Cause Of Premature Ovarian Insufficiency Research Articles

Premature ovarian insufficiency in patients with galactosemia

Background. Galactosemia is a congenital disorder of carbohydrate metabolism caused by a defect in any of the enzymes of galactose metabolism. One of the long-term complications is premature ovarian insufficiency (POI), which is more common in patients with the c.563A>G (Q188R) mutation in the homozygous state in the galactose-1-phosphate uridylyltransferase (GALT) gene. At the same time, fertility factors may be higher in patients with POI caused by classical galactosemia (CG) compared with other causes of POI, which makes it difficult to resolve the issue of the need to use fertility preservation methods for this group of patients in the prepubescent period. Case report. This article describes two clinical observations of patients with CG who were diagnosed with hypergonadotropic hypogonadism. Patient A. was initiated hormone replacement therapy (HRT) at the age of 11, and according to the results of osteodensitometry, there is currently no decrease in bone mineral density. In patient C. at the age of 14, before the start of HRT, ovaries without pronounced follicular apparatus, osteopenia and osteoporosis were detected. The issue of the necessity and timing of the use of fertility preservation methods is being considered. Conclusion. Patients with CG are recommended to monitor hormonal profile indicators for timely administration of HRT. Cryopreservation of ovarian tissue should be considered as one of the options for maintaining fertility in patients with CG, taking into account that some of them still have the possibility of spontaneous pregnancy, despite the POI.

Human mesenchymal stem cells derived exosomes improve ovarian function in chemotherapy-induced premature ovarian insufficiency mice by inhibiting ferroptosis through Nrf2/GPX4 pathway.

Chemotherapy exposure has become a main cause of premature ovarian insufficiency (POI). This study aimed to evaluate the role and molecular mechanism of human umbilical cord mesenchymal stem cell-derived exosomes (hUMSC-Exos) in ovarian function protection after chemotherapy. hUMSC-Exos were applied to cyclophosphamide-induced premature ovarian insufficiency mice and human ovarian granulosa tumor cells (KGN) to determine their effects on follicular development and granulosa cell apoptosis. Evaluation was done for iron ion and reactive oxygen species (ROS) production, lipid peroxidation levels, and changes in iron death-related molecules (nuclear factor (erythroid-derived 2)-like 2 (Nrf2), Glutathione Peroxidase enzyme 4 (GPX4), and Solute carrier family 7 member 11 cystine glutamate transporter (SLC7A11; xCT)). Furthermore, rescue experiments using an Nrf2 inhibitor were performed to assess the therapeutic effects of hUMSC-Exos on granulosa cells. hUMSC-Exos promoted ovarian hormone levels and primary follicle development in POI mice and reduced granulosa cell apoptosis. After hUMSC-Exos treatment, the ROS production, free iron ions and lipid peroxidation levels of granulosa cells decreased, and the iron death marker proteins Nrf2, xCT and GPX4 also decreased. Furthermore, the Nrf2 inhibitor ML385 significantly attenuated the effects of hUMSC-Exos on granulosa cells. hUMSC-Exos inhibit ferroptosis and protect against CTX-induced ovarian damage and granulosa cell apoptosis through the Nrf2/GPX4 signaling pathway, revealing a novel mechanism of hUMSC-Exos in POI therapy.

Menopause and premature ovarian insufficiency in rheumatic diseases: A review

Premature ovarian insufficiency (POI) is a clinical syndrome defined as loss of ovarian function before the age of 40 years. Among the causes of POI are genetic, metabolic disorders, as well as infectious or iatrogenic factors, but in some cases the exact cause cannot be established. Given the possible association of ovarian failure with autoimmune disorders, as well as the detection of antibodies to normal ovarian tissue in the serum of patients with POI, it is assumed that an autoimmune mechanism is a significant cause of the syndrome. Taking into account the relationship between POI and autoimmune diseases, it seems relevant to address the issues of mutual influence of the ovarian function of women with POI and rheumatic diseases (in particular, systemic lupus erythematosus, rheumatoid arthritis and systemic scleroderma), which is presented in this review. Hormone replacement therapy may be recommended for women with POI to improve their quality of life.
 Keywords: premature ovarian insufficiency, menopause, anti-ovarian antibodies, systemic lupus erythematosus, rheumatoid arthritis, systemic scleroderma, rheumatic diseases, hormone replacement therapy.

