Systems Biology in Reproductive Medicine最新文献

Human fetal development requires sustenance via the placenta, which mediates molecular transport between maternal and fetal circulations. Placental formation begins as cells of the trophoblast lineage differentiate and the extraembryonic mesoderm becomes vascularized, assembling a unique organ de novo that facilitates nutrient and gas exchange, waste removal, hormone production and immune modulation. We describe how placentation is orchestrated to keep pace with fetal growth, but is vulnerable to disruption by medical interventions for infertility. Initially, trophoblast stem cells differentiate into proliferating mononuclear cytotrophoblasts (CTBs) that fuse to form the multinucleated syncytiotrophoblast (STB). The STB ensheathes the chorionic villi, bathed in maternal blood. As fetal blood vessels develop within the mesodermal core of villi, the maternal-fetal interface is established. Where the villi meet the decidua, CTBs further differentiate into extravillous trophoblasts, which invade and remodel uterine arteries into high-conductance, low-resistance vessels, enhancing maternal blood flow to the placenta. Among the critical intercellular axes that govern trophoblast differentiation, invasion, and vascular remodeling hormonal cues, particularly those associated with the corpus luteum, are critical; their alteration in certain assisted reproductive technology (ART) protocols can contribute to incomplete arterial remodeling. Malplacentation is linked to miscarriage, fetal growth restriction, and preeclampsia, affecting over 10% of pregnancies, and occurring at higher rates in patients diagnosed with infertility, especially those who conceive through ART. Understanding the mechanisms driving these pathologies is essential for improving pregnancy outcomes. Strategies to optimize ART protocols and therapeutic interventions targeting key signaling pathways offer potential avenues to mitigate risks associated with malplacentation.

Recent evidence suggests that individuals with polycystic ovary syndrome (PCOS) have an increased risk of developing mental health disorders and comorbidities linked to nervous system dysfunction. Interestingly, patients with schizophrenia (SCZ) often exhibit PCOS symptoms, indicating a possible connection between the two conditions. However, the underlying molecular links between these diseases remain poorly understood. We employed a comprehensive in-silico approach, utilizing publicly available datasets to investigate shared biomarkers candidates and key regulators involved in the development of PCOS and SCZ. We retrieved the datasets from the NCBI GEO database and differentially expressed genes (DEGs) were identified for each dataset. Common DEGs (cDEGs) were determined, and transcription factors (TFs) and miRNA targeting cDEGs were examined using the mirDIP portal and TRRUST database, respectively. We also assessed the TF-miRNA interactions by TransmiR database and constructed a regulatory network including TFs-microRNAs-cDEGs. Our analysis identified a total of 15 cDEGs that are regulated by 15 TFs and 8 mRNAs. Among our findings, we prioritized RELA as a potential TF regulator for both diseases, demonstrating synergistic interaction with four cDEGs (EGR1, CXCL8, IL1RN, IL1B) and seven microRNAs (hsa-miR-580, hsa-miR-5695, hsa-miR-936, hsa-miR-3675, hsa-miR-634, hsa-miR-603, hsa-miR-222) that target these genes. Our data highlights potential common biomarkers for PCOS and SCZ, presenting a novel regulatory network that elucidates the molecular mechanisms underlying both conditions. This emphasizes the importance of further research to explore new translational approaches, which may ultimately lead to improved diagnostic and therapeutic strategies for affected individuals.

Infertility and low birth rates have been threatening developed countries and have become a major socioeconomic problem that requires attention. To this end, germplasm preservation and subsequent fertility restoration with the aid of various assisted reproductive technologies (ART) have overcome many infertility issues over the last four decades. Particularly, cryopreservation offered various fertility preservation (FP) options for people with reproductive issues, gender dysphoria, and significantly expanded reproductive lifespan. The aim of this review is to provide the reader with the role that cryopreservation has played in human reproductive medicine (HRM) and family planning. We will first discuss the fundamentals of cryopreservation, including the development of cryopreservation methodologies for human reproductive cells and tissues, and how these techniques have helped address human infertility problems, as well as other ethical, legal, and social challenges. We will focus on literature regarding the current methodologies used in human infertility laboratories' clinical setting of the oocyte, sperm, and embryo, gonad cryopreservation, and discuss the latest issues related to the implementation of cryopreservation strategies used in HRM.

