Cumulus Cell Mitochondrial Activity as a Non-invasive Marker of Embryo Quality (FLIM)

Cumulus Cell Mitochondrial Activity as a Non-invasive Marker of Embryo Quality

This study aims to test the extent of association of FLIM parameters of cumulus cells with clinical outcomes (clinical pregnancy, ongoing pregnancy at 12 weeks, and live birth) of the embryos that result from the enclosed oocytes.

Study Overview

• Status: Recruiting
• Conditions: Frozen Embryo Transfer; IVF/ICSI; Cumulus Cell; Fluorescence Lifetime Imaging Microscopy
• Intervention / Treatment: Genetic: FLIM parameters

Detailed Description

This study aims to test the extent of association of FLIM parameters of cumulus cells with clinical outcomes (clinical pregnancy, ongoing pregnancy at 12 weeks, and live birth) of the embryos that result from the enclosed oocytes.

Participants will be consented to and pre-signed following standard regimens of IVFMD, My Duc Hospital and IVFMD PN, My Duc Phu Nhuan Hospital. The standard is for patients with at least 8 follicles ≥ 10 mm on ultrasonography at the induced ovulation date.

Cumulus samples (CCs) will be collected following the protocol on the day of oocyte pick up (OPU), being assigned with the same number as the oocyte from which it was collected, and vitrified with a Cryotech kit until FLIM measurements are performed.

Insemination will be performed by using ICSI, 3 - 4 hours after oocyte retrieval. A sample of CCs will be sliced off the cumulus mass of each oocyte judged to be mature based on the cumulus mass disposition (absence of tight corona layers). The remaining cumulus cells will be stripped from the OCCs using hyaluronidase. Only mature oocytes will be inseminated.

The fertilization check will be performed under an inverted microscope for a period of 16-18 hours after insemination. On day 3, embryo evaluation will be performed at a fixed time point 66±2 hours after fertilization and, for day 5/6 transfers at 116±2 hours or 140±2 hours, respectively, using the Istanbul consensus. Freeze-all strategy for day 5 embryo will be applied and the first frozen embryo transfer cycle is single embryo transfer.

On the other hand, the investigators will stratify embryos into two groups based on the measured FLIM parameters from their associated cumulus cells, corresponding to the "high" 35% and "low" 65% of metabolic scores.

The investigators aim for embryos in the "high" metabolic scoring group to have a 10% higher live birth rate than controls. Assuming a current live birth rate of 32%, this corresponds to a hoped-for 42% live birth rate in the "high" group (and a 25% live birth rate in the "low" group). A total of 450 patients are required to detect an effect of this size (power 0.90, two-sided alpha 5%).

The primary endpoint of the study will be the live birth rate after the first embryo transfer of the started treatment cycle.

• Study Type: Observational
• Enrollment (Estimated): 200

Contacts and Locations

• Study Contact: Name: Tuong M HO, MD, MCE; Phone Number: +84903633377; Email: homanhtuong@yahoo.com
• Study Contact Backup: Name: Trang TM Ho, MSc; Phone Number: +84353242372; Email: trang.htm@myduchospital.vn

Study Locations

• Vietnam
  • Ho Chi Minh City
    • Ho Chi Minh, Ho Chi Minh City, Vietnam, 70000
      • Recruiting
      • My Duc Hospital
      • Contact: Minh Nguyen, MSc; Phone Number: +84973878491; Email: minh.nhn@myduchospital.vn

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No

• Sampling Method: Probability Sample

Study Population

All Vietnamese infertile women who have IVF/ICSI cycle with at least 8 follicles ≥ 10 mm on ultrasonography at induced ovulation date agreed to freeze-all day 5-6 embryos at IVFMD and IVFMD PN will be enrolled in the study.

Description

Inclusion Criteria:

• Having any number of IVF/ICSI attempts
• Ovarian stimulation with any stimulation protocol
• Patients with at least 8 follicles ≥ 10 mm on ultrasonography at induced ovulation date
• Agreed to freeze-all day 5-6 embryos
• Agreed to have a single day 5-6 embryo transfer.
• Agreed to participate in the research (signed the consent form)

Exclusion Criteria:

• In vitro maturation (IVM) cycles
• Cycles using testicular biopsy or epididymal sperm
• Cycles not having an oocyte freeze-all cycle
• Cycles using standard insemination
• Participating in another research

Study Plan

How is the study designed?

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort

Intervention / Treatment

Infertile women have IVF/ICSI cycle; All Vietnamese infertile women who have IVF/ICSI cycle with at least 8 follicles ≥ 10 mm on ultrasonography at induced ovulation date agreed to freeze-all day 5-6 embryos at IVFMD and IVFMD PN will be enrolled in the study.

