Low Estrogen States Research Articles

Kisspeptin in functional hypothalamic amenorrhea: Pathophysiology and therapeutic potential.

Functional hypothalamic amenorrhea (FHA) is one of the most common causes of secondary amenorrhea, resulting in anovulation and infertility, and is a low estrogen state that increases the risk of cardiovascular disease and impairs bone health. FHA is characterized by acquired suppression of physiological pulsatile gonadotropin-releasing hormone (GnRH) release by the hypothalamus in the absence of an identifiable structural cause, resulting in a functional hypogonadotropic hypogonadism. FHA results from either decreased energy intake and/or excessive exercise, leading to low energy availability and weight loss-often in combination with psychological stress on top of a background of genetic susceptibility. The hypothalamic neuropeptide kisspeptin is a key component of the GnRH pulse generator, tightly regulating pulsatile GnRH secretion and the downstream reproductive axis. Here, we review the physiological regulation of pulsatile GnRH secretion by hypothalamic kisspeptin neurons and how their activity is modulated by signals of energy status to affect reproductive function. We explore endocrine factors contributing to the suppression of GnRH pulsatility in the pathophysiology of FHA and how hypothalamic kisspeptin neurons likely represent a final common pathway through which these factors affect GnRH pulse generation. Finally, we discuss the therapeutic potential of kisspeptin as a novel treatment for women with FHA.

Oxidative stress plays an important role in the central regulatory mechanism of orofacial hyperalgesia under low estrogen conditions

Hyperalgesia occurs in the orofacial region of rats when estrogen levels are low, although the specific mechanism needs to be investigated further. Furthermore, oxidative stress plays an important role in the transmission of pain signals. This study aimed to explore the role of oxidative stress in orofacial hyperalgesia under low estrogen conditions. We firstly found an imbalance between oxidative and antioxidant capacity within the spinal trigeminal subnucleus caudalis (SP5C) of rats after ovariectomy (OVX), resulting in oxidative stress and then a decrease in the orofacial pain threshold. To investigate the mechanism by which oxidative stress occurs, we used virus as a tool to silence or overexpress the excitatory amino acid transporter 3 (EAAT3) gene. Further investigation revealed that the regulation of glutathione (GSH) and reactive oxygen species (ROS) can be achieved by regulating EAAT3, which in turn impacts the occurrence of oxidative stress. In summary, our findings suggest that reduced expression of EAAT3 within the SP5C of rats in the low estrogen state may decrease GSH content and increase ROS levels, resulting in oxidative stress and ultimately lead to orofacial hyperalgesia. This suggests that antioxidants could be a potential therapeutic direction for orofacial hyperalgesia under low estrogen conditions, though more research is needed to understand its mechanism.

Vaginal microbiomes of breast cancer survivors treated with aromatase inhibitors with and without vulvovaginal symptoms

Genitourinary syndrome of menopause (GSM) is the leading cause of vaginal symptoms in breast cancer survivors treated with aromatase inhibitors. However, there are currently no effective treatment options available for women with a history of breast cancer. Recent research has established that changes in the vaginal microbiome may be linked to GSM. Most studies have assessed the microbiome without accounting for the estrogen status. It remains unknown whether the vaginal microbiome differ among patients with a low estrogenic state with and without vulvovaginal symptoms. To address such research questions, our study compares the vaginal microbiomes among breast cancer survivors treated with aromatase inhibitors with and without vulvovaginal symptoms. A total of 50 breast cancer survivors treated with aromatase inhibitors were recruited, among whom 25 had vulvovaginal symptoms and 25 had no vulvovaginal symptoms. Vaginal swabs were collected. DNA extraction, followed by sequencing of the V3–V4 regions of the 16S ribosomal RNA gene, were performed. Differential abundance analysis was conducted by linear discriminant analysis effect size. Taxonomy assignment, alpha diversity and beta diversity were examined. The relative abundance of genus Sneathia and genus Gardnerella was significantly increased in vulvovaginal symptoms group with no differences in bacterial diversity and richness.

