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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
School of BioSciences, Faculty of Science, Bio21 Institute, The University of Melbourne, Parkville, Victoria, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Precision Medicine Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, New South Wales, Australia
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Abstract
Seminal fluid extracellular vesicles (SFEVs) have previously been shown to interact with spermatozoa and influence their fertilisation capacity. Here, we sought to extend these studies by exploring the functional consequences of SFEV interactions with human spermatozoa. SFEVs were isolated from the seminal fluid of normozoospermic donors prior to assessing the kinetics of sperm-SFEV binding in vitro, as well as the effects of these interactions on sperm capacitation, acrosomal exocytosis, and motility profile. Biotin-labelled SFEV proteins were transferred primarily to the flagellum of spermatozoa within minutes of co-incubation, although additional foci of SFEV biotinylated proteins also labelled the mid-piece and head domain. Functional analyses of high-quality spermatozoa collected following liquefaction revealed that SFEVs did not influence sperm motility during incubation at pH 5, yet SFEVs induced subtle increases in total and progressive motility in sperm incubated with SFEVs at pH 7. Additional investigation of sperm motility kinematic parameters revealed that SFEVs significantly decreased beat cross frequency and increased distance straight line, linearity, straightness, straight line velocity, and wobble. SFEVs did not influence sperm capacitation status or the ability of sperm to undergo acrosomal exocytosis. Functional assessment of both high- and low-quality spermatozoa collected prior to liquefaction showed limited SFEV influence, with these vesicles inducing only subtle decreases in beat cross frequency in spermatozoa of both groups. These findings raise the prospect that, aside from subtle effects on sperm motility, the encapsulated SFEV cargo may be destined for physiological targets other than the male germline, notably the female reproductive tract.
Lay Summary
A male’s influence over the biological processes of pregnancy extends beyond the provision of sperm. Molecular signals present in the ejaculate can influence the likelihood of pregnancy and healthy pregnancy progression, but the identity and function of these signals remain unclear. In this study, we wanted to understand if nano-sized particles present in the male ejaculate, called seminal fluid extracellular vesicles, can assist sperm in traversing the female reproductive tract to access the egg. To explore this, we isolated seminal fluid extracellular vesicles from human semen and incubated them with sperm. Our data showed that seminal fluid extracellular vesicles act to transfer molecular information to sperm, but this resulted in only subtle changes to the movement of sperm.
Graphical abstract
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Assisted Conception Unit, Ninewells Hospital, Dundee, UK
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Infertility is estimated to affect more than 50 million couples around the world, with the male factor accounting for half of these cases, yet there is a notable absence of effective treatment options for men, other than in-vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI). This review considers unlicensed and empirical treatments used for male subfertility, including hormonal therapy, phosphodiesterase inhibitors, and antioxidants. Compounds generally demonstrate variable improvements in sperm function, but benefits for fertility are less clear. There is a pressing need for effective treatment options for subfertile men; however, our knowledge of sperm function is limited, restricting the identification of precise treatment targets. The traditional drug discovery pathway is also notorious for its extensive resource and time requirements, often extending over decades and demanding significant financial investment. Unfortunately, a substantial number of potential therapies fail before reaching the marketplace. Furthermore, reliance on mammalian models is not possible in the drug development process for male subfertility, due to significant variability between animals and man. We review recent breakthroughs and highlight novel methods aimed at improving the effectiveness and efficiency of drug discovery for male subfertility. High-throughput screening, combinatorial chemistry, and the repurposing of established medications have great potential. These strategies offer the promise of accelerating the pace of drug development, curbing the extensive demand for resources, and, in the case of drug repurposing, diminishing the demand for comprehensive pharmacokinetic and pharmacodynamic studies. As these innovative approaches are adopted, the feasibility of addressing male subfertility through scientific advancements appears to be increasingly attainable.
Lay summary
Globally, millions of men suffer from infertility, but with very few exceptions, there is no effective treatment or cure other than complex fertility treatments such as in vitro fertilisation. This review article considers various approaches to treat men, or sperm, which have been largely unsuccessful to date. A significant issue is that we lack a detailed understanding of how sperm work and why treatments do not work. Another issue is that drug discovery is slow and costly because many potential treatments fail early on. However, we discuss recent technical and technological advancements that offer hope, such as high-throughput and phenotypic screening, which allow for the rapid testing of thousands of chemicals to observe their effects on sperm, and combinatorial chemistry, which involves generating large numbers of compounds to find those with beneficial properties. Finding relevant new uses for prescription drugs could also significantly speed up the drug discovery process and result in much-needed new treatments for male infertility.
