Reproduction and Fertility

Age and serum anti-Müllerian hormone levels as predictors of time to return of menses after chemotherapy

AbstractChemotherapeutic agents result in the loss of growing follicles, which can manifest as amenorrhoea. Alkylating chemotherapy (AC) is known to be more gonadotoxic than non-alkylating chemotherapy (NAC). Anti-Müllerian hormone (AMH), an indirect marker of ovarian reserve, and age have been investigated as predictors of ovarian function after chemotherapy; however, little is known about the time to return of menses. This study aimed to assess how patient age and baseline serum AMH levels at cancer diagnosis affect the time to return of menses post-chemotherapy. This retrospective cohort study examined oncology patients (n = 67) who underwent chemotherapy and were treated through the Reproductive Services Unit of two institutions in Melbourne, Australia. Primary outcomes included the correlation between age and baseline AMH with time to return of menses after chemotherapy. Secondary outcomes include the change in AMH levels at 6- and 12-months post-completion of chemotherapy. Pairwise correlation of the pre-chemotherapy AMH level and time to return of menses demonstrated statistical significance (Spearman’s coefficient, ρ = −0.40) for patients who underwent AC. This analysis in breast cancer patients who underwent AC displayed a negative correlation but was not statistically significant. No association was found between age and time to return of menses for all cancer (NAC or AC) or breast cancer patients who underwent AC. Higher AMH levels prior to AC were associated with an earlier return of menses after chemotherapy. Age at the commencement of chemotherapy was not associated with return of menses. Further prospective research is required to assess post-chemotherapy recovery of AMH.Lay summaryChemotherapy, used to treat cancer, is known to damage women’s ovaries, with certain types having a more toxic effect than others. This may result in a temporary loss of periods while undergoing chemotherapy. AMH is a hormone produced by the ovaries and gives an indication of their level of function. This study looks at whether an individual's AMH or age when beginning chemotherapy can predict the time before the resumption of periods after completing chemotherapy. This study found that for cancer patients who underwent the chemotherapy type known to be more toxic to ovaries, the higher their AMH level was before beginning chemotherapy, the more rapidly their periods would return after completing chemotherapy. Age was not found to accurately predict how rapidly periods would return after completing chemotherapy. This information can be used to inform patients before treatment of the chances of periods returning and, consequentially, pregnancy after the completion of their chemotherapy.

AZFa candidate gene UTY and its X homologue UTX are expressed in human germ cells

The Ubiquitous Transcribed Y (UTY a.k.a. KDM6C) AZFa candidate gene on the human Y chromosome and its paralog on the X chromosome, UTX (a.k.a. KDM6A), encode a histone lysine demethylase removing chromatin H3K27 methylation marks at genes transcriptional start sites for activation. Both proteins harbour the conserved Jumonji C (JmjC) domain, functional in chromatin metabolism, and an extended N-terminal tetratricopeptide repeat (TPR) block involved in specific protein interactions. Specific antisera for human UTY and UTX proteins were developed to distinguish the expression of both proteins in human germ cells by immunohistochemical experiments on appropriate tissue sections. In the male germ line, UTY was expressed in the fraction of A spermatogonia located at the basal membrane, probably including spermatogonia stem cells. UTX expression was more spread in all spermatogonia and in early spermatids. In female germ line, UTX expression was found in the primordial germ cells of the ovary. UTY was also expressed during fetal male germ cell development, whereas UTX expression was visible only at distinct gestation weeks. Based on these results and the conserved neighboured location of UTY and DDX3Y in Yq11 found in mammals of distinct lineages, we conclude that UTY, such as DDX3Y, is part of the Azoospermia factor a (AZFa) locus functioning in human spermatogonia to support the balance of their proliferation-differentiation rate before meiosis. Comparable UTY and DDX3Y expression was also found in gonadoblastoma and dysgerminoma cells found in germ cell nests of the dysgenetic gonads of individuals with disorders of sexual development and a Y chromosome in karyotype (DSD-XY). This confirms that AZFa overlaps with GBY, the Gonadoblastoma susceptibility Y locus, and includes the UTY gene. Lay Summary AZFa Y genes are involved in human male germ cells development and support gonadoblastoma (germ cell tumour precursor cells) in the aberrant germ cells of the gonads of females with genetic disorders of sexual development. The AZFa UTY gene on the male Y chromosome is equivalent to UTX on the female X chromosome. These genes are involved in removing gene regulators to enable activation of other genes (i.e. removal of histone methylation known as epigenetic modifications). We wanted to learn the function of UTY and UTX in developing sperm and eggs in human tissues and developed specific antibodies to detect both proteins made by these genes. Both UTY and UTX proteins were detected in adult and fetal sperm precursor cells (spermatogonia). UTX was detected in egg precursor cells (primordial germ cells). UTY was detected in gonadoblastoma and dysgerminoma tumour cells (germ cell tumours originating from genetic disorders of sexual development due to having a Y chromosome). Based on our study, we conclude that UTY is not only part of AZFa, but also of GBY the overlapping gonadoblastoma susceptibility Y region.

