The primary objective of this study is to investigate if exogenously supplied ELABELA can induce MZ twinning in cultured human embryos. This will be done using time-lapse monitoring of donated human embryos that will be treated with synthetic…
ID
Source
Brief title
Condition
- Congenital and hereditary disorders NEC
Synonym
Research involving
Sponsors and support
Intervention
Outcome measures
Primary outcome
The main study parameter is the observation in at least 50% of the embryos
treated with exogenous ELABELA, followed by characterization, of postzygotic
splitting/duplication of the ICM compared to the control group (average MZ
twinning percentage ~2%).
Secondary outcome
NA
Background summary
Monozygotic (MZ) twinning is a very frequent phenomenon in humans with a
worldwide incidence of 1 every 300 births. Although not believed to be
genetically controlled, a few reports have documented familial MZ twinning. In
an extended Jordanian family with 16 pairs of monozygotic twins, the laboratory
of Bruno Reversade has identified a possible allele that predisposes to
monochorionic MZ twinning. Genetic analysis of these twins revealed chromosomal
linkage to a gene coding for a circulating hormone ELABELA (ELA). This hormone
was found to be overexpressed in iPSCs from these MZ twins and able to drive
the pluripotency and self-renewal of human Embryonic Stem Cells (hESCs) in
culture. We hypothesize that ELABELA added to a developing human blastocyst
will favor the survival of a second Inner Cell Mass (ICM), formed from single
ICM cells that remained attached to the opposite side of the blastocoel cavity
during expansion/collapse cycles or hatching, by preventing their demise
through inherent apoptosis.
Study objective
The primary objective of this study is to investigate if exogenously supplied
ELABELA can induce MZ twinning in cultured human embryos. This will be done
using time-lapse monitoring of donated human embryos that will be treated with
synthetic ELABELA relative to control embryos.
Should exogenous ELABELA suffice to induce morphological changes consistent
with monochorionic MZ twinning, the second objective of this study will be to
investigate what changes occur at the transcriptomic level upon ELABELA
treatment by performing single-cell RNA sequencing on developing
pre-implantation embryos. The observed transcriptomic changes will subsequently
be validated by qPCR-, RNAscope- and immunofluorescence- based assays.
Study design
Case-control study.
Study burden and risks
NA
de Boelelaan 1117
Amsterdam 1081 HV
NL
de Boelelaan 1117
Amsterdam 1081 HV
NL
Listed location countries
Inclusion criteria
Good quality embryos: According to standard laboratory protocol embryo transfer takes place on day 3 and surplus embryos are assessed again for developmental progression on day 4, day 5 and day 6. If the quality of the spare embryos is sufficient they are cryopreserved on day 4, day 5 or day 6. Embryos are considered for cryopreservation if the following criteria are met:
* Proof of fertilization: 2 pro-nuclei
* Regular development from day 1 to day 3
* Satisfactory embryo quality determined in terms of
o Number of blastomeres: morula on day 4, beginning of compaction on day 5 and blastocyst (B3) on day 6.
o Degree of fragmentation: maximal 20%
Only donated embryos of sufficient morphological quality after thawing will be considered for the study. The embryos have to fulfil the following criteria directly after thawing
o Minimum number of blastomeres: 8 cells
o Maximum degree of fragmentation: 20%
o Maximum degree of atresia: 25%
Exclusion criteria
A varying percentage of cell death (atresia) and developmental regression in thawed embryos is observed regularly. Therefore, embryos with insufficient morphological quality after thawing will be excluded from the study as described in the inclusion criteria.
Design
Recruitment
Followed up by the following (possibly more current) registration
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Other (possibly less up-to-date) registrations in this register
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In other registers
| Register | ID |
|---|---|
| CCMO | NL68209.000.18 |