FRI556 Thyroid Autoimmunity Leading To Premature Ovarian Insufficiency (POI) In A Euthyroid Young Woman

Abstract Disclosure: P. rimal: None. M. Moeed: None. C. Musurakis: None. A. Ojha: None. W. Akhtar: None. C.P. Barsano: None. M.F. Siddiqui: None. Introduction: Premature Ovarian Insufficiency is rare in young women and affects only 0.1% before the age of 30. The causes of POI in most cases remain unidentified, with autoimmunity being responsible for 4-30% of the cases. We present the case of a young euthyroid woman who presented for the evaluation of primary amenorrhea and diagnosed with POI associated with thyroid autoimmunity. Clinical Case: A 28-year-old Afghani woman with no known co-morbidities was referred to endocrinology clinic for the work up of primary amenorrhea. She was a goitrous and had normal secondary sexual characteristics and a history of withdrawal bleeding with OCP. There was no known personal or family history of autoimmune disease. Work up revealed hypergonadotropic hypogonadism with high FSH, 55 mIU/ml[Ref range for postmenopausal woman 16.7-113], high LH, 20.28 mIU/ml [Ref range for postmenopausal woman 10.8-58.69] and low estradiol, <2 pg/ml. Anti-Mullerian hormone was low, <0.08ng/ml [RefRange 0.69-13.39] signifying inadequate ovarian reserves and pelvic ultrasound confirmed atrophic ovaries. Etiological work up revealed normal female karyotype, 46XX, and negative FMR gene mutation testing for Fragile X syndrome. Thyroid functions were normal TSH, 3.3 mIU/L (0.27-4.2) and Free T4, 1.29 ng/dl[Ref Range 0.70-2.7]. Autoimmune work up was negative for adrenal 21-OH antibody and anti-ovarian antibodies. However, patient had strongly positive thyroid peroxidase (TPO) antibodies, 613.77 mIU/L[Ref range 0.0-9.0] consistent with thyroid autoimmunity. DXA scan revealed osteopenia. Thus, the diagnosis of POI associated with thyroid autoimmunity was established. HRT with estradiol patch and progesterone was initiated. Calcium, Vitamin D supplements and fertility counseling were provided. Conclusion: Thyroid disease is the most common autoimmune disease associated with POI. Studies have shown higher risk of POI and infertility in patients with Hashimoto’s and Graves’ diseases with co-existing high titers of TPO or anti-thyroglobulin antibodies. Ovaries are subject to autoimmune attack that leads to diminished ovarian reserves. However, until now the association of isolated elevation of TPO antibodies with POI has not been established. A Study by Osuka et al. did not find any significant difference in AMH levels between TPO and non-TPO positive euthyroid women and concluded that unlike hypothyroid women, thyroid autoantibodies were not likely to influence ovarian reserves in euthyroid women. Our case is unique where POI is associated with isolated elevation of TPO antibodies in the absence of clinical thyroid disease. This signifies the possibility that ovarian failure may precede the onset of thyroid disease in patients with thyroid autoimmunity and should be suspected and treated at an early stage, before ovarian reserves are diminished. Presentation: Friday, June 16, 2023

Premature ovarian insufficiency caused by the treatment of ovarian malignant mixed germ cell tumor: a case report

Background: Premature ovarian insufficiency (POI) will bring women early and long-term adverse consequences, including estrogen deficiency-related symptoms (hot flashes, sweating, decreased libido, insomnia, etc.) and significantly increases the risk of chronic diseases including Alzheimer's disease, cardiovascular disease and osteoporosis, along with increased risk of early death. More importantly, POI has greater harm to women’s fertility. Mostly the causes of POI are unknown. Most important are iatrogenic factors such as radiotherapy, chemotherapy and surgery, which increase the risk of POI.

