Development of the ERA-Microbiome Non-invasive Diagnosis Tool
| Status: | Active, not recruiting |
|---|---|
| Conditions: | Women's Studies |
| Therapuetic Areas: | Reproductive |
| Healthy: | No |
| Age Range: | 18 - 50 |
| Updated: | 3/27/2019 |
| Start Date: | August 4, 2017 |
| End Date: | February 2020 |
Development of a Non-invasive Diagnosis Tool for the Simultaneous Analysis of Endometrial Receptivity and Microbiota to Improve Reproductive Outcomes in Infertile Patients.
Although the endometrium tissue has been traditionally considered free of bacteria, recent
studies have documented the presence of an endometrial microbiome. In a pilot study conducted
by our research team, the microorganisms present in the endometrium were analysed in samples
of endometrial fluid (EF) using next generation sequencing (NGS).Consistent with previously
published studies, in normal conditions the endometrium is mainly composed of different
species of the genus Lactobacillus. It was further noted that the presence of other
pathogenic bacteria such as Streptococcus, and/or Gardnerella may alter the endometrial
microbiome and can disrupt the uterine environment, affecting implantation rates and
pregnancy success.
This project aims to validate the microbiome found in the endometrium of women of
reproductive age and try to corroborate the relationship between the endometrial microbiome
and the reproductive outcomes in patients undergoing assisted reproduction treatment (ART),
using the endometrial fluid samples to generate and validate a new diagnostic tool for
non-invasive endometrial receptivity. The use of Endometrial Receptivity Analysis (ERA) tool,
together with the analysis of the endometrial flora before the embryo transfer will allow to
evaluate the impact of the presence of endometrial pathogens on implantation in receptive
patients.
Therefore, the focus of this project is the development and clinical validation of a
non-invasive diagnosis tool to analyse simultaneously the endometrial receptivity and
microbiota, adding the microbiome study to the ERA analysis. This new tool will allow to
evaluate the endometrial factor in a comprehensive way by using non-invasive methods that
avoid endometrial biopsy (EB) collection and the discomfort caused to the patients.
studies have documented the presence of an endometrial microbiome. In a pilot study conducted
by our research team, the microorganisms present in the endometrium were analysed in samples
of endometrial fluid (EF) using next generation sequencing (NGS).Consistent with previously
published studies, in normal conditions the endometrium is mainly composed of different
species of the genus Lactobacillus. It was further noted that the presence of other
pathogenic bacteria such as Streptococcus, and/or Gardnerella may alter the endometrial
microbiome and can disrupt the uterine environment, affecting implantation rates and
pregnancy success.
This project aims to validate the microbiome found in the endometrium of women of
reproductive age and try to corroborate the relationship between the endometrial microbiome
and the reproductive outcomes in patients undergoing assisted reproduction treatment (ART),
using the endometrial fluid samples to generate and validate a new diagnostic tool for
non-invasive endometrial receptivity. The use of Endometrial Receptivity Analysis (ERA) tool,
together with the analysis of the endometrial flora before the embryo transfer will allow to
evaluate the impact of the presence of endometrial pathogens on implantation in receptive
patients.
Therefore, the focus of this project is the development and clinical validation of a
non-invasive diagnosis tool to analyse simultaneously the endometrial receptivity and
microbiota, adding the microbiome study to the ERA analysis. This new tool will allow to
evaluate the endometrial factor in a comprehensive way by using non-invasive methods that
avoid endometrial biopsy (EB) collection and the discomfort caused to the patients.