Early-life stress and ovarian hormones alter transcriptional regulation in the nucleus accumbens resulting in sex-specific responses to cocaine

Early-life stress and ovarian hormones contribute to increased female vulnerability to cocaine addiction. Here, we reveal molecular substrates in the reward area, the nucleus accumbens, through which these female-specific factors affect immediate and conditioning responses to cocaine. We find shared involvement of X chromosome inactivation-related and estrogen signaling-related gene regulation in enhanced conditioning responses following early-life stress and during the low-estrogenic state in females. Low-estrogenic females respond to acute cocaine by opening neuronal chromatin enriched for the sites of ΔFosB, a transcription factor implicated in chronic cocaine response and addiction. Conversely, high-estrogenic females respond to cocaine by preferential chromatin closing, providing a mechanism for limiting cocaine-driven chromatin and synaptic plasticity. We find that physiological estrogen withdrawal, early-life stress, and absence of one X chromosome all nullify the protective effect of a high-estrogenic state on cocaine conditioning in females. Our findings offer a molecular framework to enable understanding of sex-specific neuronal mechanisms underlying cocaine use disorder.

Transient receptor potential M2 channel in the hypothalamic preoptic area and its impact on thermoregulation during menopause

BackgroundDecreased estrogen levels can cause abnormal thermosensitivity of the preoptic area (POA) in the hypothalamus during menopause, which may cause hot flashes. Thermosensitive transient receptors (ThermoTRPs) affect the thermosensitivity of neurons. It is worth exploring whether ThermoTRPs change under low estrogen state and participate in the abnormal thermoregulation of POA. MethodsAdult female Sprague–Dawley rats were randomly divided into sham operation (SHAM), ovariectomy (OVX) and estrogen treatment after ovariectomy (OVX+E) groups. Under 10 ℃, 18 ℃, 25 ℃, 37 ℃ and 45 ℃ incubations, their skin temperature was monitored and the expression of TRPA1, TRPM8, TRPM2, and TRPV1 in POA were investigated. ResultsThe skin temperature of ovariectomized rats changed faster and more dramatically under different incubation temperatures. The results at mRNA level show that only the expression of TRPM2 decreased in POA of OVX group compared with the other two groups at 25 ℃, TRPA1 expression in POA of the three groups increased at 10 ℃, TRPM8 increased at 10 ℃ and 18 ℃, TRPV1 increased at 10 ℃ and 45 ℃, while the expression of TRPM2 decreased at 10 ℃ and 18 ℃ and increased at 37 ℃ and 45 ℃. In all these cases, the magnitudes of the changes were less in the OVX group relative to the other two groups. The further immunohistochemical and Western blot results of TRPM2 and the activated TRPM2 positive cells labeled by c-Fos were consistent with the results of mRNA level. ConclusionsThe expression and thermosensitivity of TRPM2 in POA changed greatly under different incubation temperatures, but the changes in ovariectomized rats were less. This may be the key factor triggering thermoregulation dysfunction under low estrogen and may cause hot flashes.

Traditional and Emerging Sex-Specific Risk Factors for Cardiovascular Disease in Women.

Cardiovascular disease (CVD) remains a major health threat in women. While traditional CVD risk factors such as hypertension, hyperlipidemia, diabetes, and smoking have been recognized for over 50 years, optimal control of these risk factors remains a major challenge. Unique sex-specific risk factors such as adverse pregnancy outcomes, premature menopause and low estrogen states, and chronic autoimmune inflammatory disorders also contribute to increased CVD risk in women. In addition, psychological risk factors such as stress, depression, and social determinants of health may have a disproportionately adverse impact in women. An improved understanding of traditional and emerging sex-specific CVD risk factors and management of modifiable factors is critical for clinicians who provide care for women. Early recognition and treatment of risk factors may alter the trajectory of adverse CVD events. A multi-disciplinary approach with team-based care involving multiple specialists and improved, targeted educational efforts are needed to reduce CVD risk factors and its adverse consequences in women.

Identification of Risk Factors in the Development of Heterotopic Ossification After Primary Total Hip Arthroplasty.

