Fred Moy

Impairment of BRCA1-Related DNA Double-Strand Break Repair Leads to Ovarian Aging in Mice and Humans

DNA double-strand break repair has a central role in oocyte aging. Preserving Fertility Breeds Flexibility Last month, the U.K. Office for National Statistics reported that, in 2010, ~48% of infants were born to mothers 30 years and older, a level not seen since 1946—just after the end of World War II. Delaying childbearing can allow women flexibility with respect to career options. But unlike many somatic tissues, the female germline ages early, with reproductive capacity beginning to diminish after young adulthood. Attempts to stem oocyte aging and preserve fertility will depend on finely characterizing the molecular mechanisms behind the aging process in the female reproductive system. Now, Titus et al. provide evidence for a new mechanism to explain age-related oocyte dysfunction. The authors showed that double-stranded breaks (DSBs) in DNA—which are essential for normal development—accumulate with age and contribute to reproductive aging in mice and women. In single mouse and human oocytes, the expression of DSB repair genes BRCA1, MRE11, RAD51, and ATM declined with age. Thus DSBs likely collect in the oocyte genome because of age-related missteps in DSB repair, which stimulate apoptosis and diminishes ovarian reserve. Indeed, in Brca1-deficient mice, numbers of primordial follicles—immature primary oocytes—were decreased, DSBs were increased, and reproductive capacity was impaired relative to wild-type mice. Using RNA interference in mouse oocytes, the authors showed that inhibition of Brca1, MRE11, RAD51, and, in turn, ATM expression increased DSBs and reduced oocyte survival. The authors then measured serum concentrations of anti-Müllerian hormone—a measure of fertility—in young women with germline BRCA1 mutations versus controls and showed that ovarian reserve was compromised in the latter group. Together, these findings show that the efficiency of DNA DSB repair is a crucial determinant of oocyte loss. The discovery of therapies that target this pathway might help to enhance the duration of ovarian function. The underlying mechanism behind age-induced wastage of the human ovarian follicle reserve is unknown. We identify impaired ATM (ataxia-telangiectasia mutated)–mediated DNA double-strand break (DSB) repair as a cause of aging in mouse and human oocytes. We show that DSBs accumulate in primordial follicles with age. In parallel, expression of key DNA DSB repair genes BRCA1, MRE11, Rad51, and ATM, but not BRCA2, declines in single mouse and human oocytes. In Brca1-deficient mice, reproductive capacity was impaired, primordial follicle counts were lower, and DSBs were increased in remaining follicles with age relative to wild-type mice. Furthermore, oocyte-specific knockdown of Brca1, MRE11, Rad51, and ATM expression increased DSBs and reduced survival, whereas Brca1 overexpression enhanced both parameters. Likewise, ovarian reserve was impaired in young women with germline BRCA1 mutations compared to controls as determined by serum concentrations of anti-Müllerian hormone. These data implicate DNA DSB repair efficiency as an important determinant of oocyte aging in women.

Endometriosis: a stage by stage analysis--the role of in vitro fertilization.

OBJECTIVE To investigate the impact of endometriosis stage on IVF. PATIENTS A retrospective study of 214 patients diagnosed with endometriosis who underwent 360 cycles of IVF at The New York Hospital-Cornell Medical Center. Meanwhile, 111 pure mechanical (tubal) infertility patients treated in 160 cycles at the same time were designated as the control group for comparison. MAIN OUTCOME MEASURES Patients hormone and semen profiles, hormonal response and outcome to stimulation, as well as the outcome of pregnancy, abortion, and delivery rate were analyzed. RESULTS No differences in the pregnancy outcome between the endometriosis and control groups were noted when compared among the subgroups of pure endometriosis, endometriosis plus tubal factor, endometriosis plus others (primarily endometriosis plus male factor), and control. Comparing the outcomes in pure endometriosis cases by staging, we could not find any discrepancies in terms of pregnancy rates (PRs) according to the severity of the disease. The addition of GnRH analogur down-regulation to gonadotropin stimulation resulted in an increase in PR. A relatively high delivery rate (38.9% per cycle, 41.9% per retrieval, and 43.2% per transfer) was achieved when the pure endometriosis patients were treated with concomitant leuprolide acetate down-regulation and gonadotropin. CONCLUSION We have observed that pregnancy outcome in patients with endometriosis was not different than the outcome for patients with mechanical (tubal) infertility. There were no differences in PRs by stage of endometriosis.