Embryonic implantation is a critical process to the survival of the species, that is
relatively inefficient especially in humans. Key elements are the embryo, the maternal
endometrium, and the cross-communication between them using different scientific
perspectives. Endometrial receptivity describes the phenotype that allows embryo adhesion and
placentation to occur. Due to the need of accurate, and objective molecular diagnostic test
for the receptivity status of endometrium the Endometrial Receptivity Analysis (ERA) was
developed. ERA test is a customized expression microarray that identifies the transcriptomic
expression profile signature of the personalized window of implantation (WOI). Its clinical
usefulness has been demonstrated in patients with implantation failure in whom 25% of them
have a displaced WOI becoming pregnant once the embryo transfer is performed at the proper
WOI predicted by ERA. However, this procedure requires an endometrial biopsy (EB), which is
the major drawback of this diagnosis test, because embryo transfer has to be delayed to the
next cycle, and also the possible discomforts caused to the patients. For this reason, a
previous pilot study was conducted in order to confirm whether endometrial receptivity can be
predicted through non-invasive methods in endometrial fluid (EF) samples. The gene panel of
ERA test was interrogated on epithelial and stromal cells from endometrial biopsies and cells
isolated from the EF at the single cell level, demonstrating that the major cells on the EF
classifies together with the epithelial population.
On the other hand, to try to better understand the mechanisms that connect endometrial
receptivity and/or implantation of the embryo with an altered microbiome endometrial, another
pilot study was conducted to determine the composition of the endometrial microbiome after
the analysis of the bacterial 16S ribosomal RNA by NGS. Interestingly, in patients with
receptive endometrium, diagnosed by Endometrial Receptivity Array (ERA) who had endometrial
microbiome with pathogens, or not dominated by bacteria of the genus Lactobacillus (NLD)
showed significantly lower implantation (23.1% vs 60.7% p = 0.02), pregnancy (33.3% vs 70.6%,
p = 0.03), ongoing pregnancy per embryo transfer (13.3% vs 58.8%, p = 0.03), and live birth
(6.7% vs 58.8%, p = 0.002) rates than those with a healthy endometrial microbiome dominated
by Lactobacillus (LD). Moreover, this relationship was much more significant when pathogenic
species found belonged to the genus Gardnerella and Streptococcus. This is in line with other
published studies that analyze the impact of endometrial pathogens in IVF treatments,
highlighting the importance of the study of bacterial communities for reproductive health.
Given these preliminary results, the present study aims to validate in a larger sample set,
the relationship between the imbalance of endometrial microbiome and the decline in
reproductive success in patients undergoing ART. To do this, the experimental design will be
improved in the following areas: (i) a higher sample size will be analysed to validate the
previous results obtained in the previous pilot study on endometrial microbiome, (ii) paired
samples from endometrial fluid and endometrial biopsy will be analysed for the microbial
profile to study whether there is bacterial variability associated to the sample type, (iii)
the endometrial microbiota will be simultaneously assessed with ERA as a supplementary
diagnosis for this tool, and (iv) the analysis of the endometrial microbiota will be
performed using the most advanced technology on bacterial metagenome sequencing to widen the
information of the microorganisms identified in each sample.
relatively inefficient especially in humans. Key elements are the embryo, the maternal
endometrium, and the cross-communication between them using different scientific
perspectives. Endometrial receptivity describes the phenotype that allows embryo adhesion and
placentation to occur. Due to the need of accurate, and objective molecular diagnostic test
for the receptivity status of endometrium the Endometrial Receptivity Analysis (ERA) was
developed. ERA test is a customized expression microarray that identifies the transcriptomic
expression profile signature of the personalized window of implantation (WOI). Its clinical
usefulness has been demonstrated in patients with implantation failure in whom 25% of them
have a displaced WOI becoming pregnant once the embryo transfer is performed at the proper
WOI predicted by ERA. However, this procedure requires an endometrial biopsy (EB), which is
the major drawback of this diagnosis test, because embryo transfer has to be delayed to the
next cycle, and also the possible discomforts caused to the patients. For this reason, a
previous pilot study was conducted in order to confirm whether endometrial receptivity can be
predicted through non-invasive methods in endometrial fluid (EF) samples. The gene panel of
ERA test was interrogated on epithelial and stromal cells from endometrial biopsies and cells
isolated from the EF at the single cell level, demonstrating that the major cells on the EF
classifies together with the epithelial population.
