Clinical Trial Details
— Status: Not yet recruiting
Administrative data
| NCT number |
NCT05932082 |
| Other study ID # |
MIO study |
| Secondary ID |
|
| Status |
Not yet recruiting |
| Phase |
N/A
|
| First received |
|
| Last updated |
|
| Start date |
September 1, 2023 |
| Est. completion date |
September 1, 2026 |
Study information
| Verified date |
August 2023 |
| Source |
Peking Union Medical College Hospital |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Interventional
|
Clinical Trial Summary
In 2% to 3% infertility women, myoma is the only factor relevant to their infertility.
However, the effects of intramural myoma on fertility are controversial. For infertile women
with intramural myoma (types IV, V, VI of FIGO system) of 4 to 6 cm diameter, it is not clear
whether myomectomy could improve pregnancy outcomes, especially in women undergoing ART.
Besides, rigorous clinical research is needed to explore the changes and relevant biomarkers
of endometrial receptivity through multi-omics study in patients undergoing myomectomy and
ART treatment.
Method Intervention and follow-up: (1) For the control group, evaluation protocols such as
salpingography and/or laparoscopic tubal fluxation should be implemented to identify
disorders such as hydrosalpinx. (2) Imaging evaluation: all pelvic MRI were performed. Other
options such as transvaginal ultrasound are not excluded, but won't replace MRI. Enhanced MRI
or DWI may be considered, but are not always required. (3) Surgical intervention:
laparoscopic myomectomy is preferred, and abdominal myomectomy is also acceptable. (4) IVF
treatment: the IVF regimen should include detailed records of the downregulation plan, number
of cycles, frozen or fresh blastocysts.
Study endpoints
(1) Primary study endpoint: Live birth rate after IVF. (2) Secondary study endpoints:
Clinical pregnancy rate after IVF; Cumulative pregnancy rate after IVF; Biochemical pregnancy
rate after IVF; Sustained pregnancy rate after IVF (≥20 weeks); Miscarriage rate after IVF;
Cycles of IVF; Pregnancy-related complications; Perinatal maternal and neonatal
complications. (3) Exploratory endpoints: The correlation between imaging index and assisted
reproductive outcomes, including endometrial thickness, uterine volume, type of endometrial
echo, uterine contraction; endometrial vascular index (VI), flow index (FI), tubular flow
index (VFI); uterine artery pulsation index (PI), uterine artery resistance index (RI),
systolic/diastolic blood pressure of uterine artery (S/D).
The correlation between endometrial receptivity and assisted reproductive outcomes is
analyzed based on transcriptomics, metabolomics, methylation and proteomics in samples from
peripheral blood, endometrial biopsy, endometrial exfoliated cells, cervical exfoliated cells
and myoma.
Description:
Grouping and blinding method This is a national multicenter, randomized controlled study.
Competitive enrollment is performed among all centers. The study group received IVF after
myomectomy and the control group was directed to IVF after the routine evaluation, probably
including diagnostic laparoscopy. In this study, 792 patients were randomly divided into 2
groups with 396 patients in each group according to centralized random system. A random
sequence is generated with a random block size of a block size of 4.
Collection of clinical data and samples Data and samples are collected according to Case
Report Form (CRF) form, including randomized information, epidemiologic and clinical
information. Evaluation of infertility history and details, imaging results, ovarian
reservation and surgical findings are of special interest and detailed recorded.
Samples of peripheral blood, cervical exfoliated cells, endometrial exfoliated cells,
endometrial biopsy and myoma in study group are collected for multi-omics analysis. The
process of sample harvest shouldn't interfere or deteriorate the surgical and IVF treatment.
IVF treatment: the IVF regimen should include detailed records of the downregulation plan,
number of cycles, frozen or fresh blastocysts, ①Study group: IVF treatment is performed 6
months after myomectomy, and patients should be followed up for 1 year after the start of
IVF. If the endometrium is exposed during myomectoy, IVF treatment is performed 12 months
after myomectomy. It should be noted that the time for postoperative contraception should be
recorded, and also the time interval between myomectomy and IVF.
②Control group: IVF treatment was performed immediately after relevant evaluation, and
follow-up for 1 year after the start of IVF.
Genomic feature examination and bioinformatics analysis
1. Endometrial receptive array (ERA) is a molecular diagnostic test based on microarray
technology, which divides endometrial biopsy into receptive, pre receptive, or
proliferative types based on the expression of 238 selected genes. ER Map/ER Stage is
based on the expression of 184 genes involved in maternal immune response related to
endometrial proliferation and embryo implantation.
