Faculty Bibliography

PURPOSE/OBJECTIVE:To predict embryo/oocyte cryopreservation cycle (ECC) outcomes in breast cancer patients stimulated with letrozole and follicle stimulating hormone for fertility preservation based on observed anti-mullerian hormone (AMH) levels and antral follicle counts (AFC). METHODS:The correlation between AMH and AFC and ECC outcomes were analyzed retrospectively on forty one women with breast cancer before adjuvant treatment. RESULTS:AMH and AFC had a stronger correlation with the total number of oocytes and the number of mature oocytes than age, FSH, and inhibin B. Subjects were evaluated by the number of mature oocytes retrieved to create cutoff points of AMH level, which identified 1.2 ng/mL as a potential value. Seven of 18 patients with AMH levels ≤1.2 ng/mL had low response versus none of 23 with >1.2 ng/mL, (p = 0.001). CONCLUSIONS:AMH is the most reliable serum marker of ECC outcomes, together with AFC as a biophysical marker, in breast cancer patients. Low response is highly likely when the AMH level is ≤1.2 ng/mL.

OBJECTIVE:To report an emergency approach of random-start controlled ovarian hyperstimulation (COH) in the late follicular or luteal phase of the menstrual cycle for embryo cryopreservation in patients with cancer. DESIGN/METHODS:Case series. SETTING/METHODS:Academic tertiary referral centers. PATIENT(S)/METHODS:Three patients with a diagnosis of breast cancer requiring emergency fertility preservation in the late follicular or luteal phase of the menstrual cycle. INTERVENTION(S)/METHODS:After baseline pelvic ultrasound and hormonal evaluation, random-start COH was commenced immediately on menstrual cycle days 11, 14, or 17 with use of letrozole 2.5 mg/d and recombinant FSH 150 to 300 IU/d. Gonadotropin-releasing hormone antagonist was administered to prevent ovulation in all cases. Ovulation was triggered with either 250 μg of recombinant hCG or 10,000 IU of urinary hCG. MAIN OUTCOME MEASURE(S)/METHODS:Number of oocytes harvested, maturity and fertilization rates, number of embryos frozen. RESULT(S)/RESULTS:Nine to 17 oocytes were harvested, resulting in the freezing of seven to 10 embryos with the mean maturity and fertilization rates of 58.8% to 77.7% and 69.2% to 87.5%, respectively. CONCLUSION(S)/CONCLUSIONS:In an emergent setting, ovarian stimulation can be started at a random cycle date for the purpose of fertility preservation without compromising fertilization rates in letrozole cycles.

Ovarian transplantation is one of the key approaches to restoring fertility in women who became menopausal as a result of cancer treatments. A major limitation of human ovarian transplants is massive follicular loss during revascularization. Here we investigated whether sphingosine-1-phosphate or its receptor agonists could enhance neoangiogenesis and follicle survival in ovarian transplants in a xenograft model. Human ovarian tissue xenografts in severe-combined-immunodeficient mice were treated with sphingosine-1-phosphate, its analogs, or vehicle for 1-10 days. We found that sphingosine-1-phosphate treatment increased vascular density in ovarian transplants significantly whereas FTY720 and SEW2871 had the opposite effect. In addition, sphingosine-1-phosphate accelerated the angiogenic process compared to vehicle-treated controls. Furthermore, sphingosine-1-phosphate treatment was associated with a significant proliferation of ovarian stromal cell as well as reduced necrosis and tissue hypoxia compared to the vehicle-treated controls. This resulted in a significantly lower percentage of apoptotic follicles in sphingosine-1-phosphate-treated transplants. We conclude that while sphingosine-1-phosphate promotes neoangiogenesis in ovarian transplants and reduces ischemic reperfusion injury, sphingosine-1-phosphate receptor agonists appear to functionally antagonize this process. Sphingosine-1-phosphate holds great promise to clinically enhance the survival and longevity of human autologous ovarian transplants.

OBJECTIVE:To evaluate socioeconomic, demographic, and medical factors that influence the referral pattern-either before cancer treatment for fertility preservation (FP, early referral) or post-chemotherapy for assisted reproductive technology (PCART, delayed referral)-in women with breast cancer. DESIGN/METHODS:Secondary analysis. SETTING/METHODS:Academic medical centers. PATIENT(S)/METHODS:Three hundred fourteen patients with breast cancer who were counseled for FP (n=218) or PCART (n=96) from June 1999 to July 2009. INTERVENTION(S)/METHODS:None. MAIN OUTCOME MEASURE(S)/METHODS:Factors favoring early referrals. RESULT(S)/RESULTS:Mean age at diagnosis was higher in FP vs. PCART (35.3±4.5 years vs. 33.9±4.7 years). Ninety percent presented with cancer stage 1 or 2. From 2000 to 2009 the proportion of referrals for FP increased continually. In 2009, nearly all (95.5%) were for FP. The majority (63.8%) was referred from an academic center. Patients with a family history of breast cancer were more likely to consult for FP (75.2% vs. 64.3% without). There was no association with occupation, income, race, ethnicity, obstetric history, and prior infertility treatment. Only 22.9% of those counseled in PCART, compared with 45.0% in the FP group, proceeded with a procedure. CONCLUSION(S)/CONCLUSIONS:There has been an increasing trend within the last 10 years for early referral of breast cancer patients to FP. Factors favoring early referrals are older age, early-stage cancer, family history of breast cancer, and academic center involvement. Those seen before cancer treatment are more likely to receive an intervention.