P-690 RESEarch on ovarian reseRVE among Neurologic patients: the RESERVEN study

Abstract Study question Could Multiple Sclerosis (MS) reduce the levels of main ovarian reserve markers, measured through automated systems? Summary answer There is no statistically significant difference in ovarian reserve markers between women affected by Multiple Sclerosis and healthy controls. What is known already Few studies evaluated the issue of ovarian reserve in women affected by MS, so far. The majority of them observed no differences in ovarian reserve markers between women affected by MS and healthy controls. However, interestingly, women with MS in case of a more active disease were observed to have lower levels of ovarian reserve markers. In fact, evidence suggests that an inflammatory status may reduce the ovarian reserve, where autoimmune-related diseases are considered a possible cause of Premature Ovarian Insufficiency (POI). Study design, size, duration This is a single-center multidisciplinary prospective case-control (44 cases and 72 controls) study performed from September 2019 to September 2022. Participants/materials, setting, methods Cases were reproductive-age women affected by MS recruited at our MS unit, controls were age- and body mass index (BMI) - matched healthy women recruited at our assisted reproductive technology (ART) unit. During the early follicular phase, a transvaginal ultrasound was performed with sonoAVC software (GE medical systems) for 3D antral follicle count and a blood sample taken for hormonal assays (Anti-Müllerian hormone levels was measured using the Roche Elecsys AMH Plus). Main results and the role of chance There are no differences in age (p = 0.11), BMI (p = 0.96) as per protocol. Moreover, differences in FSH (p = 0.14), LH (p = 0.13), estradiol (p = 0.92), and AMH (p = 0.99) levels were not statistically significant. The 3D antral follicle count showed a statistically significant difference (p = 0.03), but following a re-evaluation of pictures in post-processing, removing the small antral follicles (2-4 mm) the difference was not statistically significant (p = 0.14). Limitations, reasons for caution We could not assess women before they started on medication for MS. Therefore, we could not exclude that medication have an impact in these results, also because the sample is small and patients were not treated with the same drug. Moreover, all patients were in disease remission at enrollment. Wider implications of the findings This is the first study to evaluate ovarian reserve markers with automated systems. Our results confirmed the ones previously published. However, the interesting data on 3D antral follicle count increased in MS women should be confirmed and studied in order to evaluate if follicular dynamics are altered in Multiple Sclerosis. Trial registration number Not applicable

P-582 Identification of microRNA in women diagnosed with premature ovarian insufficiency using urinary exosome in Korean population

Abstract Study question Would we able to identify specific microRNA which is specific in premature ovarian insufficiency? Summary answer Our study provides compelling evidence suggesting that microRNA-4516 would be a diagnostic marker in Korean premature ovarian insufficiency women. What is known already Premature ovarian insufficiency is diagnosed in women before the age of 40 years old, when there is no menstruation for at least for six months with hypergonadotropic hypogonadism in serum evaluation. It is a disease which ovarian functions are irreversible, but women visit outpatient clinic only when their ovarian functions are already diminished. Exosomes are functional vehicles which transport complex proteins, lipids and nucleic acids between cells, containing numerous microRNAs and it may include disease-specific miRNA signature that can be a valuable diagnostic tool. Study design, size, duration This was a prospective study including patients diagnosed with premature ovarian insufficiency(POI)/Turner syndrome and control individuals who visited outpatient clinic in Department of Gynecology in Gil Hospital, Gachon University from January 2019 to August 2021. We divided the patients into two groups depending on the time period of enrollment; Sequencing cohort and Validation cohort. In sequencing cohort, total 19 patients (7/7/5) were enrolled, and in validation cohort, total 46 patients (15/11/20) were enrolled. Participants/materials, setting, methods The patients in POI group were participants diagnosed with POI but they not Turner syndrome. After collecting first morning urine samples, we extracted exosomes and RNA sequencing was done based on microRNA library. We did real-time PCR using a selected hsa-miR-4516 for confirmation. Then we did confirmation study using POI mouse model (Cyclophosphamide + Busulfan). Main results and the role of chance Expression of hsa-miR-4516 detected via qRT-PCR analysis of RNA from the validation cohort exactly mapped the RNA sequencing results of the sequencing cohort. Moreover, it was significantly upregulated in patients with POI and Turner syndrome compared to the control group. The TaqMan PCR assay confirmed the upregulation of hsa-miR-4516 expression in patients with POI and Turner syndrome. To evaluate miR-4516 as a biomarker of POI, we established a chemotherapy-induced POI mouse model by injecting the mice with CTX and BUS. Next, we assessed the expression of mmumiR-4516 in the ovaries of the chemotherapy-induced POI mouse model; mmu-miR-4516 expression was significantly increased in the ovaries but not in the uteri in the POI mouse model compared to the control. These results suggest that upregulation of mmumiR-4516 expression is associated with pathological changes in the ovaries. Limitations, reasons for caution We also observed inconsistencies between the sequencing and qRT-PCR results. The cause of POI is highly diverse, which can lead to discrepancies in exosomal miRNA expression between the cohorts. The cohort is small and limited compared to the validation cohort, and this may have caused inconsistencies between the cohorts. Wider implications of the findings We suggest that hsa-miR-4516 could be developed as a biomarker of POI within an easy and convenient diagnostic test using non-invasive sampling methods. Trial registration number -

Analysis of oxidative stress and antioxidative potential in premature ovarian insufficiency.

