Low Functional Ovarian Reserve Research Articles

Extended LH administration as a strategy to increase the pool of recruitable antral follicles in hypothalamic amenorrhea: evidence from a case series.

New evidence is indicating a growing role of LH in promoting ovarian follicular growth and maturation, even at the early stages. LH seems to enhance the transition of follicles to the antral stage hence leading to an increase in the so-called functional ovarian reserve (recruitable antral follicles). Hypogonadotropic hypogonadism is characterized by low, and sometimes undetectable, serum LH and FSH levels, which may indeed explain the low anti-Müllerian hormone (AMH) levels and antral follicle count (AFC) found in patients affected by this condition. We report here the cases of two young women affected by hypothalamic amenorrhea (HA) that presented for fertility treatment with very low functional ovarian reserve. The two patients were treated with exogenous LH for 1 and 2 months (extended LH administration: ELHA) at the dose of 187.5 IU LH every day and 150 IU LH every other day, respectively. In both the cases there was an increase in serum AMH levels and in the AFC. In one patient, the AMH and AFC increased from a baseline 1.3 ng/ml and 8 to 2.3 ng/ml and 14 at end of treatment, respectively. In the second case, serum AMH and AFC increased from 0.4 ng/ml and 6 to 1.6 ng/ml and 13, respectively. One patient underwent ovarian stimulation before and after ELHA, showing an increase in the number of mature oocytes recruited (3 versus 8 metaphase II (MII) oocytes before and after, respectively). The second patient underwent an IVF cycle after ELHA resulting in the retrieval of six MII oocytes and an ongoing pregnancy following transfer of a single blastocyst. Women with HA are characterized by chronic, low levels of gonadotrophins, which may impact not only on the cyclic recruitment of follicles but also the progression of small growing follicles through the first stages of folliculogenesis. Some women with HA may in fact show very low serum AMH and AFC. Our case series shows that the administration of LH at a dose of at least 150-187.5 IU every day or every other day may contribute to a clinically evident increase in the functional ovarian reserve (AFC), and probably accounts for a positive effect of LH on the progression of follicles throughout the early stages of folliculogenesis.

P-645 Intra-ovarian injection of autologous platelet-rich plasma (PRP) in poor prognosis IVF patients within 1 year of treatment: a cohort study

Abstract Study question Does intraovarian injection of platelet-rich plasma (PRP) change parameters of ovarian function in patients with evidence of diminished ovarian reserve? Summary answer Over one year of follow-up there was no evidence of a positive effect of PRP treatment on ovarian function. What is known already Several authors have previously reported positive outcomes and improved response in ovarian function after treatment with PRP. However, thus far, no randomized controlled trials and few cohort studies have been published. Study design, size, duration This is a longitudinal cohort study following ovarian response among 80 patients with low functional ovarian reserve (LFOR) defined by AMH < 1.1 ng/mL, FSH > 12 mIU/mL or at least one prior IVF cycle with ≤ 3 oocytes for one year before and one year after intraovarian treatment with PRP. Participants/materials, setting, methods 1.5 cc of PRP was injected into the cortex of ovaries in an average 12 injections per ovary. Beginning one month after their PRP treatment, patients underwent one or more cycles of ovulation induction for IVF. Outcome measures were endocrine response, and numbers of oocytes and embryos produced in by patients in response to a maximal gonadotropin stimulation before and after PRP treatment. Main results and the role of chance In this study women between ages 28 and 54 failed to demonstrate statistically significant outcome benefits from intra-ovarian PRP. Two very poor-prognosis 40-year-old patients with prior failed IVF cycles at other IVF centers that never reached embryo transfer, however, achieved pregnancy in an IVF cycle following PRP treatment for an ongoing pregnancy rate of 4.7% if a patient following PRP achieved retrieval of at least 1 oocyte with IVF. Limitations, reasons for caution This is an observational study of patients who performed poorly in past ovulation induction cycles, thus any improvement observed could have been accounted for by regression to the mean. The occurrence of two pregnancies in this group could be secondary to eventual success following a string of previous failures. Wider implications of the findings Even extremely poor prognosis will occasionally achieve a successful outcome from IVF. Uncontrolled case series cannot provide sufficient evidence to support routine use of an intervention like PRP. Future studies demand the use of well- controlled randomized trials. Trial registration number NCT04275700

Preliminary report of intraovarian injections of autologous platelet-rich plasma (PRP) in extremely poor prognosis patients with only oocyte donation as alternative: a prospective cohort study.