2. Methylation sequencing: Extract DNA related to peripheral blood, tissue, and cells, and
accurately quantify DNA concentration using Qubit 2.0. The total amount of DNA detected
should not be less than 1ug. Library construction: After passing the sample detection, 1
µ g of sample genomic DNA was mixed with unmethylated lambda DNA using the Biouptor
system, and then fragmented to an average size of approximately 250bp. After
fragmentation, the purified random fragmented DNA was then repaired, passivated and
phosphorylated with a mixture of T4 DNA polymerase, Klenow fragment and T4
polynucleotide kinase. The blunt DNA fragment was then 3 'adenosylated with Klenow
fragment (3' -5'exo -), and then linked to the linker connecting 5 '- methyl Cytosine
instead of Cytosine using T4 DNA ligase. After completing each step, use magnetic beads
to purify DNA. Then, use ZYMO EZ DNA methylation gold kit to convert unmethylated
Cytosine to Uracil according to instructions. Finally, use JumpStart Taq DNA polymerase
for PCR amplification, and then use magnetic beads to purify the PCR product to obtain
the final library. Library quality inspection: After the construction of the library is
completed, preliminary quantification is carried out using Qubit2.0, and the library is
diluted to 1ng/ul. Then, the insert size of the library is tested using Agilent 2100.
After the insert size meets expectations, the effective concentration of the library is
accurately quantified using qPCR method (effective concentration of the library>2nM) to
ensure library quality. Online sequencing: After passing the library detection,
different libraries are pooled according to the effective concentration and target
offline data volume requirements, and then sequenced on the Illumina Nova platform using
the PE150 sequencing strategy. After that, quality control of original offline data,
sequence comparison, calculation of methylation level, identification and statistics of
differential methylation region (DMR) were carried out.
Urinary protein detection and bioinformatics analysis
1. Protein separation technology: two-dimensional gel electrophoresis (2DE),
two-dimensional fluorescence differential electrophoresis (2D-DIGE), isoelectric
focusing electrophoresis (IEF), liquid chromatography; Multidimensional chromatography
(MDC) (including size exclusion chromatography, ion exchange chromatography, reverse
phase high performance chromatography, hydrophobic interaction chromatography, etc.),
and multidimensional liquid chromatography (MDLC).
2. Protein identification technology: primary mass spectrometry: according to different ion
sources, it can be divided into matrix assisted laser desorption ionization time of
flight mass spectrometry (MALDI-TOF-MS) and electric spray mass spectrometry (ESI MS);
Secondary mass spectrometry: peptide mass fingerprint (PMF); New technology: Stable
nuclide signature bio mass spectrometry (SIAMS).
3. Identification of protein peptide segments: amino and carboxyl end analysis, such as
Edman; Mass spectrometry, including peptide fragment mass spectrometry and tandem mass
spectrometry.
4. Protein interactions: Immunoprecipitation technology, Yeast two hybrid system, Protein
chip technology.
Metabolomics and Bioinformatics Analysis
1. The endometrium tissue was immediately frozen in liquid nitrogen after being isolated,
and then the tissue was stored at -80 ℃, ground and crushed, and extracted with cold
organic Liquid-liquid extraction. Attention must be paid to avoiding metabolic changes
before and during sampling. Perform metabolomics analysis on collected cells, tissues,
blood, and urine.
2. Metabolite extraction: The most commonly used method in metabolite extraction is to
precipitate protein with organic solvents, and extract the supernatant through
high-speed centrifugation or ultrafast filtration to remove protein precipitation.
Concentrate using freeze-drying or nitrogen blowing, and dissolve the residue in a
complex solvent.
3. Sample detection: Take a certain amount of redissolved supernatant and use Thermo
Vanquish UHPLC ultra-high performance liquid chromatography system and Thermo Q Active
HF-X mass spectrometry platform for machine detection.
4. Data processing: After the mass spectrometry detection is completed, the offline data (.
raw) file is imported into the search software for simple screening of retention time,
mass charge ratio, and other parameters. Then, according to the retention time
deviation, quality deviation and signal strength deviation, signal to noise ratio,
minimum signal strength and other information of different samples, peak alignment and
peak extraction are carried out. At the same time, the peak area is quantified, and then
the target ions are integrated to predict the Molecular formula and compare with the
database to obtain the identification and quantitative results of the data. Therefore,
non targeted metabolomics databases and search software play a decisive role in the
experimental results. However, LC-MS has very few public databases, and most of them are
platform built databases. The commonly used databases include HMDB, METLIN, and some
self built libraries.
5. Information analysis: Principal component analysis, differential metabolite analysis,
etc.
