Multi-center Study to Validate niPGT-A

Brief Title

Multi-center Study to Validate niPGT-A

Official Title

A Prospective, Observational, Multi-center, International Study to Validate a Non-invasive Preimplantation Genetic Test for Embryo Aneuploidy in the Spent Culture Media (niPGT-A).

Brief Summary

      Abnormal chromosome number, or aneuploidy, is common in human embryos. It is responsible for
      more than half of all miscarriages, and it is the leading cause of congenital birth defects.
      Besides, it has been described that aneuploidy may also affect embryo implantation.
      Therefore, selecting embryos that have the best chance of implanting and growing into a
      healthy baby is one of the most important steps in the field of assisted reproduction.

      Recent advances in genetic technologies, such as Next-Generation Sequencing (NGS), have
      allowed aneuploidy to be detected with greater sensitivity. The application of this technique
      to trophectoderm biopsies, taken from embryos before transfer to the uterus, has provided
      insight into the clinical impact of chromosomal status. This process of screening embryos to
      make sure they have the right number of chromosomes and to look for any structural
      abnormalities in the chromosomes is called Preimplantation Genetic Testing for Aneuploidy
      (PGT-A). It requires specific equipment and trained personnel that will add costs and risks,
      so non-invasive techniques are sought as an alternative. These non-invasive procedures has
      been explored by some groups analyzing the spent culture medium where the embryo is incubated
      up to the time of transfer or freezing. In daily routine, this media is discarded after
      finishing the embryo culture, but it has been reported that contains traces of embryonic
      cell-free DNA (cfDNA) that can represent the genetic load of the embryo. However, at the
      moment there is a high variability in results across studies, with a percentage of concordant
      results between the media and the trophectoderm biopsy ranging from 3.5 to 85.7%.

      Thus, the main objective of this project is to validate a new non-invasive method for PGT-A
      (niPGT-A), based on improved collection and analysis of the culture media to achieve higher
      rates of sensitivity and specificity and to decrease the effect of some intrinsic
      difficulties such as low embryonic cfDNA input, mosaicism and maternal contamination.
    

Detailed Description

      Human embryos have higher aneuploidy rates (20-80%) than other species. A considerable
      proportion of these aneuploid embryos have the ability to reach the blastocyst stage.
      However, depending on the aneuploidy type, some will fail to implant in the uterus, while
      others will implant but will be unable to carry out early embryonic development
      (miscarriage), or very rarely, result in liveborn children with specific abnormalities. It is
      therefore important to identify aneuploid embryos It is important to identify embryos at risk
      for aneuploid chromosomes in patients with advanced maternal age (AMA), recurrent
      implantation failure (RIF) or recurrent miscarriage (RM).

      The identification of aneuploidies is especially important in embryos from patients with
      higher aneuploidy risk such as those with advanced maternal age (AMA), recurrent implantation
      failure (RIF), or recurrent miscarriage (RM). Therefore, normal embryo morphology and
      development are dependent on the chromosomal complement.

      PGT-A technique analyse the full chromosome content of a single cell with high sensitivity
      and specificity but requires an invasive biopsy to obtain embryonic material for the genetic
      analysis. Thus, non-invasive methods to replace the existing invasive testing method would be
      useful in the improvement of maternal and fetal safety.

      Recently, there have been many research advances in the field of genetic testing. Cell-free
      DNA (cfDNA) has been observed in spent embryo culture media. The origin of the cfDNA at the
      blastocyst stage could be attributed to apoptotic events which may occur during normal
      development. This has encouraged different research groups to carry out analysis spent
      culture media.

      Various studies were initially carried out to detect specific genes associated with monogenic
      disorders (MTHFR9, HBA1/HBA210, SRY11). Recently, non-invasive PGT-A has been developed, with
      highly variable results on the concordance rate (3.5%,59.1%, and 85.7%, 30.6%). The
      chromosomal status of the embryo from the DNA present in the spent culture medium was
      compared to the one obtained following the standard protocol using trophectoderm biopsy. The
      difference in the reported results can be related to the different methodologies applied
      because different amplification and detection methods -aCGH or NGS- were used. Moreover, the
      concordance rates were defined differently on each study, i.e. aneuploid results in spent
      culture media and trophectoderm biopsy could be considered concordant despite of showing not
      the same aneuploid chromosomes.