Genetic: FLIM parameters; Cumulus samples (CCs) will be collected following the protocol on the day of oocyte pick up (OPU). CCs are frozen by the cryo-tech method, then stored in liquid nitrogen at -196oC. Information of CC samples and processed date/time is recorded. After all, samples are collected, they will be transferred to The Needleman Lab, Northwest Lab Building 359,52 Oxford Street Cambridge MA 02138 to measure mitochondria metabolism using FLIM.; Other Names: fluorescence lifetime imaging microscopy

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Live birth rate after one frozen embryo transfer cycle	The birth of at least one newborn, after 24 week's gestation that exhibits any sign of life, such as respiration, heartbeat, umbilical pulsation, or movement of voluntary muscles	After 24 week's gestation

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Ongoing pregnancy	Having at least 1 gestational sac on ultrasound at 12 weeks' gestation with heart beat activity	At 10 weeks after embryo placement
Positive pregnancy test	Positive pregnancy test is defined as a serum human chorionic gonadotropin level greater than 25 mIU/mL after2 weeks after embryo placement of the first transfer	At 2 weeks after embryo placement
Clinical pregnancy	The presence of at least one gestational sac on ultrasound at 7 week's gestation with the detection of heartbeat activity, after the completion of the first transfer	7 week's gestation
Implantation rate	Implantation rate is defined as the number of gestational sacs per number of embryos transferred 3 weeks after the first transfer	3 weeks after the first transfer

Collaborators and Investigators

• Sponsor: Mỹ Đức Hospital
• Collaborators: Harvard University Faculty of Medicine; The Needleman Lab, Northwest Lab Building 359,52 Oxford Street Cambridge MA 02138.; Hospital Foch, Suresnes, France.
• Investigators: Principal Investigator: Tuong M Ho, MD, MCE, Mỹ Đức Hospital

Publications and helpful links

General Publications

• Fauser BC. Towards the global coverage of a unified registry of IVF outcomes. Reprod Biomed Online. 2019 Feb;38(2):133-137. doi: 10.1016/j.rbmo.2018.12.001. Epub 2018 Dec 14. No abstract available.
• Dumollard R, Duchen M, Carroll J. The role of mitochondrial function in the oocyte and embryo. Curr Top Dev Biol. 2007;77:21-49. doi: 10.1016/S0070-2153(06)77002-8.
• May-Panloup P, Boucret L, Chao de la Barca JM, Desquiret-Dumas V, Ferre-L'Hotellier V, Moriniere C, Descamps P, Procaccio V, Reynier P. Ovarian ageing: the role of mitochondria in oocytes and follicles. Hum Reprod Update. 2016 Nov;22(6):725-743. doi: 10.1093/humupd/dmw028. Epub 2016 Aug 25.
• Babayev E, Seli E. Oocyte mitochondrial function and reproduction. Curr Opin Obstet Gynecol. 2015 Jun;27(3):175-81. doi: 10.1097/GCO.0000000000000164.
• Scott R 3rd, Zhang M, Seli E. Metabolism of the oocyte and the preimplantation embryo: implications for assisted reproduction. Curr Opin Obstet Gynecol. 2018 Jun;30(3):163-170. doi: 10.1097/GCO.0000000000000455.
• Chakraborty S, Nian FS, Tsai JW, Karmenyan A, Chiou A. Quantification of the Metabolic State in Cell-Model of Parkinson's Disease by Fluorescence Lifetime Imaging Microscopy. Sci Rep. 2016 Jan 13;6:19145. doi: 10.1038/srep19145.
• Becker W. Fluorescence lifetime imaging--techniques and applications. J Microsc. 2012 Aug;247(2):119-36. doi: 10.1111/j.1365-2818.2012.03618.x. Epub 2012 May 24.
• Sanchez T, Wang T, Pedro MV, Zhang M, Esencan E, Sakkas D, Needleman D, Seli E. Metabolic imaging with the use of fluorescence lifetime imaging microscopy (FLIM) accurately detects mitochondrial dysfunction in mouse oocytes. Fertil Steril. 2018 Dec;110(7):1387-1397. doi: 10.1016/j.fertnstert.2018.07.022. Epub 2018 Nov 14.
• Sanchez T, Zhang M, Needleman D, Seli E. Metabolic imaging via fluorescence lifetime imaging microscopy for egg and embryo assessment. Fertil Steril. 2019 Feb;111(2):212-218. doi: 10.1016/j.fertnstert.2018.12.014.

Helpful Links

• T. H. F. and E. A. (HFEA), "Fertility treatment 2019: trends and figures," 2021.

Study record dates

Study Major Dates

• Study Start (Actual): April 25, 2022
• Primary Completion (Estimated): June 30, 2024
• Study Completion (Estimated): March 30, 2025

Study Registration Dates

• First Submitted: April 10, 2022
• First Submitted That Met QC Criteria: April 10, 2022
• First Posted (Actual): April 18, 2022

Study Record Updates

• Last Update Posted (Actual): March 15, 2024
• Last Update Submitted That Met QC Criteria: March 14, 2024
• Last Verified: March 1, 2024

More Information

Terms related to this study

• Keywords: IVF/ICSI; FLIM; Cumulus cell; Day 5/6 frozen embryo; Fluorescence lifetime imaging microscopy
• Other Study ID Numbers: 11/21/DD/BVMD

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No