c-Jun N-terminal kinase (JNK) pathway has been shown to be essential for cell cycle progression and mitosis. We previously showed that this pathway is activated in mitotic granulosa cells of follicles from transitional to antral stages. In this study, we, therefore, aimed to investigate whether this signaling pathway has any effect on in-vitro growth of murine preantral follicles and granulosa cell cycle control. Two structurally different pharmacologic JNK inhibitors, SP600125 and AS601245, were used in the experiments. First their inhibitory concentrations were determined in granulosa cells by Western blot analysis. Then preantral follicles isolated from immature and adult C57BL/6 mice were cultured in matrigel and standard culture plates for 6 days with these inhibitors. Spontaneously immortalized rat granulosa cells (SIGCs) were first synchronized at G1/S and G2/M stages of cell cycle and then treated with JNK inhibitors. Cell cycle progression was analyzed with Bromodeoxyuridine (BrdU) assay and flow cytometry analysis. Both inhibitors significantly inhibited phosphorylation of c-Jun in granulosa cells at 25, 50, and 100 μmol/L concentrations. Isolated preantral follicles cultured with these inhibitors exhibited arrested growth in culture in a dose-dependent manner. Cell cycle analyses showed that both inhibitors impair the progression of cell cycle at S phase and G2/M transition of granulosa cells. These results suggest that JNK pathway is essential for in vitro growth of preantral follicle growth and regulates both S phase and G2/M stages of cell cycle in granulosa cells.

Aromatase inhibitors can be utilized to minimize oestrogen exposure in breast cancer patients undergoing gonadotrophin stimulation. This retrospective-prospective study determined whether using a gonadotrophin-releasing hormone agonist (GnRHa) trigger instead of human chorionic gonadotrophin (HCG) would reduce oestrogen exposure and improve cycle outcomes in aromatase inhibitor cycles. Seventy-four breast cancer patients who desired fertility preservation, with normal ovarian reserve and < 45 years of age received letrozole 5mg/day plus recombinant FSH 150-300 IU/day for ovarian stimulation. Subjects either received HCG 5000-10,000 IU (n=47) or leuprolide acetate 1mg (GnRHa, n=27) as trigger. Oestradiol measurements were repeated 4 days after the trigger and subjects were evaluated for ovarian hyperstimulation syndrome (OHSS). In the GnRHa group, oestradiol concentrations dropped significantly after the trigger than the HCG group (P=0.013) and there was a lower incidence of OHSS. GnRHa trigger resulted in a higher number and percentage of mature oocytes and a higher number of cryopreserved embryos or oocytes compared with HCG. GnRHa trigger improves outcomes by increasing the yield of mature oocytes and embryos in aromatase inhibitor cycles and also decreases the post-trigger oestradiol exposure as well as OHSS risks in women with breast cancer.

Clinical studies of oocyte cryopreservation have gained momentum within the recent years; however, no guidelines have yet been established for patient selection. This article discusses the controversial aspects of selecting candidates for oocyte cryopreservation research.

Demographic data and tumour characteristics of 18 patients (study group) diagnosed with breast cancer within 24 months of undergoing ovarian stimulation with either gonadotrophins or clomiphene citrate were evaluated and compared with similar 102 age-matched women diagnosed with breast cancer without prior infertility treatment (control group). Eight out of 17 (47.1%) patients in the study group and 35/95 (36.8%) patients in the control group had positive family history for breast cancer. Median tumour size was similar in the study and control groups (both 1.3 cm). Both groups were comparable regarding tumour histological types and oestrogen receptor, progesterone receptor and Her2/Neu expression status. Albeit not significant, stage 0 tumours were more prevalent in the study group compared with the control group (22.2% versus 10.5%), and there were no stage III tumours in the study group as opposed to 7/95 in the control group. In conclusion, breast cancer diagnosed within the first 2 years following infertility treatment is similar in tumour characteristics compared with those occurring in patients without prior infertility treatment.

Although still experimental, cryopreservation and transplantation techniques for ovarian tissue have been well described, and a number of successful human pregnancies have occurred. Ovarian cryopreservation is the only fertility preservation procedure that can be offered to prepubertal children, and when cytotoxic treatment is urgent. There are two main approaches for autotransplantation of human ovarian tissue. In the heterotopic transplantation, cortical fragments can be grafted subcutaneously at various sites whereas in orthotopic transplantation cortical pieces are transplanted into its original location. Both approaches have their own advantages and disadvantages. While natural pregnancy can occur in orthotopic transplantation, heterotopic transplantation may be indicated if the pelvis is not suitable for transplantation due to previous radiation or severe scar formation. Furthermore, tissue monitoring may be easier in the heterotopic site. In this article, we reviewed the indications, limitations, risks and transplantation techniques for ovarian tissue.