STUDY QUESTIONDoes intraovarian injection of platelet-rich plasma (PRP) change ovarian function in patients with extremely low functional ovarian reserve (LFOR) who, otherwise, would likely only have a chance of pregnancy through third-party oocyte donation?SUMMARY ANSWERNo clinically significant effects of PRP treatment on ovarian function were observed over 1 year of follow-up.WHAT IS KNOWN ALREADYSeveral investigators have reported improved responses to ovulation induction after treatment with PRP. However, previous published reports have involved, at most, only small case series. Whether PRP actually improves ovarian performance is, therefore, still unknown. PRP is nevertheless widely offered as an ‘established’ fertility treatment, often under the term ‘ovarian rejuvenation’.STUDY DESIGN, SIZE, DURATIONWe are reporting a prospective cohort study of 80 consecutive patients at ages 28–54 with LFOR, defined by anti-Müllerian hormone <1.1 ng/ml, FSH >12 mIU/ml or at least one prior IVF cycle with ≤3 oocytes within 1 year. The women were followed for 1 year after an intraovarian PRP procedure.PARTICIPANTS/MATERIALS, SETTING, METHODSPRP (1.5 ml) was injected into the cortex of ovaries with an average of 12 injections per ovary. Study participants were followed every 3 days for 2 weeks after PRP treatment with estradiol and FSH measurements and vaginal ultrasound to observe follicle growth and thereafter followed weekly. Beginning 1 month after their PRP treatment, participants underwent one or more cycles of ovarian stimulation for IVF. Outcome measures were endocrine response, and numbers of oocytes and embryos produced in response to a maximal gonadotropin stimulation before and after PRP treatment.MAIN RESULTS AND THE ROLE OF CHANCEIn this study, women failed to demonstrate statistically significant outcome benefits from intraovarian PRP. However, two 40-year-old very poor-prognosis patients, with prior failed IVF cycles that never reached embryo transfer at other centers, achieved pregnancy, resulting in an ongoing pregnancy rate of 4.7% among patients who, following PRP, produced at least one oocyte (n = 42).LIMITATIONS, REASONS FOR CAUTIONAs an observational study of patients who performed poorly in past ovarian stimulation cycles, the improvement may be accounted for by regression to the mean. Similar considerations may also explain the occurrence of the two pregnancies.WIDER IMPLICATIONS OF THE FINDINGSThis study demonstrates that, even in extremely poor prognosis patients due to LFOR, sporadic pregnancies are possible. The study, however, does not allow for the conclusion that those pregnancies were the consequence of PRP treatments. A case series, indeed, does not allow for such conclusions, even if results are more suggestive than here. This registered study, therefore, must be viewed as a preliminary report, with further data expected from this study but also from two other prospectively randomized ongoing registered studies with more controlled patient selection.STUDY FUNDING/COMPETING INTEREST(S)This work was supported by intramural funds from The Center for Human Reproduction and the not-for-profit research Foundation for Reproductive Medicine, both in New York, NY, USA. N.G. and D.H.B. are listed as co-inventors on several US patents. Some of these patents relate to pre-supplementation of hypo-androgenic infertile women with androgens, such as dehydroepiandrosterone and testosterone and, therefore, at least peripherally relate to the subject of this manuscript. They, as well as D.F.A., have also received research support, travel funds and speaker honoraria from several pharmaceutical and medical device companies, though none related to the here presented subject and manuscript. N.G. is a shareholder in Fertility Nutraceuticals and he and D.H.B. receive royalty payments from Fertility Nutraceuticals LLC. E.M. has no conflicts of interest to declare.TRIAL REGISTRATION NUMBERNCT04275700

Investigating the Clinical Utility of the Anti-Mullerian Hormone Testing for the Prediction of Age at Menopause and Assessment of Functional Ovarian Reserve: A Practical Approach and Recent Updates.

Low ovarian reserve is a serious condition, leading to sterility in up to 10% of women in their mid-thirties. According to current knowledge, serum anti-Müllerian hormone (AMH) levels for age are the best available marker for the screening the quantity of a woman’s functional ovarian reserve, better than age alone or other reproductive markers. This review summarizes recent findings, clinical utility and limitations in the application of serum AMH testing as an accurate marker for the screening of functional ovarian reserves and predicting age at menopause. AMH assessment hold promise in helping women make informed decisions about their future fertility and desired family size. However, screening of the functional ovarian reserve could be offered to all women at 26 years of age or older who seek to assess future fertility or in case of personal request, ovarian reserve screening may be considered beyond 30 years; however, it has never been advocated beyond 35 years, since it is not advisable to delay childbearing beyond this age. In this respect, an age-specific serum AMH levels lower than the 10th percentile may be used as a threshold for the identification of a low functional ovarian reserve in an individual woman. Its level should be interpreted with caution in the adolescent and young women aged below 25 years (since AMH levels peak at this age); recent users of hormonal contraceptives (since AMH levels transiently decrease until two months after discontinuation); and women with PCOS (which dramatically increases AMH levels). However, the ability of AMH levels to predict the time to menopause is promising but requires further investigation and routine AMH testing for the purposes of predicting the time to menopause is not recommended.