      The impact of culture conditions in the efficiency of the non-invasive approach has been
      investigated by Hammond et al, (2016). They found that there was consistently a very low
      level of DNA contamination (mitochondrial and nuclear DNA) in media controls, from three
      different types of commercial media, that had not been exposed to embryos. The low baseline
      level of DNA contamination observed is thought to originate from the protein supplement of
      the culture media.

      Finally, there could be influence of two relevant factors on PGT-A: contamination with
      maternal DNA from granulosa cells (MCC) and mosaicism.

      To improve the results of IVF (In vitro Fertilization) programs, there is a need to identify
      the embryo with highest implantation potential. Embryo chromosomal analysis allows the
      selection of euploid embryos, which have a higher implantation success rate.

      The development of a non-invasive PGT-A protocol will improve the current methodologies used
      to identify those euploid embryos avoiding the detrimental effect of the biopsy on the embryo
      and decreasing the economic cost.

      The initial estimated sample size calculated was 3245 embryos (each embryo is considered as a
      subject in the study), considering a dropout rate of 30%.

      Data will be grouped and analyzed at Igenomix at three time points of the study: once 25
      embryos of each center have been processed (to assess the implementation of the methodology),
      after the 30% of the samples have been processed (as an interim to check the results) and at
      the end of the study for the final analysis including the follow up of the clinical outcome.

      After the interim analysis performed at 30% of recruitment, the total sample size has been
      recalculated as 2620 samples, considering a drop-out rate of 5% according to the drop-out
      rate observed during the interim analysis. Results of the interim analysis published in Rubio
      et al., AJOG 2020.
    


Study Type

Observational


Primary Outcome

Chromosomal status of the embryos

Secondary Outcome

 Pregnancy rate

Condition

Aneuploidy

Intervention

PGT-A

Study Arms / Comparison Groups

 Embryos undergoing PGT-A / niPGT-A
Description:  Embryos from IVF patients between 20 and 44 years of age, undergoing PGT-A for any medical indication, with own oocytes or ovum donation cycles and with single embryo transfer (SET)

Publications

* Includes publications given by the data provider as well as publications identified by National Clinical Trials Identifier (NCT ID) in Medline.

Recruitment Information


Recruitment Status

Diagnostic Test

Estimated Enrollment

2620

Start Date

April 27, 2018

Completion Date

September 2021

Primary Completion Date

September 2021

Eligibility Criteria

        Inclusion Criteria:

          -  PGT-A cases with trophectoderm biopsy and SET for any medical indication and signed
             written informed consent form approved by the EC/IRB after having been duly informed
             of the nature of the research and voluntarily accepted to participate in the study.

          -  ICSI (Intra Cytoplasmic Sperm Injection), IVF (In Vitro Fertilization) or ICSI/IVF
             performed in fresh oocytes from couples are allowed.

        Note: Donor sperm is allowed.

          -  Only fresh oocytes allowed.

          -  Fresh and Deferred Embryo Transfer are allowed. Note: In case of Deferred Embryo
             Transfer, embryos must be vitrified always after the blastocyst biopsy.

          -  Age: 20-44 years of age (both included).

        Exclusion Criteria:

          -  A known abnormal karyotype in a member of the couple.

          -  Preimplantation Genetic Testing for Monogenic diseases (PGT-M) or Preimplantation
             Genetic Testing for Structural Rearrangements (PGT-SR) cases excluded.
      

Gender

Female

Ages

20 Years - 44 Years

Accepts Healthy Volunteers

No

Contacts

Carmen Rubio, BSc PhD, +34963905310, [email protected]

Location Countries

Argentina

Location Countries

Argentina

Administrative Informations


NCT ID

NCT03520933

Organization ID

IGX1-NIP-CS-18-02-SUB1


Responsible Party

Sponsor

Study Sponsor

Igenomix


Study Sponsor

Carmen Rubio, BSc PhD, Principal Investigator, Igenomix


Verification Date

January 2021