Premature ovarian insufficiency (POI) is characterized by an early decline in ovarian function, inducing secondary amenorrhea. While the cause of POI has not yet been identified, the function of mitochondria in the ovaries and the cytotoxicity associated with reactive oxygen species (ROS) have been implicated in follicle pool depletion and a decline in follicle quality. Recently developed tests have enabled easy measurement of diacron-reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP). The combination of these two tests is used to comprehensively assess oxidative stress in the blood. To comprehensively assess the oxidative stress of d-ROMs and BAP in POI. Participants were classified into two groups: A POI group of 11 women aged < 40 years examined between January 2021 and June 2022 with a history of secondary amenorrhea for at least 4 mo in our hospital and an FSH value of ≥ 40 mIU/mL; and a control group of healthy women of the same age with normal ovarian function in our hospital. Plasma d-ROMs and BAP were measured in both these groups underwent. Differences between groups were assessed using the t-test. The mean age and mean body mass index (BMI) were 35.8 ± 3.0 years and 20.1 ± 1.9 kg/m2 in the control group and 35.8 ± 2.7 years and 19.4 ± 2.5 kg/m2 in the POI group, respectively. The mean gravidity and parity in control and POI groups were 0.6 ± 0.7 and 0.4 ± 0.5 and 0.6 ± 0.9 and 0.3 ± 0.5, respectively. The two groups did not differ significantly in terms of mean age, BMI, gravidity, or parity. The d-ROMs level was significantly higher in the POI group than in the control group (478.2 ± 58.7 vs 341.1 ± 35.1 U.CARR; P < 0.001); however, the BAP level did not significantly differ between the two groups (2078.5 ± 157.4 vs 2029.0 ± 186.4 μmol/L). The oxidase stress index (d-ROMs/BAP × 100) was significantly higher in the POI group than in the control group (23.7 ± 3.3 vs 16.5 ± 2.1; P < 0.001). Oxidative stress was significantly greater in the POI group than in the control group, suggesting oxidative stress as a factor that can serve as a POI biomarker.

Premature Ovarian Insufficiency - diagnostic and therapeutic challenges

Introduction and purpose: Premature Ovarian Insufficiency (POI) is a rare but serious disease diagnosed in about 1% of women before the age of 40. It is characterized by hypoestrogenism and an increase in the level of the follicle-stimulating hormone, which is associated with menstrual disorders, pregnancy failures and a reduced quality of life. The main causes of POI include a genetic background and autoimmune abnormalities, as well as increasingly jatrogenic factors. Care for patients is focused on hormone replacement therapy and infertility treatment. The aim of this review is to discuss current treatment methods, the impact of early diagnosis and the prevention of premature ovarian insufficiency, which affects the ability to have one's own offspring. The article also describes new methods of infertility treatment that are under investigation.&#x0D; Materials and methods: This literature review is based on articles published in the PubMed database from 2015 to 2022 using the following phrases: premature ovarian insufficiency, premature ovarian insufficiency stem cells&#x0D; State of knowledge: The only way to treat infertility in women with POI is egg donation. Currently, research is being conducted to achieve early diagnosis, introduce new treatment methods and reduce the number of iatrogenic forms of POI.&#x0D; Conclusion: The main problem for women diagnosed with POI is the difficulty or often impossibility of having their own children. Egg donation does not allow for the transfer of the patient's genetic material, and is also not available as a treatment option in some countries. For this reason, new methods for early diagnosis, treatment, and prevention of POI are being developed.

The hypergonadotropic hypogonadism conundrum of classic galactosemia.