A PILOT STUDY OF OOCYTE PRODUCTION AND ENDOCRINE RESPONSE AFTER INTRAOVARIAN TREATMENT WITH PLATELET-RICH PLASMA (PRP) IN INFERTILE WOMEN OF VERY ADVANCED AGE

Though proper validation studies are lacking, intraovarian PRP is widely being promoted as treatment to improves ovarian response in women with low functional ovarian reserve (LFOR). The objective of this prospective pilot study was, therefore, to compare in women of very advanced ages ovarian responses before and after a PRP procedure. This study involved 2 distinct groups of patients, both characterized by very advanced age: Group 1 were women with LFOR but still regular menstrual cycles; Group 2 were LFOR patients but with irregular menstrual cycles or amenorrhea. All patients in both groups had undergone prior failed IVF cycles. Before receiving a PRP treatment all patients received at least 6 weeks of pre-supplementation with DHEA (25mg TID) and CoQ10 (1000mg/day). The patient’s PRP was extracted from their own blood as previously reported,1 and was in early follicular phase under ultrasound control injected into ovaries subcapsular in 5-10 needle insertions per ovary, with the patient under subconscious sedation. IVF cycles were initiated on day-2 of subsequent menstrual period with 450 IU of FSH and 150IU of hMG (different manufacturers). Primary study outcomes were AMH levels and oocyte numbers retrieved. This still ongoing study is registered at ClinicalTrials.gov as NCT04275700. In Group 1 (n=28) average age was 45.3 ± 4.6 years. AMH pre-PRP 0.36 ± 0.6, post- PRP 0.45 ± 0.5 N.S.); Oocytes pre-PRP 1.99 ± 1.5, post-PRP 2.12 ± 1.4 (N.S.). Group 2 (n=9) average age was 45.3 ± 6.1 years. AMH pre-PRP 0.13 ± 0.29 post- PRP 0.23 ± 0.4; (N.S.); Oocytes pre-PRP 1.62 ± 2.1, post-PRP 1.3 ± 1.2 (N.S). In here investigated 2 groups of patients PRP administration does not appear to have significantly affected ovarian reserve, as documented by unchanged AMH levels and similar oocyte yields before and after PRP.

P–600 Ovarian function following intraovarian injection of autologous platelet rich plasma (APRP) in women with low functional ovarian reserve

Abstract Study question Does exposure of ovaries to autologous growth factors in platelet rich plasma (APRP) affect the pituitary ovarian axis? Summary answer Within 60 days after injection, growing follicle numbers and estrogen levels increased, though FSH did not change, with effects most pronounced in still menstruating women. What is known already APRP is extracted from a patient’s autologous blood and delivers growth factors. It is widely used in several medical specialties and has in infertility practice been reported to increase folicle/egg numbers if injected into ovaries and improve endometrial thickness/implantation if used for perfusion of the endometrium. Study design, size, duration Prospective observational cohort study of women with low functional ovarian reserve, followed for 60 days after subcortical injection of ovaries. Participants/materials, setting, methods 44 women with prior poor response to ovulation induction, FSH &amp;gt; 12 mIU/mL and AMH &amp;lt; 1.0 ng/mL. APRP was prepared using Regen Lab PRP Kit which is approved by the US-FDA. 1.0–1.5 ml of PRP was injected into the cortex of each ovary divided among 7 to 10 injection sites. Participants were followed every three days with monitoring for estradiol, FSH and follicle growth for the first two weeks after PRP and then weekly. Main results and the role of chance 21/43 patients still regularly menstruated (subgroup A, age 43.9 ± 5.1 years); 23/43 (subgroup B, age 42.6 ± 6.2 years) were amenorrheic for a median of 6 months. In A, AMH, FSH and estradiol were 0.18 ± 0.20 ng/mL, 37.5 ± 47.6 mIU/mL, and 100.2 ± 73.4 pg/mL, while in B they were 0.06 ± 0.11 ng/mL, 73.0 ± 44.8 mIU/mL and 66.7 ± 57.6 pg/mL. Following APRP, A-patients demonstrated increased estradiol to 211 ± 193.7 pg/mL (P = 0.029) while B-patients only demonstrated a trend to 98.1 ± 86.5 (P = 0.09). Among A patients, 14/21 (66.7%) entered IVF cycles and 5/21 (23.8%) reached retrieval. So-far 1 patients established an ongoing clinical pregnancy. Among B patients 8/23 (34.8%) entered IVF cycles and only 2/23 (8.7%) reached retrieval and none achieved pregnancy. Limitations, reasons for caution This observational study was only carried out to estimate possible effects of APRP treatments. Based on these observations, we are now conducting a randomized controlled trial, limited to cycling women under age 45 years [registration # NCT04278313]. Wider implications of the findings: PRP appears to have limited ability to affect ovarian reserve of older, and especially amenorrheic women. It may, however, exert more favorable effects on still menstruating women. Promotion of APRP treatment as “ovarian rejuvenation,” however, appears to be an inappropriate choice of words. Trial registration number N/A