On the other hand, to try to better understand the mechanisms that connect endometrial
receptivity and/or implantation of the embryo with an altered microbiome endometrial, another
pilot study was conducted to determine the composition of the endometrial microbiome after
the analysis of the bacterial 16S ribosomal RNA by NGS. Interestingly, in patients with
receptive endometrium, diagnosed by Endometrial Receptivity Array (ERA) who had endometrial
microbiome with pathogens, or not dominated by bacteria of the genus Lactobacillus (NLD)
showed significantly lower implantation (23.1% vs 60.7% p = 0.02), pregnancy (33.3% vs 70.6%,
p = 0.03), ongoing pregnancy per embryo transfer (13.3% vs 58.8%, p = 0.03), and live birth
(6.7% vs 58.8%, p = 0.002) rates than those with a healthy endometrial microbiome dominated
by Lactobacillus (LD). Moreover, this relationship was much more significant when pathogenic
species found belonged to the genus Gardnerella and Streptococcus. This is in line with other
published studies that analyze the impact of endometrial pathogens in IVF treatments,
highlighting the importance of the study of bacterial communities for reproductive health.
Given these preliminary results, the present study aims to validate in a larger sample set,
the relationship between the imbalance of endometrial microbiome and the decline in
reproductive success in patients undergoing ART. To do this, the experimental design will be
improved in the following areas: (i) a higher sample size will be analysed to validate the
previous results obtained in the previous pilot study on endometrial microbiome, (ii) paired
samples from endometrial fluid and endometrial biopsy will be analysed for the microbial
profile to study whether there is bacterial variability associated to the sample type, (iii)
the endometrial microbiota will be simultaneously assessed with ERA as a supplementary
diagnosis for this tool, and (iv) the analysis of the endometrial microbiota will be
performed using the most advanced technology on bacterial metagenome sequencing to widen the
information of the microorganisms identified in each sample.
Inclusion Criteria:
- Patients in In Vitro Fertilization (IVF) / IntraCytoplasmic Sperm Injection (ICSI) or
OVODON with indication of ERA, with or without Preimplantation genetic screening (PGS)
indication, which receive embryo transfer with frozen blastocyst stage embryos (day
5/day 6) on a hormone replacement therapy (HRT) cycle.
- Maternal Age: ≤40 years (IVF/ICSI patients)
- Maternal Age: ≤50 years (OVODONATION patients)
- Body Mass Index (BMI): 18.5 - 30 kg / m2 (both inclusive)
- Negative serological tests for human immunodeficiency virus (HIV), hepatitis B virus
(HBV), hepatitis C virus (HCV), Rapid plasma reagin (RPR) for syphilis
- Women with regular menstrual formula (3-4 / 26- 35 days)
- Sperm concentration: > 2 million spermatozoa/ml
Exclusion Criteria:
- Patients who are Intra-Uterine Device (IUD) carriers in the last 3 months
- Patients who have taken prescribed antibiotics in the last 3 months before the sample
collection. (May be accepted those cases where the patient has received previous
prophylactic antibiotics to ovarian puncture in the stimulation cycle, according to
the standard clinical practice of the centre. The administration of this antibiotic
should be at least 1 month before the sample collection).
- Adnexal or uterine pathologies as uterine malformations that may affect the
implantation rate (e.g: Polyps, intramural myoma ≥ 4cm or submucosal, septum,
hydrosalpinx, etc). Patients affected (before or after of their inclusion) with any of
the previously described pathologies, will be allowed to participate in this study as
long as they are surgically treated before any study procedure.
- Existence of bacterial, fungal or viral infections severe or uncontrolled that, in the
opinion of the principal investigator, could interfere with the patient's
participation in the study or the assessments of the study results.
- Any illness or medical condition that is unstable or can put patient safety at risk
and compliance in the study.
We found this trial at
5
sites
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Chesterfield, Missouri 63005
Phone: 636-778-9900
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