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Choline is a known developmental programming agent of the bovine preimplantation embryo. Culture of the embryo with 1.8 mmol/L choline, a concentration much higher than in blood, alters development to cause increased weaning weight and other changes during the postnatal period. It was hypothesized here that choline exerts similar effects on the developmental program of the embryo when added at concentrations similar to those in peripheral blood (i.e., 4 mol/L). Oocytes were collected via ovum pick up and embryos were produced in vitro. Embryos were cultured until day 7 after fertilization in medium with 4 mol/L choline chloride, or, as a vehicle control, with an additional 4 mol/L sodium chloride. Blastocysts were transferred into recipients and pregnancy was diagnosed at approximately 28 d of gestation. Subsequent calves (n=37 for vehicle and n=35 for choline) were weighed at birth and at weaning. Addition of choline to culture medium did not affect the proportion of embryos that became blastocysts or the proportion of transferred blastocysts that produced a pregnancy. Birth weight was unaffected by treatment but calves derived from choline-treated embryos were heavier at time of weaning and gained more per day from birth until weaning than calves derived from embryos treated with vehicle. Results demonstrate that choline can act on the preimplantation embryo at a physiologically-relevant concentration to alter postnatal phenotype. Observations are further evidence for the importance of the first days of embryonic development for the phenotype of the resulting calf.
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Maternal gut microbiome impairment has garnered attention for its potential role in influencing neurodevelopmental outcomes in offspring, especially in situations that increase brain vulnerability such as perinatal asphyxia(PA). Maternal microbiome and fetal brain interplay emerge as a critical link between maternal health and offspring neurodevelopment. This study aims to generate a model to assess the impact of maternal dysbiosis triggered by gestational antibiotic administration and PA on offspring neurodevelopment. Wistar rats were subjected to antibiotics in drinking water from the 11th gestational day until birth. On the 6th postnatal day, pups were subjected to PA/normoxia, resulting in four experimental groups: control-normoxia, antibiotics-normoxia, control-asphyxia, and antibiotics-asphyxia. Early-life behavioral tests were conducted between postnatal days 7 and 9. The initial antimicrobial cocktail (ampicillin, vancomycin, neomycin, clindamycin, amphotericin-B) led to increased number of miscarriages, poor weight gain during pregnancy, reduced offspring weight, and changes in the maternal gut microbiome compared to control. Offspring presented impaired neurodevelopmental reflexes in both PA and antibiotic groups and increased hippocampal neuroinflammation. Due to these detrimental effects, a more pregnancy-safe antibiotic cocktail was used for a second experiment (ampicillin, vancomycin, neomycin, meropenem). This resulted in no miscarriages or pregnancy-weight loss but was still linked to gut microbiome disruption. PA impaired neurodevelopmental reflexes and increased neuroinflammation, effects amplified by antibiotic administration. These preliminary findings reveal the cumulative potential of maternal dysbiosis and PA on neurodevelopment impairment, emphasizing caution in gestational antimicrobial use. Further investigations should include offspring long-term follow-up and maternal behavior and integrate probiotics to counteract antibiotic effects.
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The aim of this study was to identify pitfalls in ovarian tissue cryopreservation protocol from referral to surgical procedure and to analyze factors associated with chemotherapy exposure of the cryopreserved tissue and decreased ovarian function in a cohort of young girls at high risk of infertility.
The study population comprised 200 girls eligible for ovarian tissue cryopreservation between 2002 and 2020 at the Children's Hospital of the University Central Hospital of Helsinki (Finland). Analyses included evaluation of the proportion of patients who underwent ovarian tissue cryopreservation, factors associated with patient selection and timing of ovarian tissue cryopreservation, and ovarian function during long-term follow-up in relation to oncological treatments. Lack of counselling was identified as the major reason for not receiving ovarian tissue cryopreservation. A longer interval from scheduling gonadotoxic therapy to cryopreservation correlated with a higher exposure to alkylating agents of the ovarian tissue. The long-term ovarian function was mainly influenced by age at the time of gonadotoxic treatment. Current selection criteria for ovarian tissue cryopreservation should be implemented in order to stratify patients at risk of infertility and timely identify those at higher risk, especially in relation to age and pubertal stage. Efforts to increase healthcare providers’ awareness and facilitate guided timing in relation to the treatment protocols are needed to guarantee early access to ovarian tissue cryopreservation for all patients at high risk of infertility.