MON-354 Undiagnosed Addison's Disease as a Cause of Infertility

Background: There are multiple potential mechanisms for infertility in AD (Addison’s disease) including suppression of HPG axis, LFOR (low functional ovarian reserve) secondary to AH (adrenal hypoandrogenism) or concomitant autoimmune POF (premature ovarian failure), as well as suppressive effect from high 17-OHP (17α-hydroxyprogesterone). Nonetheless, there is scarce literature concerning this complication of AD. Clinical Case: A 28 year old female with a normal BMI and a history of hypothyroidism, suspected PCOS, and spontaneous abortion presented to RE (Reproductive Endocrinology) for a 2 year history of primary infertility. She had menarche at age 12, with regular menses for 6 years until OCP was initiated. At age 24 she spontaneously conceived, but unfortunately miscarried. She subsequently developed oligomenorrhea, reporting menses every 3-4 months in the absence of hirsutism or acne. Initial evaluation demonstrated negative β-hCG, normal TSH 1.27 mcIU/ml (0.27-4.2), high anti-TPO antibodies 1040 IU/ml (<34), progesterone 12.79 ng/ml (0.06-0.89 follicular, 1.83-23.9 luteal), AMH 13.5 ng/ml (1.03-11.1), and 17-OHP 272 ng/dl (<80 follicular, <285 luteal), and low DHEA-S 20.6 mcg/dl (98.8-340) and free testosterone 1.6 pg/ml (0.16.4). AFC (antral follicle count) was 50, and semen analysis and HSG were unrevealing. Elevated 17-OHP was thought to represent luteal phase, but repeat in early follicular phase remained elevated, prompting referral to Endocrinology. Co-syntropin stimulation test was performed with pre-ACTH 17-OHP persistently elevated (546 ng/dl), with minimal increase 1 hour post-ACTH (598 ng/dl). Cortisol failed to stimulate increasing only from 11 to 14 mcg/dl post-ACTH. Further evaluation revealed elevated ACTH (324 pg/ml), and positive anti-adrenal antibodies. Following diagnosis of AD, HC (hydrocortisone) 10 mg in the morning and 2.5 mg in the afternoon was initiated with improvement in menstrual regularity to 34 day cycles. RE recommended initiation of letrozole for ovulation induction, and intrauterine insemination. However, given improvement in cycles with HC, Endocrinology recommended delaying fertility treatments temporarily. Four months after HC initiation, pregnancy was confirmed, and she subsequently delivered a healthy baby girl. Conclusion: This case emphasizes the importance of a thorough endocrinologic evaluation in patients with infertility, particularly those with other autoimmune endocrinopathies. The etiology of infertility in AD is likely multifactorial, and there is minimal supportive literature to date. While some authors have proposed LFOR secondary to adrenal hypoandrogenism or POF as an underlying etiology, this unique case describes a patient with high AMH and AFC, who had successful conception and pregnancy following initiation of HC replacement.

The Impact of In vitro Oocyte Maturity on Developmental Potential of Embryos Derived from Controlled Ovarian Stimulation Cycles

Background and Aims: One of the major subjects for improving in vitro fertilization (IVF) outcome is the quantity and quality of retrieved oocytes. In vitro maturation (IVM) provides an opportunity for using immature oocytes routinely discarded in clinics. This study aimed at evaluating the quality of embryos derived from in vivo and rescue in vitro matured oocytes. Materials and Methods: Totally, 462 immature oocytes as cases and 466 mature (MII) oocytes as controls were included for study of their developmental competence. Oocytes underwent intracytoplasmic sperm injection insemination and then denuded oocytes were microscopically assessed regarding cytoplasmic and nuclear maturity and quality. Results: The morphological assessments showed fertilization rate of 60.9 and 61.4%, the embryo formation rate of 86.7% and 90.9% and arresting rate of 27.3% and 25.6% for the case and control oocytes, respectively. Evaluating embryo quality in the cleavage stage indicated that 63% of the embryos in the case group and 68% of the embryos in the control group were of good quality. There was no significant difference between fertility rate and arresting rate of oocytes matured in both groups, although the embryo formation rate and the quality of embryos differed significantly. Conclusions: Our findings suggest that IVM is a valuable and practical option for patients who had to cancel IVF treatment cycles because of severe responses or resistance to routine hormonal therapies or those with low functional ovarian reserve.

Developmental and cytogenetic assessments of preimplantation embryos derived from in-vivo or in-vitro matured human oocytes