Current Progress on Understanding MicroRNAs in Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is an aggressive grade IV astrocytoma with a 1-year median survival rate despite current treatment modalities. A thorough understanding of the vast genetic aberrations and signaling pathways involved in gliomagenesis as well as heterogeneous clinicopathological presentation remains elusive. The recent discovery of microRNAs (miRs) and their capability of simultaneously regulating multiple downstream genes may play a key role in explaining the complex mechanisms underlying GBM formation. miRs are 19 to 25 nucleotide non-protein-coding small RNA molecules involved in the suppression of mRNA translation. This review will summarize and discuss the most recent findings regarding miRs in GBM including downstream targets, functional effects, and therapeutic potentials. Specifically discussed miRs include miR-7, miR-9/miR-9*, miR-10a/miR-10a*/miR-10b, miR-15b, miR-17-92, miR-21, miR-26a, miR-34a, miR-93, miR-101, miR-124, miR-125a, miR-125b, miR-128, miR-137, miR-146b-5p, miR-153, miR-181a/miR-181b, miR-196a/miR-196b, miR-218, miR-221/miR-222, miR-296, miR-302-367, miR-326, miR-381, miR-451, and let-7a. In addition to gene regulatory roles, miRs have demonstrated significant diagnostic, prognostic, and therapeutic potential. These small molecules may both help in the understanding of GBM and in developing new therapeutic options.

Value of Early Referral to Fertility Preservation in Young Women With Breast Cancer

PURPOSE To determine whether early referral to reproductive specialists improves fertility preservation (FP) outcomes and reduces delay in adjuvant treatment in young women with breast cancer. PATIENTS AND METHODS A secondary analysis of a prospective database of patients with breast cancer undergoing ovarian stimulation (OS) for FP by oocyte or embryo cryopreservation was performed. RESULTS Of the 154 patients, 93 met the inclusion criteria (mean age, 35.2 ± 4.4 years). Thirty-five of the 93 patients were referred before breast surgery (PreS), and 58 patients were referred after surgery (PostS). The time periods from initial diagnosis (ID) to initiation of OS (42.6 ± 27.7 days for PreS v 71.9 ± 30.7 days for PostS; P < .001) and from ID to initiation of chemotherapy (83.9 ± 24.3 days for PreS v 107.8 ± 42.9 days for PostS; P = .045) were significantly shorter for the PreS group versus the PostS group. Nine (25.7%) of 35 patients in the PreS group versus one (1.7%) of 58 patients in the PostS group were able to undergo two FP cycles (P < .001), resulting in an increased yield of oocytes in the PreS group (18.2% [93 of 511 oocytes] v 0.6% [five of 800 oocytes], respectively; P < .001) and embryos (17.2% [40 of 233 embryos] v 0.6% [two of 357 embryos], respectively; P < .001). Patients who had an oocyte retrieval within 5 weeks of the surgery were able to complete a second cycle within 9 weeks of the surgery. CONCLUSION FP referral before breast surgery enables earlier initiation of cryopreservation cycles and chemotherapy and, when appropriate, multiple FP cycles. Women who can undergo multiple cycles may be at advantage for FP because of a larger number of oocytes or embryos cryopreserved. This is the first study demonstrating the benefit of early FP referral in patients with cancer.

Salmon Aquaculture and Antimicrobial Resistance in the Marine Environment

Antimicrobials used in salmon aquaculture pass into the marine environment. This could have negative impacts on marine environmental biodiversity, and on terrestrial animal and human health as a result of selection for bacteria containing antimicrobial resistance genes. We therefore measured the numbers of culturable bacteria and antimicrobial-resistant bacteria in marine sediments in the Calbuco Archipelago, Chile, over 12-month period at a salmon aquaculture site approximately 20 m from a salmon farm and at a control site 8 km distant without observable aquaculture activities. Three antimicrobials extensively used in Chilean salmon aquaculture (oxytetracycline, oxolinic acid, and florfenicol) were studied. Although none of these antimicrobials was detected in sediments from either site, traces of flumequine, a fluoroquinolone antimicrobial also widely used in Chile, were present in sediments from both sites during this period. There were significant increases in bacterial numbers and antimicrobial-resistant fractions to oxytetracycline, oxolinic acid, and florfenicol in sediments from the aquaculture site compared to those from the control site. Interestingly, there were similar numbers of presumably plasmid-mediated resistance genes for oxytetracycline, oxolinic acid and florfenicol in unselected marine bacteria isolated from both aquaculture and control sites. These preliminary findings in one location may suggest that the current use of large amounts of antimicrobials in Chilean aquaculture has the potential to select for antimicrobial-resistant bacteria in marine sediments.

