Management of Abnormally Fertilized Zygotes? InVitro Correction of 3PN

Infertility Clinical Trial

Official title:

Treatment of Abnormally Multinucleated Zygotes Before Before Division Starting.

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT02358759
• Other study ID #: AlBarakaSD3
• Status: Completed
• Phase: Phase 0
• First received: January 15, 2015
• Last updated: January 11, 2016
• Start date: June 2014
• Est. completion date: September 2015

Study information

• Verified date: January 2016
• Source: Al Baraka Fertility Hospital
• Contact: n/a
• Is FDA regulated: No
• Health authority: Bahrain: Ethics CommitteeEgypt: Ministry of Health and Population
• Study type: Interventional

Clinical Trial Summary

In this newly developed protocol, and idea, is to manage those abnormally developed zygotes from different ART procedures.

The investigators developed the plan and requirements needed to select the target extra nucleus or pronuclei to be extruded from fertilized egg in order to maintain developing healthy normal embryo.

Description:

As approved by Dyban and Baranov et al, about 15-18% of abortions caused by triploidy fertilization. One of sources for maternally triploidy is failure in the first meiotic division (Jacobs et al., 1978).

One of Digynic triploidy is developed by fertilized giant oocyte (Dyban and Baranov, 1987) {nuclear but no cytoplasmic division in an oogonium or cytoplasmic fusion of two oogonia (Austin, 1960)}.

Giant oocyte characterized with bigger diameter and will distinguished polar bodies at metaphase II.

B. Rosenbusch et. al. 2002, cytogenetic study showed that extra haploid maternal copy associated with MII (46,XX/ 2N ) giant oocytes as well as triploidy with fertilized giant oocytes (3N with 69,XXX or 69,XXY).

First Mitotic division plane with polar axes studies by Scott, 2001 , shows that Pn developed closer to 2nd polar body is the maternal origin PN.

Giant oocytes were collected from different IVF cycles, to be injected with normal sperm using Intracytoplasmic sperm injection (ICSI). 18 hours post ICSI arranged for fertilization evaluation and PN removal for fertilized oocyte before syngamy starts.

Video attached shows process of zygote manipulation by the way avoiding the division axis and focusing the extra maternal PN to be aspirated.

Pronuclear transfer in human embryos for mitochondrial DNA correction started the methodology of pronuclear manipulation, for that possibility of utilizing of 3PNs developed embryos research tools can be started. We arranged to study available received giant oocytes during IVF cycles. Accordingly we arranged for pronuclear removal followed by FISH evaluation in order to targeting Normal males embryos that insure proper extra maternal pronucleus removal.

Successful trials of maternal PN removal for giant oocyte collected from different cases summarized in table 1. All blastocyst developed arranged for FISH, so all embryos were utilized for cytogenetic evaluation.

Recommendations:

Further evaluations using STRs (Short tandem repeat ) should be used for maternal-paternal genome differentiation. NGS study is under evaluation for developed embryos for full CCS reporting and more genetic integrity. Epigenetic evaluation study recommended for triploidy corrected embryos for genetic expressions and early embryo developments as well as differentiation between paternal and maternal genomic activity.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 22
• Est. completion date: September 2015
• Est. primary completion date: September 2015
• Accepts healthy volunteers: No
• Gender: Both
• Age group: 20 Years to 45 Years

Eligibility

Inclusion Criteria:

• Giant oocytes

• 3 PNs developed embryos at day 1 post ICSI.

Exclusion Criteria:

• Patient refused to involving their abnormal oocytes at our study.

Study Design

Allocation: Non-Randomized, Endpoint Classification: Safety Study, Intervention Model: Single Group Assignment, Masking: Single Blind (Outcomes Assessor), Primary Purpose: Treatment

Related Conditions & MeSH terms

• Infertility

Intervention

Procedure:
• Correction of abnormally fertilized oocytes
Removing of extra developed nucleus from fertilized oocyte.

Locations

Country	Name	City	State
Bahrain	Al Baraka Fertility Hospital	Manama	Adliyah
Egypt	Ibn Sina IVF Center- Ibn Sina Hospital	Sohag

Sponsors (1)

Lead Sponsor	Collaborator
Ahmad Mustafa Mohamed Metwalley

Countries where clinical trial is conducted

Bahrain,  Egypt, 

References & Publications (5)

Amato P, Tachibana M, Sparman M, Mitalipov S. Three-parent in vitro fertilization: gene replacement for the prevention of inherited mitochondrial diseases. Fertil Steril. 2014 Jan;101(1):31-5. doi: 10.1016/j.fertnstert.2013.11.030. Review. — View Citation

Jacobs PA, Angell RR, Buchanan IM, Hassold TJ, Matsuyama AM, Manuel B. The origin of human triploids. Ann Hum Genet. 1978 Jul;42(1):49-57. — View Citation

Rosenbusch B. The potential significance of binovular follicles and binucleate giant oocytes for the development of genetic abnormalities. J Genet. 2012;91(3):397-404. Review. — View Citation

Vogel G. Assisted reproduction. FDA considers trials of 'three-parent embryos'. Science. 2014 Feb 21;343(6173):827-8. doi: 10.1126/science.343.6173.827. — View Citation

Wolf DP, Mitalipov N, Mitalipov S. Mitochondrial replacement therapy in reproductive medicine. Trends Mol Med. 2015 Feb;21(2):68-76. doi: 10.1016/j.molmed.2014.12.001. Epub 2014 Dec 10. Review. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Other	Blastocyst development	availability of embryos to develop for day 5 giving chance for better cells evaluation and more genetic and cytogenetic studies.	Day 5 embryo development for blastocyst stage	Yes
Primary	Normal Embryos Developed	At 18 hours after ICSi nucleus to be removed in-order to make genetic correction for abnormally developed embryos. Final result is normal embryos produced with 2PNs evaluated by FSIH study for 5 chromosomes.; This indicated the genetic correction process was successful process, and applicable to produce normal growing embryos.	Day 1 or 18 hours post ICSI nucleus removal	Yes
Secondary	Day 3 embryo available for blastomer biopsy	Availability of successful nucleus removal from zygote to be developed to active dividing embryo available for cytogenetic study.; That indicated normal embryogenesis processing	1 Year	Yes
Secondary	5 chromosomes FISH study	chromosomes 13, 18, 21, X and Y to be screened using fluorescence insitu hybridization (FISH). In order to check primary euploid developed with male (XY) or female (XX) embryos.	1 Year	Yes