Aneuploidy is of great relevance to embryo selection, as it represents one of the important causes of implantation failure. Furthermore, immature oocytes, retrieved during gonadotrophin-stimulated IVF cycles, are generally discarded in clinics; whereas, there was no detectable comprehensive evidence on higher rates of aneuploidy based on maturity status on the day of oocyte retrieval. As well, the correlation between embryo morphology on aneuploidy remains unclear. The aim was to evaluate the developmental and genetic integrity of human preimplantation embryos from rescue in-vitro matured MII stage oocytes as well as in vivo matured oocytes. 541 rescue in-vitro matured oocytes as case as well as 659 in-vivo matured oocytes as control were used for the developmental assay. Finally, 121 cleaved embryos with good quality were analyzed by FISH technique for the detection of chromosomes X, Y, 13, 15, 16, 18, 21 and 22. The fertilization rates were 61.62% and 61.76% in case and control groups, respectively. Also, embryo formation rates of 89.1% vs. 92.2% were recorded for case and control groups, respectively. Good quality embryos on day 3 were 62.54% in case and 68.36% in control groups. There were insignificant differences in fertilization, embryo formation and quality between the groups. Total abnormality in 35 of the 60 embryos was 58.5% in case and 62.3% in control (p < 0.05). There were significant differences between aneuploidy rates of embryos using only sex chromosome preimplantation genetic screening (PGS) and sex chromosome in combination with autosomal chromosomes PGS in case (58.5% vs 28.3%, p = 0.000) and control groups (62.3% vs 21.3%; p = 0.000). The results demonstrated that a high proportion of good quality embryos were aneuploid in both patient groups with no obvious increase in aneuploidies as a result of rescue IVM application. Furthermore, the morphological characteristics of embryos do not completely consistent with chromosomal content. Despite the Rescue IVM is currently not a routine procedure in association with IVF, our finding suggested a viable option for young infertile women facing cancellation of their IVF treatment due to ovarian over-response or resistance factors as well as patients with low functional ovarian reserve considering good quality of embryos from rescue IVM-MII oocytes.

The importance of adrenal hypoandrogenism in infertile women with low functional ovarian reserve: a case study of associated adrenal insufficiency.

BackgroundLow testosterone (T), whether due to ovarian and/or adrenal insufficiency, usually results in poor follicle maturation at small growing follicle stages. The consequence is a phenotype of low functional ovarian reserve (LFOR), characterized by poor granulosa cell mass, low anti-Müllerian hormone and estradiol but rising follicle stimulating hormone. Such hypoandrogenism can be of ovarian and/or adrenal origin. Dehydroepiandrosterone sulfate (DHEAS) is exclusively produced by adrenals and, therefore, reflects adrenal androgen production in the zona reticularis. We here determined in a case study of infertile women with LFOR the presence of adrenal hypoandrogenism, its effects on ovarian function, and the possibility of presence of concomitant adrenal insufficiency (AI), thus reflecting insufficiency of all three adrenal cortical zonae.MethodsWe searched our center’s anonymized electronic research database for women with LFOR, who were also characterized by peripheral adrenal hypoandrogenemia (total testosterone < 16.9 ng/dL) and low DHEAS (<76.0 μg/dL). Among 225 women with LFOR, we identified 29 (12.9 %). The adrenal function of so identified women were further investigated with morning cortisol and ACTH levels and/or standard ACTH stimulation tests. We also determined the prevalence of classical AI (insufficiency glucocorticoid production by zona fasciculata) in hypoandrogenic women with LFOR, and impact of adrenal hypoandrogenism on ovaries.ResultsAmong 14/28 women with adrenal hypoandrogenism due to insufficiency of the zona reticularis available for follow up, 4 (28.6 %) also demonstrated previously unrecognized classical primary, secondary or tertiary AI due to insufficiency of the zona fasciculata. An additional patient with presenting diagnosis of seemingly primary ovarian insufficiency (POI), demonstrated extremely low T and DHEAS levels, a diagnosis of Addison’s disease, and was on glucocorticoid but not androgen supplementation. As her dramatic improvement in ovarian function criteria after androgen supplementation confirmed, her correct diagnosis, therefore, was actually secondary ovarian insufficiency (SOI) due to adrenal hypoandrogenism.ConclusionsWomen with LFOR, characterized by low T and DHEAS, are also at risk for AI, while women with AI may be at risk for adrenal induced hypoandrogenism and, therefore, SOI. A currently undetermined percentage of POI patients actually are, likely, affected by SOI, a for prognostic reasons highly significant difference in diagnosis.

Associations between peripheral androgens and cortisol in infertile women

Testosterone has in recent years been proven essential for normal growth and maturation of small growing follicles. Concomitantly, low functional ovarian reserve (LFOR), characterized by a small growing follicle pool, has been associated with low testosterone levels, which can be of ovarian and/or adrenal origin. In this study we, therefore, investigated whether peripheral sex steroid precursors and testosterone levels potentially reflect on adrenal function. In a retrospective cohort study of 355 consecutive infertile women, who presented to an academically affiliated fertility center in New York City, we investigated in a series of statistical models whether low peripheral sex steroid precursors and testosterone are associated with peripheral cortisol (C) levels, reflecting adrenal function. To determine potential correlations, we investigated the dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), androstenedione (AD), total testosterone (TT), free testosterone (FT); sex hormone binding globulin (SHBG), anti-Müllerian hormone (AMH), thyroid stimulating hormone (TSH) and C in a series of multivariate and logistic regression analyses, utilizing C either as a continuous variable or with cut off <5.0μg/dL, and TT only as a continuous variable. Practically all models demonstrated significant predictability of peripheral sex hormone precursors for C levels, with DHEA demonstrating the strongest and most consistent predictability as an individual parameter and as part of the DHEAS/DHEA ratio. We conclude that in infertile women peripheral sex hormone precursors, especially DHEA, reflect C levels and, therefore, adrenal function. In infertile women, at all ages low levels of sex hormone precursors, therefore, should be considered indications for further adrenal assessments.