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Introduction: The Gambia, West Africa, has made recent progress on infertility, a component of sexual and reproductive health that is lagging behind others. Since 2016, there is favourable policy environment stemming from infertility research and partnership building with national stakeholders and local civil society organisations focussing on infertility. Here, we report outcomes from a participatory workshop on infertility policy implementation in The Gambia and provide insights on setting national priorities for fertility care in resource-limited settings. Methods: We conducted a participatory workshop involving 29 participants from Gambia’s public and private health sectors. Using selected participatory group work tools, stakeholders identified and prioritised key activities within the framework of five pre-defined areas of action, including (i) creating guidelines/regulations; (ii) recording/reporting data; (iii) building public-private partnership; (iv) training health providers; and (v) raising awareness and health-seeking.
Results: 17 prioritised activities were proposed across the five action areas, according to short- medium- and long-term timeframes. Three were further prioritised from the overall pool, through group consensus. A Group Model Building activity helped to envision the complexity through elucidating links, loops, and connections between each activity and their expected outcomes. Conclusions: The participatory workshop identified actionable interventions for fertility care in The Gambia, with stakeholders setting a clear path ahead. Despite challenges, the continued engagement of Gambian policymakers, practitioners, researchers, and activists in efforts to move beyond policy creation to its implementation is essential. Improving fertility care in The Gambia and other LMICs is feasible with effective collaboration and financial support.
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RESEARCH QUESTION: This study aimed to evaluate the effectiveness of a clinical decision support tool, Opt-IVF, in achieving the following outcomes: reducing the total cumulative dosage of Gonadotropins (Gn) used during controlled ovarian stimulation cycles and reducing the repeated ultrasonogram (USG) monitoring for follicular growth monitoring without compromising the number of good quality blastocysts obtained.
DESIGN: The study design employed a Multi-center Randomized Trial. The study enrolled 115 women aged 25–45 years undergoing IVF. Among the participants, 55 were randomly assigned to the intervention group (Opt-IVF), and 60 were randomly assigned to the control group. The intervention involved using a clinical decision support tool, Opt-IVF, to guide Gonadotropin dosing and trigger dates for a personalized controlled ovarian stimulation cycle.
RESULTS: The participants in the intervention group required significantly lower cumulative gonadotropin dosage during their controlled ovarian stimulation cycles. The intervention group had higher numbers of oocytes retrieved and M2 oocytes retrieved than the control group. The number of good quality blastocysts, the good blastocyst rate, the ovarian sensitivity index (OSI), and the pregnancy rate in the intervention group were significantly higher than in the control group.
CONCLUSIONS: The utilization of the clinical decision support tool led to several positive outcomes, including eliminating the need for ultrasound exams after day 5, reducing the dosage of gonadotropin required, and yielding significantly higher numbers of high-quality blastocysts and higher pregnancy rates. Thus, Opt-IVF can successfully provide a personalized, optimized, and simplified approach to superovulation. Opt-IVF consistently outperformed the clinical teams in all outcomes.
Robinson Research Institute, The University of Adelaide, Adelaide, South Australia
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Robinson Research Institute, The University of Adelaide, Adelaide, South Australia
Freemasons Centre for Male Health and Wellbeing, The University of Adelaide, Adelaide, South Australia
Genea Pty Ltd, Sydney, New South Wales, Australia
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Robinson Research Institute, The University of Adelaide, Adelaide, South Australia
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The present study determined whether adding granulocyte–macrophage colony-stimulating factor (GM-CSF) during in vitro oocyte maturation (IVM) could improve oocyte developmental competence by examining embryo development and implantation and birth rates following embryo transfer in mice. In an initial dose-response experiment, we demonstrated that the addition of 2 and 10 ng/mL GM-CSF during IVM increased cumulus expansion (P < 0.05) but did not affect fertilisation rate compared with the control group. The addition of 10 ng/mL increased blastocyst rate (17.0%; P < 0.05) and tended to increase the number of good quality blastocysts present at 96 h of culture (+19.4%; P = 0.06) and increased blastocyst inner cell mass (+25.2%; P < 0.001), trophectoderm (+29.9%; P < 0.01), and total cell numbers (+28.6%; P < 0.05). GM-CSF also reduced the incidence of DNA damage in blastocysts in the 10 ng/mL group (−16.2%) compared with the control group. These improvements translated into increases in implantation rate (+21.0%; P < 0.05) and birth rate (+17.0%; P < 0.001) following the transfer of vitrified blastocysts. GM-CSF treatment did not alter any fetal and placental parameters. Together these results suggest that the addition of GM-CSF during IVM may improve livestock in vitro embryo production and human IVM.
Lay summary
The ability to collect immature eggs from the ovaries and mature these in the laboratory is an important technology for treating certain types of infertility in women as well as for preserving their fertility, for example prior to cancer treatment. This technique is called in vitro oocyte maturation or IVM and is also used in animal breeding. However, pregnancy and birth rates in humans and animals using this technique are lower than that which can be achieved using natural mating. We have shown that adding GM-CSF, a molecule found in the ovary, during IVM can increase the number and quality of embryos produced in mice. We have also found that when these embryos are transferred to surrogate mothers, implantation and birth rates are increased. These results suggest that the addition of GM-CSF during IVM may improve pregnancy and birth rates in humans as well as animals.
