS-Seq
The third aim of the project is to determine the molecular mechanisms underlying female infertility and imprinting disorder associated with damaging SCMC gene variants by employing a mouse model Specific tasks will be
1 Determining the whole-genome methylation and RNA profiles of the Padi6 mutmut oocytes and pre-implantation embryos obtained after IVF with wild-type sperm
2 Transfer of the blastocysts derived from the IVF described in a into pseudopregnant females and analysis of whole genome DNA methylation and RNA in the derived mid-gestation embryos by BS-seq and RNA-seq
3 Mini-screening of epigenetics compounds on the 2-cell Padi6 mutmut embryos DNA methylation and RNA expression of the mouse oocytes and embryos will be determined by whole-genome scBS-seq and scRNA-seq
To identify pathogenic variants whole-exome sequencing WES will be performed as the first approach in all the recruited patients WES analysis will be carried out following various steps First the investigators will analyze different subsets of genes belonging to
1 Maternal effect genes as SCMC components and other related genes
2 Genes essential in the maturation of the oocyte and zygote progression through the early phases of the embryogenesis or highly expressed at different stages of oocyte maturation
3 Genes with known and potential roles in the establishment and control of genomic imprinting and involved in DNA methylation reactions
Subsequently variants with a high pathogenicity score will be analyzed to identify any genes that may be associated with the phenomenon but do not belong to the previously described categories of genes Finally the investigators will conduct a whole genome sequencing WGS analysis on a selected subgroup of BWS mothers with peculiar clinical histories and negative WES analysis to explore all the noncoding and regulatory regions not targeted by WES
The second aim of this project is to employ whole-genome methylation analysis to identify methylation changes in women with reproductive problems including those with offspring affected by BWS Specific tasks will be
1 Determining the whole-genome methylation of blood leukocytes of the cohorts of women described in Aim 1 and comparing it with that of a similar number of sex- and age-matched controls
2 Determining the whole-genome methylation of unfertilized oocytes derived from unsuccessful ART cycles of the same cohorts and comparing it with that of control oocytes derived from either donation for research or public datasets
DNA methylation will be determined in blood leukocytes by methylation array analysis and in unfertilized oocytes by single cell Bisulfite sequencing scBS-Seq
The third aim of the project is to determine the molecular mechanisms underlying female infertility and imprinting disorder associated with damaging SCMC gene variants by employing a mouse model Specific tasks will be
1 Determining the whole-genome methylation and RNA profiles of the Padi6 mutmut oocytes and pre-implantation embryos obtained after IVF with wild-type sperm
2 Transfer of the blastocysts derived from the IVF described in a into pseudopregnant females and analysis of whole genome DNA methylation and RNA in the derived mid-gestation embryos by BS-seq and RNA-seq
3 Mini-screening of epigenetics compounds on the 2-cell Padi6 mutmut embryos DNA methylation and RNA expression of the mouse oocytes and embryos will be determined by whole-genome scBS-seq and scRNA-seq
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None