Human germline nuclear transfer to overcome mitochondrial disease and failed fertilization after ICSI

Tang, Manshu; Boel, Annekatrien; Castelluccio, Noemi; Barberán, A Cardona; Christodoulaki, A; Bekaert, Bieke; Popovic, Mina; Meerschaut, Frauke Vanden; Sutter, Petra De; Menten, Björn; Symoens, Sofie; Vanlander, Arnaud; Stoop, Dominic; Coucke, Paul; Heindryckx, Björn

doi:10.1007/s10815-022-02401-7

Cited by 14 publications

(7 citation statements)

References 55 publications

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“…It was previously reported that these oocytes exhibit low fertilization and embryo potential ( Lierman et al , 2021 ), probably owing to cytoplasmic factors, which nevertheless was partially overcome by the application of ST ( Christodoulaki et al , 2023 ). The technology of NT has also been considered for certain infertility indications, including failed fertilization and embryo developmental arrest in both animal and human studies ( Zhang et al , 2016 ; Costa-Borges et al , 2020 ; Tang et al , 2020 , 2022 ). Until recently, the only live birth in human from NT was reported by Zhang et al (2017) in a case study involving a patient with a mitochondrial DNA mutation, leading to a healthy boy.…”

Section: Discussionmentioning

confidence: 99%

“…Several NT techniques have been developed, but the most studied ones are spindle transfer (ST) and pronuclear transfer (PNT) ( Christodoulaki et al , 2021 ). To date, one healthy baby has been born after the application of ST to overcome mitochondrial DNA diseases ( Zhang et al , 2017 ), while lately it has also been considered as a means to overcome certain infertility indications, such as fertilization failure and embryo developmental arrest in mice and human ( Costa-Borges et al , 2020 ; Tang et al , 2020 , 2022 ; Costa-Borges et al , 2023 ). Finally, the use of NT for female-related infertility recently led to the first live births in human, at a very high efficiency ( Costa-Borges et al , 2023 ).…”

Section: Introductionmentioning

confidence: 99%

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Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles

Christodoulaki,

He,

Zhou

et al. 2024

Human Reproduction Open

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STUDY QUESTION Is pronuclear transfer (PNT) capable of restoring embryo developmental arrest caused by cytoplasmic inferiority of in vitro grown (IVG) mouse oocytes? SUMMARY ANSWER PNT to in vivo matured cytoplasm significantly improved embryo development of IVG mouse oocytes, leading to living, fertile offspring. WHAT IS KNOWN ALREADY In vitro follicle culture has been considered as a fertility preservation option for cancer patients. Studies describing the culture of human follicles remain scarce, owing to low availability of tissue. Mouse models have extensively been used to study and optimize follicle culture. Although important achievements have been accomplished, including the production of healthy offspring in mice, IVG oocytes are of inferior quality when compared to in vivo grown oocytes, likely because of cytoplasmic incompetence. STUDY DESIGN, SIZE, DURATION The study was carried out form September 2020 to February 2022. In total, 120 15-day-old B6D2 mice were used to perform secondary follicle culture and assess the quality of IVG oocytes. In vivo grown control oocytes were obtained from 85 8-12-week-old B6D2 mice, following ovarian stimulation. For sperm collection, four B6D2 males between 10 and 14 weeks old were used. For embryo transfer, 14 8-12 week-old CD1 females served as surrogate mothers and 10 CD1 vasectomized males 10-24 weeks old were used to generate pseudo-pregnant females. Finally, for mating, four B6D2 female mice aged 8-10 weeks and two B6D2 male mice 10 weeks old were used to confirm the fertility of nuclear transfer-derived pups. PARTICIPANTS/MATERIALS, SETTING, METHODS Secondary follicles from 15-day-old B6D2 mice were isolated from the ovaries and cultured for 9 days, before a maturation stimulus was given. Following 16-18 h of maturation, oocytes were collected and evaluated on maturation rate, oocyte diameter, activation rate, spindle morphology, calcium releasing ability and mitochondrial membrane potential. For every experiment, in vivo grown oocytes were used as a control for comparison. When cytoplasmic immaturity and poor embryo development were confirmed in IVG oocytes, PNT was performed. For this, the pronuclei from IVG oocytes, created following parthenogenetic activation and IVF, were transferred to the cytoplasm of fertilized, in vivo grown oocytes. Genetic analysis and embryo transfer of the generated embryos were implemented to confirm the safety of the technique. MAIN RESULTS AND THE ROLE OF CHANCE Following 9 days of follicle culture, 703 oocytes were collected, of which 76% showed maturation to metaphase II stage. Oocyte diameters were significantly lower in IVG oocytes, measuring 67.4 μm versus 73.1 μm in controls (p < 0.001). Spindle morphology did not differ significantly between IVG and control oocytes, but calcium-releasing ability was compromised in the IVG group. An average calcium release of 1.62 arbitrary units (AU) was observed in IVG oocytes, significantly lower than 5.74 in control oocytes (p < 0.001). Finally, mitochondrial membrane potential was inferior in IVG compared to the control group, reaching an average value of 0.95 versus 2.27 (p < 0.001). Developmental potential of IVG oocytes was assessed following parthenogenetic activation with strontium chloride (SrCl2). Only 59.4% of IVG oocytes cleaved to two cells and 36.3% reached blastocyst stage, significantly lower than 89.5% and 88.2% in control oocytes, respectively (p < 0.001 and p < 0.001). Both PNT and spindle transfer (ST) were explored in pilot experiments with parthenogenetically activated oocytes, as a means to overcome poor embryo development. After the added value of nuclear transfer (NT) was confirmed, we continued with the generation of biparental embryos by PNT. For this purpose, IVG and control oocytes first underwent IVF. Only 15.5% of IVG oocytes were normally fertilized, in contrast to 45.5% in controls (p < 0.001), with resulting failure of blastocyst formation in the IVG group (0% versus 86.2%, p < 0.001). When the pronuclei of IVG zygotes were transferred to the cytoplasm of control zygotes, the blastocyst rate was restored to 86.9%, a similar level as the control. Genetic analysis of PNT embryos revealed a normal chromosomal profile, to a rate of 80%. Finally, the generation of living, fertile offspring from PNT was possible following embryo transfer to surrogate mothers. LARGE SCALE DATA N/A LIMITATIONS, REASONS FOR CAUTION Genetic profiles of analysed embryos from PNT originate from groups that are too small to draw concrete conclusions, whilst ST, which would be the preferred NT approach, could not be used for the generation of biparental embryos owing to technical limitations. Even though promising, the use of PNT should be considered as experimental. Furthermore, results were acquired in a mouse model, so validation of the technique in human IVG oocytes needs to be performed to evaluate the clinical relevance of the technology. The genetic profiles from IVG oocytes, which would be the ultimate characterization for chromosomal abnormalities, were not analysed owing to limitations in the reliable analysis of single cells. WIDER IMPLICATIONS OF THE FINDINGS PNT has the ability to overcome the poor cytoplasmic quality of IVG mouse oocytes. Considering the low maturation efficiency of human IVG oocytes and potential detrimental effects following long-term in vitro culture, NTcould be applied to rescue embryo development and lead to an increased availability of good quality embryos for transfer. STUDY FUNDING/COMPETING INTEREST(S) A.C. is a holder of FWO (Fonds voor Wetenschappelijk Onderzoek) grants (1S80220N and 1S80222N). B.H. and A.V.S. have been awarded with a special BOF (Bijzonder Onderzoeksfonds), GOA (Geconcerteerde onderzoeksacties) 2018000504 (GOA030-18 BOF) funding. B.H. has been receiving unrestricted educational funding from Ferring Pharmaceuticals (Aalst, Belgium). The authors declare that they have no conflict of interest.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles

Christodoulaki,

He,

Zhou

et al. 2024

Human Reproduction Open

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“…In support of the relevance of a mature optimal ooplasm, a recent study used meiotic spindle transplantation into donor ooplast to overcome failed fertilization while still allowing women to have genetically-related offspring ( 22 ). This technique offered an alternative usage of donor oocytes, not only for their mitochondria but solely for their competent cytoplasm to correct fertilization failure while allowing patients to have their own progenies.…”

Section: Discussionmentioning

confidence: 99%

Assisted gamete treatment to pinpoint acquired meiotic maturity and overcome oocyte activation deficiency contributed by both gametes

Xie¹,

Aluko²,

Cheung³

et al. 2023

F&S Reports

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“…Although MII‐NT emerged as MR therapy, the possibility of providing a healthy cytoplasmic environment to the nDNA of an affected oocyte could offer a solution to other pathologies. For example, it was recently reported that MII‐NT could restore developmentally compromised mouse oocytes (Bai et al, 2020; Ogawa et al, 2020; Yamada & Egli, 2017) and failed fertilization after artificially activated assisted ICSI (Tang et al, 2022). The first baby born using this technology was reported by J. Zhang in 2017.…”

Section: Nt For Restoring the Developmental Competence Of Gametes And...mentioning

confidence: 99%

State of the art of nuclear transfer technologies for assisting mammalian reproduction

Gambini

Briski

Canel

2022

Molecular Reproduction Devel

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The transfer of nuclear genomic DNA from a cell to a previously enucleated oocyte or zygote constitutes one of the main tools for studying epigenetic reprogramming, nucleus-cytoplasm compatibility, pluripotency state, and for genetic preservation or edition in animals. More than 50 years ago, the first experiences in nuclear transfer began to reveal that factors stored in the cytoplasm of oocytes could reprogram the nucleus of another cell and support the development of an embryo with new genetic information. Furthermore, when the nuclear donor cell is an oocyte, egg, or a zygote, the implementation of these technologies acquires clinical relevance for patients with repeated failures in ART associated with poor oocyte quality or mitochondrial dysfunctions. This review describes the current state, scope, and future perspectives of nuclear transfer techniques currently available for assisting mammal reproduction.

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Human germline nuclear transfer to overcome mitochondrial disease and failed fertilization after ICSI

Cited by 14 publications

References 55 publications

Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles

Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles

Assisted gamete treatment to pinpoint acquired meiotic maturity and overcome oocyte activation deficiency contributed by both gametes

State of the art of nuclear transfer technologies for assisting mammalian reproduction

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