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First-trimester pregnancy losses are commonly attributed to chromosomal abnormalities. The causes of pregnancy loss following transfer of a euploid embryo are not fully elucidated. The aim of this study was to evaluate clinical and embryological parameters for pregnancy failure following the transfer of a single euploid embryo. Pregnancy outcomes of single euploid embryo transfers from a single centre between January 2017 and March 2020 were retrospectively evaluated. Several clinical and embryological parameters were evaluated in consideration to pregnancy outcomes; total pregnancy loss and live birth (LB). Endometrial preparation type, number of previous frozen embryo transfer cycles, history of recurrent pregnancy loss, higher body mass index, presence of endometriosis and/or adenomyosis and embryo quality were found to be significantly different between two groups. Morphokinetic parameter analysis of 523 euploid embryos using time-lapse imaging did not show any statistical differences between the two groups; however, a significantly higher rate of uneven blastomeres in the cleavage stage was observed in the total pregnancy loss group. Evaluation of clinical and embryological data can reveal possible factors associated with pregnancy loss that can facilitate improved patient consultation. Feasible interventions can potentially increase the chance of achieving an LB.
Lay Abstract
Like natural pregnancies, not all pregnancies following fertility treatment go to term. The most common reason for these losses is that these embryos lack the genetic constitution compatible with live birth. Combined with fertility treatment, genetic tests can evaluate the genetic ability of embryos to go to term. Monitoring the outcome of pregnancies resulting from such embryos can help us identify whether and which conditions specific to treatment can lead to pregnancy loss. The analysis identified four parameters associated with embryo loss: Embryo quality and division patterns, existence of previous treatment and treatment type.
Department of Obstetrics and Gynaecology, Monash University, Melbourne, Clayton, Victoria, Australia
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Wellcome Centre for Human Genetics, University of Oxford, Oxford, United Kingdom
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Wellcome Centre for Human Genetics, University of Oxford, Oxford, United Kingdom
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Graphical abstract
Abstract
Immunological dysregulation plays a fundamental role in the inflammatory aspects of endometriosis. Circulating blood leukocytes, one of the most abundant immune cell populations in the human body, have been shown diagnostic significance in some diseases. Nevertheless, the association between peripheral blood leukocyte counts and endometriosis remains unexplored to date.
We analyzed two targeted study cohorts: a tertiary center cohort (Endometriosis at Oxford University (ENDOX) Study: 325 cases/177 controls) and a large-scale population study (UK Biobank (UKBB): 1537 cases/6331 controls). In both datasets, peripheral venous blood sample results were retrieved, and counts of leukocyte subpopulations, including neutrophils, lymphocytes, monocytes, eosinophils, and basophils, were analyzed. Logistic regression models were used to investigate the association of leukocyte subtype alterations with endometriosis status, adjusting for confounding factors. We demonstrate that a higher blood basophil level is associated with increased odds of endometriosis. This association was first discovered in the ENDOX cohort (basophils >0.04 × 109/L: OR 1.65 (95% CI: 1.06–2.57), P trend = 0.025) and replicated in the UKBB dataset (basophils >0.04 × 109/L: OR 1.26 (95% CI: 1.09–1.45), P trend = 0.001). Notably, women with basophil counts in the upper tercile had significantly increased odds of having stage III/IV endometriosis (ENDOX study: OR = 2.30, 95% CI (1.25–4.22), P trend = 0.007; UKBB study (OR = 1.40, 95% CI (1.07–1.85), P trend = 0.015). None of the other leukocyte subtypes showed an association. Our findings suggest an association between inflammatory responses and the pathogenesis of endometriosis; future studies are warranted to investigate whether the association is causal.
Lay summary
Endometriosis is a long-term disease affecting approximately 10% of women during their fertile age. It happens when the tissue similar to the lining of the womb grows in other parts of the body, commonly causing pelvic pain and subfertility. Most diagnostic tests for endometriosis are neither accurate nor reliable, leading to a long wait before a correct diagnosis. Looking for changes in blood cell counts could guide doctors for further testing to confirm diagnosis. Our study shows that a higher number of basophils, a specialized type of white cells, commonly measured in a simple blood test, are positively linked with a higher likelihood of endometriosis. The link becomes stronger in severe endometriosis cases. Although we are showing a robust link, whether this can be used to find endometriosis sooner needs to be tested in future studies.