PurposeHeterotopic ossification (HO) is a process by which bone forms abnormally in soft tissues. Known risk factors for developing HO include male sex, spinal cord injury, trauma, and surgery. We investigated additional risk factors in the development of HO after hip arthroplasty.MethodsWe performed a retrospective review of electronic medical records of 4070 individuals who underwent hip arthroplasty from September 2010 to October 2019 at the University of California, San Francisco Hospital. Demographics, anthropometrics, medications, and comorbid conditions were used in logistic regression analysis to identify factors associated with the development of HO.ResultsA total of 2541 patients underwent primary hip arthroplasty in the analyzed timeframe (46.04% men, mean age at procedure: 62.13 ± 13.29 years). The incidence of postsurgical HO was 3% (n = 80). A larger proportion of individuals who developed HO had underlying osteoporosis (P < 0.001), vitamin D deficiency (P < 0.001), spine disease (P < 0.001), type 1 or 2 diabetes (P < 0.001), amenorrhea (P = 0.037), postmenopausal status (P < 0.001), parathyroid disorders (P = 0.011), and history of pathologic fracture (P = 0.005). Significant predictors for HO development were Black/African American race [odds ratio (OR) 2.97, P = 0.005], preexisting osteoporosis (OR 2.72, P = 0.001), spine disease (OR 2.04, P = 0.036), and low estrogen states (OR 1.99, P = 0.025). In the overall group, 75.64% received perioperative nonsteroidal anti-inflammatory drugs (NSAIDs), which negatively correlated with HO formation (OR 0.39, P = 0.001).ConclusionsWe identified new factors potentially associated with an increased risk of developing HO after primary hip arthroplasty, including African American race, osteoporosis, and low estrogen states. These patients may benefit from HO prophylaxis, such as perioperative NSAIDs.

Cyclic estrogen and progesterone during instrumental acquisition contributes to habit formation in female rats

Habit formation is thought to involve two parallel processes that are mediated by distinct neural substates: one that suppresses goal-directed behavior, and one that facilitates stimulus-response (S-R) learning, which underscores habitual behavior. In previous studies we showed that habitual responding emerges early during instrumental training in gonadally-intact female, compared to male, rats. The present study aimed to determine the role of ovarian hormones during instrumental acquisition in the transition from goal-directed to habitual behavior in female rats. Ovariectomized (OVX) female rats were given subcutaneous silastic capsules that released low levels of 17-β estradiol (E2) to maintain estrogen receptor availability. Rats were assigned to one of three hormone treatment conditions: no additional hormone replacement (Control group), replacement with high E2 (High E2 group), or replacement with high E2 followed by progesterone (High E2 + P4 group). Hormone replacement occurred twice during acquisition to mimic natural hormone fluctuations. At test, the Control and High E2 groups demonstrated responding that was sensitive to devaluation by lithium chloride-induced illness, indicating goal-directed behavior. In contrast, the High E2 + P4 group exhibited a pattern of devaluation-insensitive, habitual responding, that suggested the suppression of goal-directed processes. In a follow-up experiment, similar procedures were conducted, however during acquisition, OVX rats were given cyclic high E2 plus medroxy-progesterone (MPA), a form of progesterone that does not metabolize to neuroactive metabolites. In this group, goal-directed behavior was observed. These data indicate that habit formation is not facilitated in low estrogen states, nor in the presence of cyclic high E2. However, cyclic high E2, together with progesterone during acquisition, appears to facilitate the early emergence of habitual responding. Furthermore, these data suggest that a neuroactive progesterone metabolite, like allopregnanolone, in combination with high cyclic E2, supports this phenomenon.

Acute Delta 9-tetrahydrocannabinol administration differentially alters the hippocampal opioid system in adult female and male rats.

Our prior studies demonstrated that the rat hippocampal opioid system can undergo sex-specific adaptations to external stimuli that can influence opioid-associated learning processes. This opioid system extensively overlaps with the cannabinoid system. Moreover, acute administration of Δ9 Tetrahydrocannabinoid (THC), the primary psychoactive constituent of cannabis, can alter cognitive behaviors that involve the hippocampus. Here, we use light and electron microscopic immunocytochemical methods to examine the effects of acute THC (5mg/kg, i.p., 1h) on mossy fiber Leu-Enkephalin (LEnk) levels and the distribution and phosphorylation levels of delta and mu opioid receptors (DORs and MORs, respectively) in CA3 pyramidal cells and parvalbumin dentate hilar interneurons of adult female and male Sprague-Dawley rats. In females with elevated estrogen states (proestrus/estrus stage), acute THC altered the opioid system so that it resembled that seen in vehicle-injected females with low estrogen states (diestrus) and males: (1) mossy fiber LEnk levels in CA2/3a decreased; (2) phosphorylated-DOR levels in CA2/3a pyramidal cells increased; and (3) phosphorylated-MOR levels increased in most CA3b laminae. In males, acute THC resulted in the internalization of MORs in parvalbumin-containing interneuron dendrites which would decrease disinhibition of granule cells. In both sexes, acute THC redistributed DORs to the near plasma membrane of CA3 pyramidal cell dendrites, however, the dendritic region varied with sex. Additionally, acute THC also resulted in a sex-specific redistribution of DORs within CA3 pyramidal cell dendrites which could differentially promote synaptic plasticity and/or opioid-associated learning processes in both females and males.