Hypergonadotropic hypogonadism is a burdensome complication of classic galactosemia (CG), an inborn error of galactose metabolism that invariably affects female patients. Since its recognition in 1979, data have become available regarding the clinical spectrum, and the impact on fertility. Many women have been counseled for infertility and the majority never try to conceive, yet spontaneous pregnancies can occur. Onset and mechanism of damage have not been elucidated, yet new insights at the molecular level are becoming available that might greatly benefit our understanding. Fertility preservation options have expanded, and treatments to mitigate this complication either by directly rescuing the metabolic defect or by influencing the cascade of events are being explored. The aims are to review: the clinical picture and the need to revisit the counseling paradigm; insights into the onset and mechanism of damage at the molecular level; and current treatments to mitigate ovarian damage. In addition to the work on this topic by the authors, the PubMed database has been used to search for peer-reviewed articles and reviews using the following terms: 'classic galactosemia', 'gonadal damage', 'primary ovarian insufficiency', 'fertility', 'animal models' and 'fertility preservation' in combination with other keywords related to the subject area. All relevant publications until August 2022 have been critically evaluated and reviewed. A diagnosis of premature ovarian insufficiency (POI) results in a significant psychological burden with a high incidence of depression and anxiety that urges adequate counseling at an early stage, appropriate treatment and timely discussion of fertility preservation options. The cause of POI in CG is unknown, but evidence exists of dysregulation in pathways crucial for folliculogenesis such as phosphatidylinositol 3-kinase/protein kinase B, inositol pathway, mitogen-activated protein kinase, insulin-like growth factor-1 and transforming growth factor-beta signaling. Recent findings from the GalT gene-trapped (GalTKO) mouse model suggest that early molecular changes in 1-month-old ovaries elicit an accelerated growth activation and burnout of primordial follicles, resembling the progressive ovarian failure seen in patients. Although data on safety and efficacy outcomes are still limited, ovarian tissue cryopreservation can be a fertility preservation option. Treatments to overcome the genetic defect, for example nucleic acid therapy such as mRNA or gene therapy, or that influence the cascade of events are being explored at the (pre-)clinical level. Elucidation of the molecular pathways underlying POI of any origin can greatly advance our insight into the pathogenesis and open new treatment avenues. Alterations in these molecular pathways might serve as markers of disease progression and efficiency of new treatment options.

Potential therapeutic drug targets and pathways prediction for premature ovarian insufficiency —Based on network pharmacologic method

Ethnopharmacological relevanceThe incidence of premature ovarian insufficiency (POI) is gradually increasing, the proportion is rising especially in female infertility patients. The risk of death of POI patients with cardiovascular disease also increases significantly. The cause of POI is complex and unclear, and clinical treatment is still in the exploratory stage, are two major constraints of treating POI. Traditional Chinese medicine (TCM) is widely used in the treatment of POI, and it is a good way to combine the development of modern new drugs with the help of TCM to predict the therapeutic targets. Aim of the studyIn this study, four herbs commonly used in clinical treatment of POI, namely Radix Paeoniae, Polygonatum sibiricum, Rehmannia glutinosa and Eucommia ulmoides were selected to predict their mechanism in the treatment of POI, using network pharmacology methods. Then verify the predicted targets by animal test. Aim to find more effective POI potential core treatment targets and main pathways. Materials and methodsWe screened the active ingredients of drugs from the TCM System Pharmacology Analysis Platform (TCMSP), Performed target prediction of active ingredients from databases such as SwissTargetPrediction and compare and analyze the POI-related targets retrieved from them to obtain potential targets for drug treatment of POI. Used STRING database to construct a protein interaction network, Cytoscape 3.7.2 software to construct an active ingredient-target-pathway network, and DAVID database to conduct the Kyoto Encyclopedia of Genes and Genomes (KEGG) on the intersection targets and gene ontology (GO) enrichment analysis. ResultsThe result is: there were 25 key targets for the treatment of POI with Radix Paeoniae Alba, 31 for the treatment of POI by Eucommia ulmoides, 28 for the treatment of POI by Polygonatum sibiricum, and 8 key targets for the treatment of Rehmannia glutinosa. The intersection targets of four herbs were defined as the core targets, which are CYP19A1, EGF, ESR1, ESR2, MDM2, AR, PCYP17A1, PPARG. Four Chinese herbs treat POI mainly through HIF-1 signaling pathway, PI3K-Akt signaling pathway, FoxO signaling pathway, Estrogen signaling pathway etc. A mouse model of POI was constructed based on the results of network pharmacology to verify the predicted targets. The results showed that the protein expression of the core target changed, and the estrogen level was increased by reducing the expression of peroxisome proliferator-activated receptor gamma (PPARG). ConclusionsThis study predicts the mechanism of multiple herbs in the treatment of POI, screens out more potential therapeutic drug targets and main pathways of POI treatment and provides new ideas for the subsequent development of POI therapeutic drugs.

The effects and mechanism of taxanes on chemotherapy-associated ovarian damage: A review of current evidence.

Chemotherapy is often a cause of premature ovarian insufficiency and infertility since the ovarian follicles are extremely sensitive to the effects of chemotherapeutic agents. Different chemotherapeutic agents with varying mechanisms of action may damage ovarian function differently. Taxanes are widely used in clinical cancer treatment, but the specific reproductive toxicological information is still controversial. This review described the impact and duration of taxanes on ovarian function in women and analyzed the possible reasons for different conclusions. Furthermore, the toxicity of taxanes on ovarian function and its possible mechanisms were discussed. The potential protective strategies and agents against ovarian damage induced by taxanes are also reviewed.