Rescue in vitro maturation (IVM) of immature oocytes in stimulated cycles in women with low functional ovarian reserve (LFOR).

Rescue in vitro maturation (IVM) is currently not a routine procedure in association with in vitro fertilization (IVF). We compared in a prospectively cohort study of 10 patients with normal functional ovarian reserve (NFOR) and of 25 with low functional ovarian reserve (LFOR), defined by abnormally high FSH and/or abnormally low AMH levels), IVM dynamics of immature oocytes. Following controlled ovarian hyperstimulation in IVF cycles, only immature oocytes underwent rescue IVM (for up to 48 h). Oocyte maturation dynamics, fertilization rates, embryo development, and pregnancy rates were then compared between NFOR and LFOR patients. Though proportion of MI and GV oocytes reaching MII stages within 48 h and rate of maturation of MI oocytes did not differ, in women with LFOR significantly more GV oocytes reached MII stage within 24 h (30.4 vs. 66.9 %; P = 0.013), while fertilization rates and embryo generation numbers were similar between both groups. Rescue IVM, thus, produced 1.5 additional embryos for transfer in women with LFOR and 1.6 in patients with NFOR, a highly significant difference in relative improvement in available embryo numbers for LFOR (+60.0 %) and NFOR women (+16.5 %). Rescue IVM, thus, not only demonstrates different time dynamics between women with LFOR and NFOR but also disproportionate efficacy in improving available embryo numbers for transfer in favor of LFOR patients. 1/7 patients, who reached embryo transfer with only embryos produced via rescue IVF conceived and delivered, proving that rescue IVF in women with LFOR also improves pregnancy and delivery chances. Because of the small number of embryos LFOR patients produce, every additional embryo is of considerable potential clinical significance for them, suggesting that rescue IVM in women with LFOR should become routine practice.

Live-birth rates in very poor prognosis patients, who are defined as poor responders under the Bologna criteria, with nonelective single embryo, two-embryo, and three or more embryos transferred

To determine live-birth rates (LBRs) at various ages in very poor prognosis patients, who are defined as poor responders under the Bologna criteria. Retrospective cohort study. Academically affiliated private fertility center. Among 483 patients, who under the Bologna criteria (three or fewer oocytes, >40 years of age, and/or antimüllerian hormone [AMH] <1.1 ng/mL [2/3 criteria minimum]) were poor responders, 278 (381 fresh IVF cycles) qualified for the study because they had at least one embryo on day 3 for transfer. IVF cycles in women with low functional ovarian reserve, involving androgen and CoQ10 supplementation and ovarian stimulation with daily gonadotropin dosages of 300-450 IU of FSH and 150 IU of hMG in microdose agonist cycles. Age-specific LBRs per ET. Ages did not differ between nonelective (ne) single ET (SET), ne2-ET, and ne ≥ 3-ET cycles (41.3 ± 3.9, 41.7 ± 3.1, and 42.4 ± 2.1 years, respectively). Patients with neSETs demonstrated significantly lower AMH and higher FSH levels and required higher gonadotropin dosages than ne2-ET and ne ≥ 3-ET patients. LBRs declined with age. Above age 42, three or more embryos are required to achieve reasonable LBRs and two or more to avoid futility under American Society for Reproductive Medicine (ASRM) guidelines. Very poor prognosis patients can still achieve acceptable pregnancy rates at least till their mid-40s if they reach ET. The degree to which egg donation is emphasized as the only treatment option in such patients, therefore, requires reconsideration. Above age 42, at least two, and preferably three embryos, are however required to exceed futility, as defined by ASRM.

Les acides nucléiques circulants et infertilité

Circulating nucleic acids (cell-free DNA and microRNAs) have for particularity to be easily detectable in the biological fluids of the body. Therefore, they constitute biomarkers of interest in female and male infertility care. Indeed, in female, they can be used to detect ovarian reserve disorders (polycystic ovary syndrome and low functional ovarian reserve) as well as to assess follicular microenvironment quality. Moreover, in men, their expression levels can vary in case of spermatogenesis abnormalities. Finally, circulating nucleic acids have also the ability to predict successfully the quality of in vitro embryo development. Their multiple contributions during assisted reproductive technology (ART) make of them biomarkers of interest, for the development of new diagnostic and/or prognostic tests, applied to our specialty. Circulating nucleic acids would so offer the possibility of personalized medical care for infertile couples in ART.

Cell-free DNA in Human Follicular Microenvironment: New Prognostic Biomarker to Predict in vitro Fertilization Outcomes.