Incidence and Treatment Efficacy of Trigger Finger in the Breast Cancer Population on Aromatase Inhibitors.

Trigger finger is a common hand complaint of the general population. Limited literature exists implicating a low-estrogen state in patients on aromatase inhibitor (AI) therapy for breast cancer who develop trigger finger. The authors' objective was to determine the incidence and treatment outcomes of this population. A single-center retrospective chart review was conducted on patients with a diagnosis of breast cancer on AI who developed trigger finger from 2010 to 2019. The total population of patients during this time served as our population, and patients with breast cancer not on AI with trigger finger served as our control. Primary outcomes included total number of injections and need for surgery. Secondary outcomes included risk factors for surgery. χ2 analysis and logistical regression model determined the significance of primary and secondary outcomes, respectively. In all, 192 patients of a population size of 664 751 met our study group criteria. The study group showed a higher incidence of trigger finger (5.1% vs 1.3%; P < .001) compared with our population. Patients treated with AI for breast cancer had both higher incidence of trigger finger (5.1% vs 1.5%, P < .001) and injections (77.1% vs 66.5%, P < .001) compared with patients not on AI therapy. Independent risk factors requiring surgical treatment were found in patients with diabetes (odds ratio [OR], 3.54; P = .01) and in patients with concomitant radiation therapy (OR, 3.17; P = .02). This study demonstrates for the first time the incidence, treatment outcomes, and surgical risk factors of trigger finger in patients on AI therapy for breast cancer.

Abstract 029: Menopause Alters The Gut Microbiome In Hispanic/Latina Women Of The Hispanic Community Health Study/Study Of Latinos (HCHS/SOL), With Implications For Metabolic Syndrome

Introduction: During menopause, women experience an increase in cardiometabolic risk factors, thought to relate to aging and loss of endogenous estrogen. Menopause may also influence the gut microbiome, which plays a role in cardiometabolic risk. However, the menopause-microbiome relationship has not been examined in a large study and implications for disease are unknown. Hypothesis: Menopause alters gut microbiome composition, which may affect post-menopausal disease risk. Methods: Menopause was defined by self report of 12 months of amenorrhea, not due to surgery or other non-natural causes. Shotgun sequencing was used on stool from 2370 participants (301 pre-menopausal women; 1072 post-menopausal women; 997 men). Between pre- and post-menopausal women and age-matched men, we compared microbiome diversity, composition, and taxa. Results: Pre-menopausal women had higher gut microbiome diversity than post-menopausal women and men (all p&lt;0.05, Figure 1a). Post-menopausal women also differed in overall microbiome composition from pre-menopausal women (p=0.005), while older and younger men did not differ from each other (p=0.70) (Figure 1b). We identified differentially abundant taxa between post- and pre-menopausal women, including class Betaproteobacteria and its genus Sutterella (enriched in post-menopause), and genera Shigella and Escherichia (depleted in post-menopause); associations were similar in comparisons of men vs. women, though the taxa did not differ between older and younger men (Figure 1c). In post-menopausal women, higher abundance of Shigella and Escherichia was associated with impaired fasting glucose (≥100 mg/dL or diabetes treatment) (Figure 1d). Conclusions: Menopause is associated with a gut microbiome more similar to that of men, perhaps related to the common condition of a low estrogen state. In HCHS/SOL, menopause was associated with depletion of gut pathogens, which were related to increased diabetes risk. These findings require confirmation in other racial/ethnic groups.

Maintained Bone Density in Young Hypoestrogenized Women with a High BMI: Case Series.