Stem Cell-Based Therapeutic Strategies for Premature Ovarian Insufficiency and Infertility: A Focus on Aging

Reproductive aging is on the rise globally and inseparable from the entire aging process. An extreme form of reproductive aging is premature ovarian insufficiency (POI), which to date has mostly been of idiopathic etiology, thus hampering further clinical applications and associated with enormous socioeconomic and personal costs. In the field of reproduction, the important functional role of inflammation-induced ovarian deterioration and therapeutic strategies to prevent ovarian aging and increase its function are current research hotspots. This review discusses the general pathophysiology and relative causes of POI and comprehensively describes the association between the aging features of POI and infertility. Next, various preclinical studies of stem cell therapies with potential for POI treatment and their molecular mechanisms are described, with particular emphasis on the use of human induced pluripotent stem cell (hiPSC) technology in the current scenario. Finally, the progress made in the development of hiPSC technology as a POI research tool for engineering more mature and functional organoids suitable as an alternative therapy to restore infertility provides new insights into therapeutic vulnerability, and perspectives on this exciting research on stem cells and the derived exosomes towards more effective POI diagnosis and treatment are also discussed.

Erken Yumurtalık Yetersizliğinde Kadın İnfertilitesinin Optimizasyonu

Premature ovarian failure is a complex pathology with numerous etiologies and multiple system sequelae resulting for early deprivation of ovarian sex hormones. In the studies to determine the etiology, it is seen that many patients are in the unexplained group. On the other hand, genetic predisposition and autoimmune cause of premature ovarian insufficiency (POI) are the most common known etiologic causes. Early diagnosis and energetic treatment are important in order to prevent symptoms related to estrogen deficiency. Initiating hormone replacement therapy as soon as possible will prevent menopausal symptoms and reduce long-term complications in women. Another important problem in these patients is infertility, which occurs due to ovarian failure. In fact, fertility, which begins to decrease before menstrual irregularity, turns into infertility with a rapid decline in the number of follicles in the following period. Here, the detection of patients at risk and appropriately timed fertility preservation treatments (such as freezing of oocyte, embryo, or ovarian tissue) or assisted reproductive techniques can be offered. Besides this, studies like stem cell therapy, platelet-rich plasma (PRP), and in vitro activation of ovarian tissue in patients with POI are recent and still investigational but may be promising in the future. In the present review, the current pathophysiology and treatment options of premature ovarian failure were discussed.

Rapid ovarian transcript changes during the onset of premature ovarian insufficiency.

Premature ovarian insufficiency (POI) affects 1-3% of women under 40 years of age. The identified causes are highly heterogeneous, and 70% of the cases are idiopathic. The ovarian manifestation varies from a variable population of follicles that fail to develop (follicular POI) to the absence of follicles (afollicular POI) with a transition from one to the other over time. Previously, we have described a mouse model of POI that results from an oocyte-specific deletion of N- and O-glycans; Double Mutant (DM). DM females produce only one litter before undergoing POI due to ovarian dysfunction. In this study, we have characterised the gene expression profile of prepuberal (3 weeks), fertile (6 weeks) and infertile (9 weeks) DM ovaries. Up-regulation of cathepsin K (Ctsk, with unknown ovarian function) seems to trigger transcriptional changes in DM ovaries. Significant transcriptional changes then occur rapidly, associated with morphophysiological changes displayed by DM mice throughout the onset of POI. We identified genetic pathways such as extracellular matrix and immune response as candidates for the onset of POI in DM females. Remarkably, DM mice and POI women share a set of differentially expressed genes, including a functionally and co-expressed network of Mcm (minichromosome maintenance proteins) family members. The transcriptomic profile of the DM mouse model provides novel insight into the aetiology of POI.Lay summaryProblems in ovary function lead to reduced fertility or infertility. One such condition is premature ovarian insufficiency (POI) which affects 1% of women under 40 years of age, and in over 70% of these, the cause of POI is unknown. To investigate POI, we have developed a mouse model. These mice are initially fertile but develop POI by 3 months of age. In this study, we investigated the changes in genes activated in the ovaries during the transition from fertility to POI, and we did this by comparing them to normal mice; gene activation leads to molecule production. A molecule known as cathepsin K seems to trigger changes during the onset of POI, followed by others related to structure and immune response pathways. In addition, some genes were identified that are similar between the POI mice and POI women.