Cell-free DNA (cfDNA) fragments, detected in blood and in other biological fluids, are released from apoptotic and/or necrotic cells. CfDNA is currently used as biomarker for the detection of many diseases such as some cancers and gynecological and obstetrics disorders. In this study, we investigated if cfDNA levels in follicular fluid (FF) samples from in vitro fertilization (IVF) patients, could be related to their ovarian reserve status, controlled ovarian stimulation (COS) protocols and IVF outcomes. Therefore, 117 FF samples were collected from women (n = 117) undergoing IVF/Intra-cytoplasmic sperm injection (ICSI) procedure and cfDNA concentration was quantified by ALU-quantitative PCR. We found that cfDNA level was significantly higher in FF samples from patients with ovarian reserve disorders (low functional ovarian reserve or polycystic ovary syndrome) than from patients with normal ovarian reserve (2.7 ± 2.7 ng/μl versus 1.7 ± 2.3 ng/μl, respectively, p = 0.03). Likewise, FF cfDNA levels were significant more elevated in women who received long ovarian stimulation (> 10 days) or high total dose of gonadotropins (≥ 3000 IU/l) than in women who received short stimulation duration (7–10 days) or total dose of gonadotropins < 3000 IU/l (2.4 ± 2.8 ng/μl versus 1.5 ± 1.9 ng/μl, p = 0.008; 2.2 ± 2.3 ng/μl versus 1.5 ± 2.1 ng/μl, p = 0.01, respectively). Finally, FF cfDNA level was an independent and significant predictive factor for pregnancy outcome (adjusted odds ratio = 0.69 [0.5; 0.96], p = 0.03). In multivariate analysis, the Receiving Operator Curve (ROC) analysis showed that the performance of FF cfDNA in predicting clinical pregnancy reached 0.73 [0.66–0.87] with 88% specificity and 60% sensitivity. CfDNA might constitute a promising biomarker of follicular micro-environment quality which could be used to predict IVF prognosis and to enhance female infertility management.

Relative importance of AMH and androgens changes with aging among non-obese women with polycystic ovary syndrome.

BackgroundTo assess the changes in phenotypes and endocrine profiles of women with polycystic ovary syndrome (PCOS) with advancing age.MethodsIn a cross-sectional study conducted at a private tertiary fertility clinical and research center we identified anonymized electronic records of 37 women who had presented with a prior diagnosis of PCOS. They were stratified as younger (<35 years) and older (≥40 years). As controls, we identified 43 women with age-specific low functional ovarian reserve and 14 young women with normal functional ovarian reserve. Endocrine profiles for each group were evaluated based on total (TT) and free testosterone (FT), anti-Müllerian hormone (AMH) and sex hormone binding globulin (SHBG).ResultsPatients including those with PCOS were mostly non-obese, evidenced by normal BMIs (21.6 ± 6.0) with no differences between study groups. Young PCOS patients presented with a typical pattern of significant hyperandrogenemia and elevated AMH in comparison to young women with normal functional ovarian reserve [TT 44.0 (32.9–58.7) vs. 23.9 (20.3–28.1) ng/dL, (P<0.05); and AMH 7.7 (6.2–9.1) vs. 2.5 (2.0–3.0) ng/mL, (P<0.05)]. With advancing age, hyperandrogenemia in PCOS diminished in comparison to young women with normal functional ovarian reserve, resulting in similar TT levels [28.6 (19.7–37.5) vs. 23.9 (20.3–28.1) ng/dL]. Though also declining, AMH remained significantly elevated in older PCOS women in comparison to young women with normal functional ovarian reserve [4.0 (2.7–5.2) vs. 2.5 (2.0–3.0) ng/mL, (P<0.05)]. Patients with low functional ovarian reserve demonstrated significantly lower AMH at both young and older ages compared to women with normal functional ovarian reserve (P<0.05 for both). However, among patients with low functional ovarian reserve no differences were observed at young compared to older ages in TT [17.6 (12.9–24.1) vs. 18.1 (13.6–24.1) ng/dL)] and AMH [0.4 (0.3–0.6) vs. 0.3 (0.2–0.5) ng/mL]. SHBG did not differ significantly between groups but trended opposite to testosterone.ConclusionsThe PCOS population predominantly consisted of non-obese phenotype at both young and advanced ages. This suggests that patients with “classical” obese PCOS phenotype rarely reach tertiary infertility care, while non-obese PCOS patients may be more resistant to lower levels of infertility treatments. PCOS patients also demonstrate more precipitous declines in testosterone then AMH with advancing age. These data support incorporation of AMH as diagnostic criterion for PCOS regardless of age, and imply that testosterone should not be relied upon in the diagnosis of PCOS in older women.

The impact of thyroid function and thyroid autoimmunity on embryo quality in women with low functional ovarian reserve: a case-control study.