The objective of this study was to evaluate the effect of a tannin-rich extract from commercial Acacia mearnsii (MTE_0), and gallocatechin, a flavonoid compound derived from Acacia mearnsii, on the long-term viability and motility of cryopreserved ovine semen. Six fresh ejaculates obtained from six adult merino rams twice per week for three weeks were allocated to five aliquots (0, 12.5, 25, 50, and 100 µM gallocatechin added into the Optidyl™ extender) before cooling and cryopreservation. Effects of MTE_0 and gallocatechin on post-thawed motility characteristics were analyzed using computer-assisted semen analysis (CASA), and viability (LIVE/DEAD^® kit, Molecular Invitrogen, Waltham, MA), oxidative stress (2,7-dichlorodihydrofluorescein diacetate (H2DCFDA, Thermo Fisher^®, Waltham, MA)) for reactive oxygen species (ROS), mitochondrial membrane potential (JC-1 MitoTracker, Molecular Invitrogen, Waltham, MA), acrosomal integrity (lectin PNA), and capacitation using merocyanine 540 (M540) and YO-PRO-1 dyes in flow cytometry. Data were analyzed using one-way ANOVA (IBM SPSS 21.0 for Windows, Armonk, NY). Gallocatechin at 25 µM positively affected (p ≤ .001) kinematic parameters including average path velocity (VAP), progressive velocity (VSL), and beat cross frequency (BCF) of cryopreserved semen. Similarly, gallocatechin at 25 µM^- improved sperm motility (live 21.99 ± 2.06%), reduced ROS levels (26.45 ± 1.10%), and mitigated premature capacitation (viable and stable 20.08 ± 1.48%) compared to other treatments. Gallocatechin addition to semen resulted in a significant (p ≤ .001) positive effect compared with the MTE_0 extract. It is concluded that gallocatechin inclusion at 25 µM significantly reduces semen deterioration following cryopreservation. This study is the first to introduce gallocatechin as an efficient antioxidant additive to ovine semen to improve its quality during storage. Our findings will help improve post-thaw ovine semen quality and longevity. Future studies to elucidate the mechanism of anti-oxidative stress action of gallocatechin and its derivatives on semen motility and longevity are recommended.

Components of the renin-angiotensin system (RAS) are expressed in both female and male reproductive tracts, with angiotensin I converting enzyme (ACE) being an important component for male reproductive function, as shown in animal models. The most studied ACE polymorphism is the Alu insertion-deletion (I/D), which has been proposed to have a negative effect on male fertility. Given the conflicting evidence in the literature, we conducted a multicentric case-control study to investigate the association between the ACE Alu I/D polymorphism and impaired spermatogenesis. Using PCR amplification and agarose electrophoresis, we genotyped the ACE gene Alu I/D polymorphism in 745 South Slavic men. The study group consisted of 457 patients with impaired spermatogenesis, 239 with non-obstructive azoospermia (NOA) and 218 with oligoasthenoteratozoospermia (OAT) and a control group of 288 fertile men. No association was found between the Alu I/D polymorphism and these semen phenotypes, suggesting that it is not associated with NOA or severe OAT in this cohort. To provide a broader regulatory context, we also developed an integrative atlas of ACE regulatory elements by in silico multi-omics analysis using genomics databases and bioinformatics tools. Data integration revealed various regulatory mechanisms at multiple omics levels, including genomics, epigenomics, miRNAomics, transcriptomics, proteomics and epiproteomics. These include genomic variants with predicted deleterious effects, a CpG island, microRNAs (miRNAs) and post-translational modifications (PTMs). In addition, protein interaction analysis revealed that ACE is indirectly linked to several proteins previously associated with male infertility and is also targeted by miRNA previously associated with oligozoospermia. This comprehensive, multi-faceted approach, combining genetic association analysis with bioinformatics, provides insights into ACE regulation in its broader molecular context. These results emphasize the importance of further integrative multi-omics and systems biology research to better understand the role of ACE in male reproductive function.

Cryopreservation of testicular and epididymal spermatozoa is more challenging in comparison to ejaculated spermatozoa due to lower sperm concentration and motility, and higher sperm sensitivity to cryoprotectants. Sperm vitrification without the use of potentially toxic permeable cryoprotectants is an attractive freezing alternative for testicular and epididymal spermatozoa, as well as oligoasthenoteratozoospermia (OAT) samples. Our study is a retrospective analysis of outcomes in IVF cycles involving a total of 70 testicular, 77 epididymal and 69 ejaculated OAT samples vitrified in a closed double-straw device using mHTF medium supplemented with protein and sucrose, without any permeable cryoprotectant. In total, 71 frozen samples were used for intracytoplasmic sperm injection (ICSI). Results were compared to fresh samples (26 testicular, 53 epididymal and 63 ejaculated OAT samples) that served as controls. Elective single frozen embryo transfers of euploid or unknown-ploidy blastocysts were performed. While sperm motility is expected to diminish following slow sperm freezing and thawing, our data demonstrated that vitrification of testicular, epididymal and OAT samples had a mean motility rate comparable to fresh samples. No statistically significant differences (p > 0.05) were observed between vitrified versus fresh TESE in fertilization (64.1% vs. 59.5%), blastocyst development (54.9% vs. 56.7%), blastocyst euploidy (36.4% vs. 33.3%), clinical pregnancy (47.8% vs. 36.4%) and live birth rates (43.5% vs. 24.2%). Similarly, vitrified versus fresh PESA showed no statistically significant differences (p > 0.05) in the analyzed results respectively: (69.4% vs.74.9%; 62.6% vs. 59.7%; 40.5% vs. 48.1%; 36.0% vs.37.7%; and 32.0% vs. 27.5%). For vitrified OAT samples, there was a significant difference in blastocyst development and euploidy rates when compared to the control group. Our results demonstrate that human testicular, epididymal spermatozoa and samples with OAT can be successfully vitrified in small volumes in a closed system without using any permeable cryoprotectants, allowing utilization of this technique in clinical settings.