O-147 Genomic analyses in 101,127 UK women show that previously reported monogenic genes are not common causes of premature ovarian insufficiency

Abstract Study question What is the penetrance of variants in previously reported genes for premature ovarian insufficiency (POI) in the general population? Summary answer Heterozygous genetic variants in previously reported monogenic genes are not a common, highly penetrant cause of POI. What is known already Premature ovarian insufficiency (POI), defined as menopause before 40, has been associated with variants in over 70 genes. However, the evidence to support the causality of individual genetic variants varies between studies. As genomic data becomes more accessible, it is essential to ascertain the penetrance of variants in the absence of family history information. We identified 75 genes associated with POI from the literature, including family segregation studies, consanguineous pedigrees and cohort analyses of whole-exome/targeted sequencing data. Functional evaluation was also available for many of the genes. Study design, size, duration We used data from 101,127 females of European ancestry in the UK Biobank, to study the role of previously reported monogenic causal genes on ovarian function. We tested the association of 301 previously reported variants with POI and ANM, plus more than 2.5 million rare variants which were annotated and had not been implicated in POI previously. Genomic variants were tested individually and also combined into a gene burden test. Participants/materials, setting, methods Age at natural menopause (ANM) was derived from self-reported questionnaire data from the age at last menstrual period, excluding those with surgical menopause or taking hormone replacement therapy. POI cases were classified as women with ANM under 40 years (N = 2,213). Linked primary care records were used to identify a clinical cohort of POI cases, including primary amenorrhea (N = 113). Main results and the role of chance All of the previously reported individual variants we identified in our POI cases were also detected in the control group. Moreover, the gene burden tests were not associated with POI or ANM as a quantitative trait. Our results indicate that autosomal dominant causes of POI are rare; a single predicted loss-of-function (LOF) or non-synonymous genetic variant in one of the previously reported genes is generally not pathogenic. Limitations, reasons for caution The penetrance of monogenic disease-causing variants is likely to be lower in population-based cohorts such as UK Biobank than in clinically-ascertained cohorts. Wider implications of the findings Heterozygous LOF or non-synonymous variants in previously reported POI genes should be interpreted with caution and are unlikely to cause POI. The findings have implications for clinicians diagnosing causes of POI. Trial registration number Not applicable

O-265 Luteinizing hormone is able to protect reproductive health in cancer patients

Abstract Study question Is luteinizing hormone (LH) able to protect ovarian follicles against cyclophosphamide (CPM)-induced damage in women? Summary answer LH significantly reduces primordial follicle (PMF) loss in ovarian cortical strips cultured in vitro with phosphoramide mustard (PM), the active metabolite of CPM. What is known already Cancer therapies are cause of premature ovarian insufficiency (POI) in female patients, since they induce a severe reduction of PMFs. For this reason, several compounds have been analyzed as adjuvant therapies to protect the ovarian reserve without interfering with cancer treatment on tumor cells. We recently demonstrated the protective effect of LH against ovotoxicity induced by PM and cisplatin on the ovary of prepuberal and adult mice. These results suggest the possibility to use LH prior or in concomitance with anticancer drugs in order to preserve oocytes in human patients, therefore preventing the early onset of menopause and/or infertility. Study design, size, duration Ovarian cortical tissues were collected from nine patients (age ± SD: 15.33 ± 4.50) who have cryopreserved their tissue before receiving anticancer treatment. For each patient, ovarian cortical strips were thawed and randomly assigned to the experimental conditions: Control (CTRL), PM and PM+LH. LH was added 1 hour before the treatment with PM. Samples were analyzed after 8, 16, 24 and 48 hrs of treatment. Participants/materials, setting, methods Ovarian cortial strips were placed onto culture plate insert and cultured for a maximum of 48 hours, in αMEM with 10% Serum Substitute Supplement with/out 200mIU/ml LH and/or 10μM PM. The samples were processed for: - Histology, for PMFs and primary follicles (PFs) analysis; - Immunohistochemistry, for the expression of protein involved in DNA damage, apoptosis and follicle activation; - Real-Time PCR, for the expression of apoptotic and inflammation related genes. Main results and the role of chance The follicle density in the untreated group varied from 233.03 to 3420.27 PMFs/mm3 between the nine patients analysed. Relative follicular density (%) was performed to analyse statistical differences between the groups. Relative PMFs density (%) was significantly reduced in PM vs CTRL either after 24 and 48 hrs, while this reduction was significantly counteracted by LH (24 hrs: CTRL=76.19±2.12; PM = 44.95±5.06; PM+LH=73.97±11.64. 48 hrs: CTRL=66.47±3.96; PM = 36.76±3.96; PM+LH=55.74±8.72). To investigate the mechanism underlying the observed effects, the expression of markers involved in DNA damage (gH2AX), apoptosis (Cleaved caspase 3, NOXA, PUMA), follicle activation (p-AKT, FOXO3a), cell cycle arrest (p21, Ki67), and inflammation (IL1β, TNFα) were analysed. The results showed that LH did not prevent DNA damage induced by PM, since gH2AX positive oocytes were seen both in PM and PH+LH group at 16h; at 24hrs however we observed a downregulation of the gH2AX expression in the PH+LH group (PM@ 100% vs PM+LH@35%). LH also counteracts the activation of chemotherapy-induced apoptotic processes, by reducing the levels of pro-apoptotic factors such as NOXA, PUMA and CC3, and follicles activation lowering AKT-FOXO3a signaling axis. Moreover, PM treatment creates a proinflammatory microenvironment, as shown by increased IL1β and TNFα gene expression, partially counteracted by LH pretreatment. Limitations, reasons for caution Experiments are carried out in vitro; the functionality of the cortex in xenografts should be evaluated. Careful evaluation of which patients might use this protector is needed. Wider implications of the findings These findings demonstrate that LH is able to reduce PMFs loss in human ovarian cortex exposed to PM. These results encourage thinking about the use of the hormone as a ferto-protector agent in clinical trials to prevent the premature onset of menopause and/or infertility in women undergoing anticancer treatments. Trial registration number not applicable