Fertility Preservation Success Subsequent to Concurrent Aromatase Inhibitor Treatment and Ovarian Stimulation in Women With Breast Cancer

PURPOSE We have previously reported an approach to ovarian stimulation for the purpose of fertility preservation (FP) in women with breast cancer via embryo freezing with the concurrent use of letrozole. The aim of this study was to provide the pregnancy and FP outcomes when embryos generated with the same protocol are used. PATIENTS AND METHODS In all, 131 women with stage ≤ 3 breast cancer underwent ovarian stimulation and received concurrent letrozole 5 mg per day before receiving adjuvant chemotherapy and cryopreserving embryos. RESULTS Thirty-three of the 131 women underwent 40 attempts to transfer embryos to their own uterus (n = 18) or via the use of a gestational carrier (n = 22) at a mean age of 41.5 ± 4.3 years with a median 5.25 years after embryo cryopreservation. The overall live birth rate per embryo transfer was similar to the US national mean among infertile women of a similar age undergoing in vitro fertilization-embryo transfer (45.0 v 38.2; P = .2). Seven (38.8%) of the 18 pregnancies were twins with no higher-order pregnancies being encountered. No fetal anomalies or malformations were reported in 25 children after a mean follow-up of 40.4 ± 26.4 months. Seventeen of the 33 women attempting pregnancy had at least one child, translating into an FP rate of 51.5% per attempting woman. CONCLUSION Embryo cryopreservation after ovarian stimulation with the letrozole and follicle-stimulating hormone protocol preserves fertility in women with breast cancer and results in pregnancy rates comparable to those expected in a noncancer population undergoing in vitro fertilization.

Landscape features associated with lyme disease risk in a suburban residential environment

The landscape features of residential properties within two communities were studied in relation to the abundance of the tick vector Ixodes scapularis. Habitat types of 400 properties, located in a Lyme disease endemic area of Westchester Co., New York, USA, were categorized into lawn, ornamental, ecotone, woods, and stone wall as measured from aerial photographs and sampled for nymphal-stage ticks. Logistic regression results indicate that presence or absence of ticks is influenced by the proportion of either lawn or woodland, and total woodland area. Poisson regression results indicate the abundance of nymphs is negatively associated with proportion, area, and patch frequency of lawn, and positively associated with proportion, area, and patch frequency of woodland. Predictions of tick presence and abundance from landscape features at the scale of individual property is useful for implementing disease prevention measures.

Genetic and epigenetic characteristics of ICSI children

The outcome of pregnancy and the developmental wellbeing of children conceived from 12,866 consecutive intracytoplasmic sperm injection (ICSI) cycles was assessed. A total of 3277 couples delivered 5891 neonates. There was a higher than normal incidence of de-novo chromosomal abnormalities in a small sample of ICSI offspring. Controlling for maternal age showed that the incidence of low birth weight and gestational length were comparable with the naturally conceived counterpart. Rates of malformation in ICSI offspring ranged from 3.5 to 6.2%. At 3 years of age (n = 811), the proportion of children at risk for developmental delays was 10.4% in ICSI and 10.7% in IVF singletons. However, high order gestations were characterized by 19.4% of the children having compromised development. Epigenetic analysis of assisted reproductive technique conceptuses found minor imprinted gene expression imbalances. ICSI offspring presented with genetic defects that were inherited or arose de novo. Obstetric and neonatal outcomes of singleton pregnancies appeared to be dependent upon maternal age. ICSI and IVF appeared to exert a negative effect on the wellbeing of offspring mainly because of the association with multiple gestations. All assisted reproduction procedures should be monitored for the eventual effect of environmental aggressors on offspring epigenesis.

Safety and feasibility of performing two consecutive ovarian stimulation cycles with the use of letrozole-gonadotropin protocol for fertility preservation in breast cancer patients

OBJECTIVE To investigate the safety and feasibility of performing two consecutive ovarian stimulation cycles with the use of letrozole protocol for fertility preservation in breast cancer patients. DESIGN Retrospective cohort study. SETTING Academic fertility preservation center. PATIENT(S) Seventy-eight women ≤ 45 years, diagnosed with stage ≤ 3 breast cancer, who desired fertility preservation. INTERVENTION(S) Two consecutive cycles versus a single ovarian stimulation cycle with a letrozole-follicle-stimulating hormone (FSH) protocol. MAIN OUTCOME MEASURE(S) Embryo or oocyte cryopreservation outcomes, time interval from surgery to chemotherapy, and breast cancer recurrence rates. RESULT(S) Sixty-one patients underwent single-cycle stimulation and 17 received two stimulation cycles. The mean total number of oocytes harvested (16.1 ± 13.2 vs. 9.1 ± 5.2) and embryos generated (6.4 ± 2.9 vs. 3.7 ± 3.1) were statistically significantly higher in patients who underwent two cycles versus one cycle. The time interval from surgery to chemotherapy was similar between the two-cycle and single-cycle groups (63.7 ± 7.7 vs. 58.0 ± 12.1 days). After a mean follow-up interval of 58.5 ± 13.6 months, the recurrence rates were similar between the two-cycle (0 of 17) and single-cycle (2 of 49) patients. CONCLUSION(S) It appears to be safe and feasible to perform two consecutive ovarian stimulation cycles to increase the oocyte/embryo yield for fertility preservation.

Determinants of access to fertility preservation in women with breast cancer

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 Secondary analysis. SETTING Academic medical centers. PATIENT(S) 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) None. MAIN OUTCOME MEASURE(S) Factors favoring early referrals. RESULT(S) 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) 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.