The variation in reproductive age among individuals is significant, with many cases of infertility involving premature ovarian aging. This issue, combined with the societal trend of delaying childbearing, leads to age-related ovarian dysfunction. Ovarian aging is related to a decline of ovarian reserve, as oocyte quantity, quality, and precocious senescence, and may affect fertility and the overall individual well-being. Mitochondria play a central role in the maintenance of any cell health. Then mitochondrial dysfunctions may be responsible also for a negative impact on the quality, number, and function of oocytes, leading to different age-related reproductive disorders, impaired oogenesis, and embryogenesis. Although a large number of researches have shown clearly that mitochondrial dysfunction and morphology changes affect the maintenance and function of all major organs and tissues, such as the brain, heart, skeletal muscle, liver, and others the mechanisms contributing to early ovarian aging, a decrease of oocyte quality, and infertility remain unclear. In this review, we summarize the role of mitochondrial dysfunction in ovarian aging, presenting recent findings on morpho-functional changes in these organelles, and highlighting how their dysfunction accelerates ovary and cell senescence. We also explore their impact on oocyte functions. The reported data highlight the critical role of mitochondria in maintaining and enhancing oocyte quality, indicating that future studies should further focus on the mechanisms underlying mitochondrial damage and on identifying mitochondrial targets that may offer promising strategies to preserve, recover, and extend fertility in aging women.

Genetic variants are known causes of male infertility and oligoasthenoteratozoospermia (OAT), as shown by knockout mouse models and patients with infertility. However, most OAT cases lack a definitive genetic diagnosis. Peripheral blood and semen samples were collected from a patient with OAT. Semen analysis, Papanicolaou staining, transmission electron microscopy, whole-exome sequencing (WES), Sanger sequencing, and in silico analyses, such as conservative analysis and conformational analyses, were used to investigate the genetic causes of OAT. Semen analysis revealed a notable reduction in sperm count and motility, and defects in sperm morphology. Light and electron microscopy showed numerous defects in the head-to-tail coupling apparatus of the sperm, and a small number of structural defects in the sperm flagella. WES identified two novel homozygous missense variants SUN5: c.G703A/p.A235T and DNAH10: c.A1436G/p.Q479R. The p.A235T and p.Q479R variants were predicted to generate aberrant SUN5 and DNAH10 proteins using in silico analysis. Here, we report the identification of two novel deleterious variants of SUN5 and DNAH10 associated with OAT, expanding the mutant spectrum of male infertility.

Early embryo development and competence mechanisms are paramount to ART's success but are still underexplored in human-relevant animal models. Clinical embryo evaluation remains largely based on subjectively evaluated morphological characteristics. In the current era of biomarkers and low-input high-throughput technologies, it is possible to investigate the multi-complex molecular landscape of the embryo during development from the zygote up to the blastocyst stage. The scope of this article is to review the underlying molecular mechanisms of pre-implantation embryo development. In addition, we briefly explore the latest advances in embryo quality assessment - including molecular and epigenetic evaluation and technical advances such as time-lapse imaging and artificial intelligence - together with clinical challenges for selecting the best embryo for single embryo transfer.

Ovarian stimulation is applied to women undergoing IVF treatment with the aim of selecting more than one follicle. Until a few years ago, fresh embryo transfer was the norm, but not without the risk of ovarian hyperstimulation syndrome (OHSS). In recent years, vitrification has allowed the successful freezing of all oocytes or embryos, thus favoring multiple embryo transfer in subsequent thawing cycles and eliminating the risk of OHSS. Certainly, excess embryos from fresh embryo transfer cycles can be also frozen and thawed in future transfers. In terms of the number of oocytes needed, research in recent years has shown differences between fresh embryo transfer cycles and elective freezing of all embryos, termed elective freeze-all cycles, with the fresh cycle requiring an optimal number to avoid OHSS, while in elective freeze-all cycles there appear to be no specific upper limit. Consequently, the approach to ovarian stimulation may also differ between the two types of cycles. Although GnRH antagonists are used in both cycles, in elective freezing cycles progestins tend to replace antagonists in preventing the endogenous LH surge. However, it is unclear whether prevention of the LH surge is required in such cycles, since luteinization affecting the endometrium is not clinically relevant, as no embryo transfer occurs. This narrative review describes the current experience in ovarian stimulation for IVF, highlighting the differences between fresh and elective freeze-all cycles and the potentially different approach.