Matrix metalloproteinase-induced cervical extracellular matrix remodelling in pregnancy and cervical cancer

Graphical abstract Abstract The phenomenal extracellular matrix (ECM) remodelling of the cervix that precedes the myometrial contraction of labour at term or preterm appears to share some common mechanisms with the occurrence, growth, invasion and metastasis of cervical carcinoma. Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that are pivotal to the complex extracellular tissue modulation that includes degradation, remodelling and exchange of ECM components, which contribute to homeostasis under normal physiological conditions such as cervical remodelling during pregnancy and puerperium. However, in cancer such as that of the uterine cervix, this extensive network of extracellular tissue modulation is altered leading to disrupted cell–cell and cell–basement membrane adhesion, abnormal tissue growth, neovascularization and metastasis that disrupt homeostasis. Cervical ECM remodelling during pregnancy and puerperium could be a physiological albeit benign neoplasm. In this review, we examined the pathophysiologic differences and similarities in the role of MMPs in cervical remodelling and cervical carcinoma. Lay summary During pregnancy and childbirth, the cervix, which is the barrel-shaped lower portion of the womb that connects to the vagina, gradually softens, shortens and opens to allow birth of the baby. This process requires structural and biochemical changes in the cervix that are stimulated by enzymes known as matrix metalloproteinases. Interestingly, these enzymes also affect the structural and biochemical framework of the cervix during cervical cancer, although cervical cancers usually occur after infection by human papillomavirus. This review is intended to identify and explain the similarities and differences between the structural and chemical changes in the cervix during pregnancy and childbirth and the changes seen in cervical cancer.

The impact of hypoxia on the cellular phenotype and proteome of KGN cells

Graphical Abstract Abstract Human KGN cells are a widely studied cellular model for human granulosa cell tumors (GCTs), as well as for non-tumorous human granulosa cells. Such studies are typically performed at oxygen (O 2 ) levels that are equivalent to the atmospheric concentration for practical reasons. However, in most tissues of the human body, as well as in tumors, much lower O 2 levels exist. Hypoxic conditions can regulate the phenotype and the behavior of cells in a cell-type-specific way. However, how they may impact KGN cells was not well known. The aim of this study was therefore to evaluate the consequences of a defined hypoxic condition (1% O 2 ; 10 days) in KGN cells. Compared to cells cultured in atmospheric conditions, hypoxia reduced the numbers and increased the size of KGN cells but did not alter cell velocity in cell migration studies. Hypoxia affected the cellular proteome of KGN cells (57 more and 75 less abundant proteins). The associated pathways indicate, among others, metabolic adaptations and mitochondrial changes evoked by hypoxia. The cellular responses may be related, in part, to the increased nuclear expression of hypoxia-inducible factor 1α (HIF1α). The results provide a detailed picture of hypoxia-induced changes in KGN cells and are a resource for future studies. If transferable to the in situ situation, they shed light on how variable O 2 levels within the GCT microenvironment may regulate tumor cells and thereby may contribute to tumor cell heterogeneity. Lay summary Human cells can be isolated from the body and kept alive in incubators, in which, mainly for practical reasons, the oxygen (O 2 ) levels typically correspond to the surrounding atmosphere. This does not reflect the conditions within the human body, where much lower O 2 levels exist. This study shows the importance of low O 2 levels on the cellular make-up and behavior of KGN cells, which are accepted models for a group of ovarian tumors (GCTs), as well as normal, non-tumorous granulosa cells (from which GCTs originate). We found that when KGN cells were exposed to low O 2 levels, cell numbers and cell size, as well as cellular proteins and RNA, changed. The experiments, revealing adaptations associated with hypoxia, may contribute to a better understanding of the normal ovary and ovarian tumors.

The stem bark decoction of Myrianthus arboreus P. Beauv. (Cecropiaceae) shows anti-uterine leiomyoma effects in Wistar rat

Graphical abstract Abstract Over the past 30 years, the number of new cases of uterine leiomyomas (UL) in women of reproductive age has increased by 67.14% worldwide. The limitations of the current therapeutic options have led to the search for alternatives. Myrianthus arboreus P. Beauv., used for infertility and tumors, has never been tested for UL. In the present study, the decoction of its stem bark was evaluated in a model of UL induced in female Wistar rats. Animals were treated once daily by gavage for 30 days. Normal control and model groups received distilled water, positive groups received mifepristone (5.2 mg/kg) and the remaining three groups were treated with 50, 100, and 200 mg/kg M. arboreus extract, respectively. Compared to the model group, the extract at 50 and 100 mg/kg reduced the E2B- and progesterone-induced uterine horn asymmetry and thickening, and the relative uterine weight and diameter; myometrial thickness; and collagen density (at 100 and 200 mg/kg). Regarding cytokines, the extract decreased the uterine levels of TGF-β1 and VEGF (at 100 and 200 mg/kg) and TNF-α (at all doses tested). It also decreased the serum levels of estradiol (at 100 and 200 mg/kg). Despite positive trend in reducing oxidative damage (decreased MDA at 50 mg/kg, increased catalase activity at all tested doses, and increased GSH at 100 mg/kg), the level of oxidative stress is still elevated. By attenuating key cellular events involved in the growth and development of UL, such as inflammation, fibrosis and angiogenesis, Myrianthus arboreus may be a promising option for UL treatment and management. Lay summary Uterine fibroids are non-cancerous tumors that originate from the smooth muscular wall of the uterus. They may cause severe symptoms, such as chronic pelvic pain, heavy menstrual bleeding, painful intercourse, frequent urination, back pain and obstetric complications (embryo implantation failure, fetal growth restriction, miscarriages, preterm delivery, and fertility impairment). The exact cause is not known, but hormonal changes may play an important role. Treatment options depend on the symptoms and desire to preserve the uterus, and may include medication, minimally invasive procedures and surgery. Their associated risks and limitations fuel searching for conservative alternatives. This study investigates the anti-uterine fibroid potential of the giant yellow mulberry (Myrianthus arboreus P. Beauv.) in rat. Consuming boiled M. arboreus stem bark in water reduced uterine wall thickness, inflammation, and collagen deposition, and increased the ability of the uterus to fight cell damage. Thus, this extract may prevent or shrink uterine fibroids by reducing uterine swelling, fibrosis and excessive cell growth.