BackgroundWomen with hyper-and hypothyroidism are at increased risk for infertility and adverse pregnancy outcomes. Whether in women considered euthyroid thyroid function (TSH values) and thyroid autoimmunity (thyroid antibodies) influence in vitro fertilization (IVF) cycle outcome has, however, remained controversial. Any such effect should be easily visible in women with low functional ovarian reserve (LFOR) and thus small oocyte and embryo numbers.MethodsWe evaluated the relationship between TSH levels and embryo quality in euthyroid women with LFOR undergoing IVF. Mean age for the study population was 39.9 ± 4.6 years. Embryo quality was assessed in 431 embryos from 98 first IVF cycles according to TSH levels (with cut-off 2.5μIU/mL), and to presence versus absence of thyroid autoantibodies.ResultsMean Anti Mullerian hormone (AMH) was 0.8 ± 0.8 ng/mL and mean TSH was 1.8 ± 0.9 μIU/mL. Comparable embryo quality was observed in women with TSH ≤ and >2.5μIU/mL. TPO antibodies significantly affected embryo quality in women with low-normal TSH levels (P = 0.045). In women with high-normal TSH levels, increasing TSH had a negative impact on embryo quality (P = 0.027). A trend towards impaired embryo quality with TPO antibodies was also observed in these patients (p = 0.057).ConclusionsTPO antibodies affect embryo quality in euthyroid women with low-normal TSH ≤2.5 μIU/mL. In women with high-normal TSH levels, increasing TSH levels, and possibly TPO antibodies, appear to impair embryo quality. These results suggest that the negative impact of thyroid autoimmunity becomes apparent, once thyroid hormone function is optimized.

Is there an androgen level threshold for aneuploidy risk in infertile women?

BackgroundLow functional ovarian reserve (LFOR) has been associated with hypoandrogenemia and increased embryo aneuploidy, while androgen supplementation has been reported to improve aneuploidy rates. We, therefore, assessed whether in infertile women undergoing in vitro fertilization (IVF) androgen concentrations are associated with aneuploidy rates.MethodsThis study was performed in 2 academically affiliated fertility centers in New York City and an academically affiliated steroid chemistry laboratory in Utah. Androgen concentrations were measured in blinded fashion from 84 infertile women (age 40.3 +/− 2.4 years) at New York University (NYU), using a validated LC-MS/MS method, in cryopreserved serum samples of patients who had undergone IVF with concomitant preimplantation genetic screening (PGS), utilizing a 24-chromosome platform. The Center for Human Reproduction (CHR) provided plasma samples of 100 historical controls (ages 38.6+/−5.0 years) undergoing IVF without PGS. Statistical comparisons were made of androgen concentrations, and of associations between androgen concentrations and embryo aneuploidy.ResultsWomen undergoing IVF + PGS at NYU revealed no association between embryo aneuploidy and androgen concentrations but demonstrated significantly lower androgen concentrations than the 100 control patients from CHR,ConclusionsThough this study revealed no association between androgen levels and embryo ploidy, the extremely low androgen levels in the NYU study group raise the possibility of a threshold effect below which testosterone no longer affects aneuploidy. Before an androgen effect on embryo ploidy can be completely ruled out, a patient population with more normal androgen levels has to be investigated.

Genetics of androgen metabolism in women with infertility and hypoandrogenism.

Hypoandrogenism in women with low functional ovarian reserve (LFOR, defined as an abnormally low number of small growing follicles) adversely affects fertility. The androgen precursor dehydroepiandrosterone (DHEA) is increasingly used to supplement treatment protocols in women with LFOR undergoing in vitro fertilization. Due to differences in androgen metabolism, however, responses to DHEA supplementation vary between patients. In addition to overall declines in steroidogenic capacity with advancing age, genetic factors, which result in altered expression or enzymatic function of key steroidogenic proteins or their upstream regulators, might further exacerbate variations in the conversion of DHEA to testosterone. In this Review, we discuss in vitro studies and animal models of polymorphisms and gene mutations that affect the conversion of DHEA to testosterone and attempt to elucidate how these variations affect female hormone profiles. We also discuss treatment options that modulate levels of testosterone by targeting the expression of steroidogenic genes. Common variants in genes encoding DHEA sulphotransferase, aromatase, steroid 5α-reductase, androgen receptor, sex-hormone binding globulin, fragile X mental retardation protein and breast cancer type 1 susceptibility protein have been implicated in androgen metabolism and, therefore, can affect levels of androgens in women. Short of screening for all potential genetic variants, hormonal assessments of patients with low testosterone levels after DHEA supplementation facilitate identification of underlying genetic defects. The genetic predisposition of patients can then be used to design individualized fertility treatments.

Differences in AMH assessments between standard and new high sensitivity AMH assays and impact on prediction of IVF outcomes

Since women with AMH levels below current limits of detection often still respond to ovulation induction, low limits of currently widely utilized anti-Müllerian hormone (AMH) assays (∼0.16ng/mL) are clinically limiting in women with low functional ovarian reserve (LFOR). We, therefore, assessed whether a mores sensitive AMH assay may improve prediction of treatment outcomes in LFOR.