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Trial registered on ANZCTR


Registration number
ACTRN12617000500358
Ethics application status
Approved
Date submitted
7/02/2017
Date registered
6/04/2017
Date last updated
13/12/2017
Type of registration
Prospectively registered

Titles & IDs
Public title
NEST4E: A pilot study into the clinical effectiveness of Non-invasive Preimplantation Genetic Screening (PGS) method for Embryo ploidy status among patients undergoing IVF treatment
Scientific title
NEST4E: A pilot study into the clinical effectiveness of Non-invasive Preimplantation Genetic Screening (PGS) method for determining Embryo ploidy status among patients undergoing IVF treatment
Secondary ID [1] 291070 0
Nil
Universal Trial Number (UTN)
U1111-1192-4733
Trial acronym
NEST4E
Linked study record
none

Health condition
Health condition(s) or problem(s) studied:
Infertility 301868 0
Embryo genetics 301869 0
Condition category
Condition code
Reproductive Health and Childbirth 301546 301546 0 0
Fertility including in vitro fertilisation

Intervention/exposure
Study type
Interventional
Description of intervention(s) / exposure
The purpose of this pilot trial is to test a non-invasive Preimplantation Genetic Screening (PGS) method for human embryos. The investigators have developed a method for amplifying cell-free DNA from human embryo culture medium and using Next Generation Sequencing (NGS) to determine embryo aneuploid vs euploid status of the embryo, This technology eliminates the need for an invasive embryo biopsy to perform preimplantation genetic screening. This trial will be conducted at Repromed on 15 patients undergoing IVF treatment that would benefit from preimplantation genetic screening. Spent media will be collected by the scientists from the embryo culture dish and used to determine if the embryo is aneuploid or euploid. Each embryo undergoes this method once only on a single occasion, with the exception that if the non-invasive screen does not provide a conclusive result, a trophectoderm biopsy will be reformed on the embryo that fails the non-invasive screen to detect aneuploidy/euploidy.
Intervention code [1] 297059 0
Early detection / Screening
Comparator / control treatment
No control group
Control group
Uncontrolled

Outcomes
Primary outcome [1] 300946 0
The measured primary outcomes of this NEST4E pilot study are:
The number of embryos screened as euploid that are not required to be biopsied. This will be assessed by amplifying the cell-free DNA from the embryos spent culture media, The DNA will be sequenced using next generation sequencing and the data analysed using the Illumina platform to determine the ploidy status of the embryo. If the sequence data shows that the embryo is euploid the embryo will be transferred. The embryo will be confirmed as euploid at the primary time point (ie. following preimplantation and 1st trimester screening)
Timepoint [1] 300946 0
Following preimplantation and 1st trimester screening.
Primary outcome [2] 301479 0
For euploid embryos, the percentage of concordance of media sample results to non-invasive NIPT based first trimester screening which can be befriend as early as 10 weeks pregnant.
Timepoint [2] 301479 0
Following non-invasive NIPT based first trimester screening which can be performed as early as 10 weeks
Primary outcome [3] 301480 0
For anueploid embryos, the percentage of embryos that show concordance with confirmatory biopsy next generation sequencing screening results.
Timepoint [3] 301480 0
Next generation sequencing results of biopsied cells can be obtained with 2 weeks post trophectoderm biopsy
Secondary outcome [1] 331256 0
Measured secondary outcomes are:
Clinical pregnancy rates of embryos screened as euploid using the non-invasive NEST4E technology.
Timepoint [1] 331256 0
Determination of a positive pregnancy test occurs 12-14 days post embryo transfer via detection of HCG levels from a maternal blood test. Confirmation of a viable pregnancy is performed at 6-8 weeks of pregnancy via ulstrasound to detect fetal heart beat Live birth outcomes will be collected 2-4 weeks post the due date as per standard clinical protocol.,
Secondary outcome [2] 332790 0
Live birth rates of embryos screened as euploid using the non-invasive NEST4E test
Timepoint [2] 332790 0
Live birth outcomes will be collected as per standard Repromed patient care. This information is generally collected in writing from the treating obstetrician/doctor, or alternatively directly from the patient with a follow up phone call from the clinic nurses several weeks post due date. It is a regulatory requirement that gestation length, birthweight, sex, method of delivery and details of any maternal or neonatal complications are reported to ANZARD (Australian & New Zealand Assisted Reproduction Database).
Secondary outcome [3] 332791 0
Percentage of euploid vs aneuploidy embryos as determined by NEST4E non-invasive screening. Any embryos screened as Aneuploid will undergo trophectoderm biopsy and PGS screening to confirm the aneuploidy.
Timepoint [3] 332791 0
Data collection and analysis will be coordinated by the Principal Investigator and Co Investigators, all who have had extensive previous experience with data analysis and interpretation of pilot studies and clinical trials. These results will be reported directly to the Clinical Genetics Team and the patient’s clinician. A Data Monitoring Committee chaired by Repromed’s Clinical Director will perform an interim analysis after 5 participants embryo’s (approx 16-20 embryos) have been screened using NEST4E to ensure that the results are concordant with the biopsy screening. Final analysis will take place immediately after collection of the final birth outcome.

Eligibility
Key inclusion criteria
This study is a pilot study on fifteen couples (approximately 60 transferrable embryos) to assess the ability of this technology to screen embryos for aneuploidy. Participants will be consenting patients undergoing IVF treatment with PGS at Repromed, 180 Fullarton Rd., Dulwich, South Australia. Participation in the study will be offered to those patients whom PGS in general is discussed as a treatment option.
Minimum age
18 Years
Maximum age
53 Years
Gender
Both males and females
Can healthy volunteers participate?
Yes
Key exclusion criteria
1. Patients involved in the Donor program (oocyte and sperm)
2. Patients with a known infectious disease (HIV, HEP B, HEP C)
3. Patients using Preimplantation Genetic Diagnosis (PGD) for single genes or translocations.