104 Premature ovarian insufficiency: Should genetic testing be tailored?

Objective: Premature Ovarian Insufficiency (POI) is described as amenorrhea due to loss of ovarian function before the age of 40 and is characterized by elevated gonadotropins. Possible causes of POI are chromosomal, genetic, infectious, autoimmune and iatrogenic. However, the majority of cases are “idiopathic”.[ [1] European Society for Human Reproduction and Embryology (ESHRE) Guideline Group on POI; Webber L, Davies M, Anderson R, et al. ESHRE Guideline: management of women with premature ovarian insufficiency. Hum Reprod 2016;31(5):926–37. Google Scholar ] Pedigrees of families with non-syndromic POI have shown an inheritance pattern of dominant or recessive transmission, autosomal or X-linked, with variable expressivity and penetrance. Moreover, several syndromes attributed to specific genetic mutations may include POI in their clinical manifestations.[ [2] Qin Y. Jiao X. Simpson J.L. Chen Z.-J. Genetics of primary ovarian insufficiency: new developments and opportunities. Hum Reprod Update. 2015; 21: 787-808 Crossref PubMed Scopus (265) Google Scholar ] The aim of this study was to identify women whose family or personal history would be more likely to indicate a genetic mutation as a cause of POI.

Premature ovarian insufficiency.

The natural lifespan of the ovary is occasionally interrupted by pathological processes; some are known, but many are unknown. Premature ovarian insufficiency (POI) can be a devastating diagnosis for an adolescent or for someone who has yet to start a family. Common causes of POI include genetic and chromosomal defects, autoimmune damage, and cancer treatments. Knowledge of the pathogenesis of this condition and an awareness of contemporary hormone replacement and fertility options are required to design a multidisciplinary therapeutic approach comprising reproductive medicine, endocrinology, clinical psychology, and assisted fertility expertise.

New STAG3 gene variant as a cause of premature ovarian insufficiency

RESUMENObjetivos: describir un caso de falla ovárica secundaria a una variante patogénica homocigota en el gen STAG3 no reportada previamente.Materiales y métodos: paciente de 16 años con amenorrea primaria y ausencia de características sexuales secundarias, en quien se documentó hipotiroidismo autoinmune, pobre desarrollo genital y cintilla gonadal, por lo cual se realizó secuenciación de exorna clínico. Se identificó una variante homocigota patogénica previamente no reportada en el gen STAG3, el cual ha sido relacionado con insuficiencia ovárica prematura (IOP).Conclusiones: en este caso, la realización de exorna clínico fue determinante para identificar una alteración del gen STAG, probablemente asociada a la IOP y el pronóstico a largo plazo de la paciente. Se establece una nueva variante patogénica c.2773delT; p.Ser925Profs*6 del gen STAG3 asociada a la IOP.