Study design
Purpose of the study
Diagnosis
Allocation to intervention
Non-randomised trial
Procedure for enrolling a subject and allocating the treatment (allocation concealment procedures)
Allocation is not concealed
Methods used to generate the sequence in which subjects will be randomised (sequence generation)
non-randomised trial
Masking / blinding
Open (masking not used)
Who is / are masked / blinded?



Intervention assignment
Single group
Other design features
none
Phase
Not Applicable
Type of endpoint(s)
Efficacy
Statistical methods / analysis
This study is a pilot study on fifteen couples (approximately 60 transferrable embryos) to assess the ability of this technology to screen embryos for aneuploidy. Data collection and analysis will be coordinated by the Principal Investigator and Co Investigators. These results will be reported directly to the Clinical Genetics Team and the patient’s clinician.

Recruitment
Recruitment status
Active, not recruiting
Date of first participant enrolment
Anticipated
Actual
Date of last participant enrolment
Anticipated
Actual
Date of last data collection
Anticipated
Actual
Sample size
Target
Accrual to date
Final
Recruitment in Australia
Recruitment state(s)
SA
Recruitment hospital [1] 7435 0
Repromed Day Surgery - Dulwich
Recruitment postcode(s) [1] 15241 0
5065 - Dulwich

Funding & Sponsors
Funding source category [1] 295510 0
Commercial sector/Industry
Name [1] 295510 0
Repromed
Address [1] 295510 0
180 Fullarton Road Dulwich SA 5065
Country [1] 295510 0
Australia
Primary sponsor type
Commercial sector/Industry
Name
Repromed
Address
180 Fullarton Road Dulwich SA 5065
Country
Australia
Secondary sponsor category [1] 294329 0
None
Name [1] 294329 0
Address [1] 294329 0
Country [1] 294329 0

Ethics approval
Ethics application status
Approved
Ethics committee name [1] 296837 0
Bellberry Limited
Ethics committee address [1] 296837 0
129 Glen Osmond Road Eastwood South Australia 5063
Ethics committee country [1] 296837 0
Australia
Date submitted for ethics approval [1] 296837 0
27/01/2017
Approval date [1] 296837 0
09/03/2017
Ethics approval number [1] 296837 0
2016-12-891

Summary
Brief summary
The primary objective of this pilot study is to evaluate the clinical effectiveness of our NEST4E non-invasive method for screening embryo ploidy by determining the number of embryos that can that be successfully screened for aneuploidy using the NEST4E rather than invasive biopsy.
Aim 1: For euploid embryos the percentage concordance of media sample screening will be confirmed using non-invasive NIPT based first trimester screening as early as 10 week pregnancy. For aneuploid embryos, we will determine the percentage of embryos with concordance by confirmatory trophectoderm biopsy and PGS screening.
Aim 2: To determine clinical pregnancy rates and live birth outcomes of embryos that are transferred once screened as euploid.
Fifteen patients will be recruited by informed consent to participate in this pilot study to assess the clinical efficacy of our NEST4E non-invasive PGS test to screen their embryos for aneuploidy. Participants will be made aware that their participation in the study is entirely voluntary and that they can withdraw at any point in time. As per standard protocol, patient confidentially will be strictly maintained and their identity will not be revealed in any reviews and reports of this study which may be published.
Patients treatment will not be altered in any way and will proceed as per standard IVF cycle, including medications, oocyte retrieval, insemination and fertilization and embryo culture. Patient embryos will be cultured as per standard protocols; G1/G2 Plus media (Vitrolife) overlayed with oil (Ovoil, Vitrolife) in 6% CO2, 5% O2, 37 degrees. A 4 micro litre drop of the spent culture medium will be sampled using PCR grade filtered tips, under laminar flow, and placed into PCR grade 0.6 ml tubes and assessed for aneuploidy as described below.
DNA from the culture media sample will be amplified. If this amplification should fail, the embryo will be removed from the trial and will revert to standard care which involves a biopsy followed by vitrification and sample screening. If the amplification is successful, the embryo will be vitrified and the DNA amplified from the media sample used for screening by NEST4E. If conclusive sequence data is obtained, euploid embryos will be made available for transfer.
Trial website
Trial related presentations / publications
Public notes

Contacts
Principal investigator
Name 72182 0
Prof Michelle Lane
Address 72182 0
Repromed - 180 Fullarton Road Dulwich SA 5065
Country 72182 0
Australia
Phone 72182 0
+61 8 83338111
Fax 72182 0
Email 72182 0
mlane@repromed.com.au
Contact person for public queries
Name 72183 0
Dr Hamish Hamilton
Address 72183 0
Repromed - 180 Fullarton Road Dulwich SA 5065
Country 72183 0
Australia
Phone 72183 0
+61 8 83338111
Fax 72183 0
Email 72183 0
hhamilton@repromed.com.au
Contact person for scientific queries
Name 72184 0
Dr Hamish Hamilton
Address 72184 0
Repromed- 180 Fullarton Road Dulwich SA 5065
Country 72184 0
Australia
Phone 72184 0
+61 8 83338111
Fax 72184 0
Email 72184 0
hhamilton@repromed.com.au

No information has been provided regarding IPD availability
Summary results
Have study results been published in a peer-reviewed journal?
Other publications
Have study results been made publicly available in another format?
Results – basic reporting
Results – plain English summary