Estrogen deficiency and obesity are factors that affect bone mass in a manner that is independent and in opposing directions. Obesity favours higher bone mass and increased bone formation whereas estrogen deficiency leads to significant bone loss in leaner individuals. To report the impact of the competing effects of a hypoestrogenized state and obesity on long-term bone health, we present two cases of young chronically hypoestrogenized females whose bone parameters were assessed with high-resolution peripheral quantitative computed tomography (HR-pQCT) and revealed a bone mineral density and microstructure that did not change despite the long history of a low estrogen state. As evidenced by the outcomes for these patients, the obesity-related effect on bone mass may be dominant when obesity is marked and appears to be highly protective even in the setting of sub-physiologic circulating estrogen. Recognition of this interaction should be considered in decisions around estrogen replacement therapy in such cases.

11. Lichen Sclerosus Throughout Childhood and Adolescence: Not Only a Premenarchal Disease

<h3>Background</h3> Vulvar lichen sclerosus is a benign dermatologic condition marked by inflammation and epithelial thinning. It is thought to mostly occur in low estrogen states, though a few studies have shown that symptoms may persist following puberty. The purpose of this study is to characterize a cohort of pediatric and adolescent females with lichen sclerosus and compare age of symptom onset, documented symptoms and duration, and exam findings. <h3>Methods</h3> This is an IRB exempt retrospective cohort study of females aged 0-21 years seen between 2000 and 2020 with the diagnosis of lichen sclerosus by ICD-9 or 10 code. Study variables included age at symptom onset, age at diagnosis, presenting symptoms, exam findings, and presence of symptoms before and after menarche. Groups were compared using the chi square test of independence and Fisher's exact test. Data were analyzed using SPSS Version 27. <h3>Results</h3> One hundred patients met inclusion criteria. Forty patients noted presence of symptoms following menarche; 26 (65%) reported first symptom onset after age 13, while 14 (35%) had symptoms that started in childhood and persisted after menarche. Eleven patients with childhood onset of symptoms had no documented report of symptoms following menarche. Of the remaining 49 patients, 25 are still under age 13 and 24 were lost to follow-up. After excluding the 7 patients with reported childhood onset of symptoms that were not seen until adolescence, the patients with onset of symptoms at age 13 or older (n=26) were less likely to present with vaginal bleeding (3.8% vs 25.4%, p=0.02), dysuria (15.4% vs 32.8%, p=0.09), and constipation or painful defecation (0% vs 14.9%, p=0.05) than the younger cohort (n=67). Vulvar itching (61.5% vs 71.6%) and pain (38.5% vs 44.8%) were similar in both groups. Seven patients (26.9%), ranging in age from 17-21 years, presented with dyspareunia. On initial physical exam, the patients with symptom onset at age 13 or older were more likely to have clitoral adhesions (19.2% vs 3%, p=0.02), loss of labia minora (38.5% vs 3.0%, p<0.01), and loss of interlabial sulcus (11.5% vs 1.5%, p=0.06) than the younger cohort. They were less likely to have subepithelial hemorrhage (3.8% vs 23.9%, p=0.03). <h3>Conclusions</h3> This study suggests that lichen sclerosus can present or persist after menarche in at least 40% of patients. It can cause architectural vulvar changes that interfere with sexual function, which may be the presenting complaint. It is important for providers to consider lichen sclerosus as a new diagnosis in both pre and postmenarchal patients as well as monitor for progression of disease throughout the pubertal transition and adolescence.

Vulvodynia.

Vulvodynia is a heterogenous, chronic pain condition of unknown etiology that affects 7% to 15% of women. It affects sexual function and quality of life. Vulvodynia can be primary or secondary, localized or generalized, and spontaneous or provoked. Contributing factors for provoked vulvodynia might include vulvovaginal infections, low estrogen states, and underlying anxiety disorder. Generalized vulvodynia likely arises from underlying connective tissue or neurological dysfunction. Vulvodynia treatment must be individualized on the basis of the patient's presentation and physical examination findings. Surgical excision of the vulvar vestibule has high success rates but other modalities showing success include pelvic floor physical therapy and cognitive-behavioral therapy.

Effects of resveratrol on bone metabolism and bone turnover related indexes in ovariectomized osteoporosis rats

This study aimed to investigate the effects of resveratrol (RES) on bone metabolism and bone turnover related indexes in ovariectomized osteoporosis rats. 48 clean grade adult healthy unmated female SD rats were randomly divided into 6 groups, including normal control group (NCG), osteoporosis model group (OP MG), estrogen treatment group (17β-E2 group), RES low dose group (RES-L), RES medium-dose group (RES-M) and RES high dose group (RES-H). The rats in NCG and OP MG were given distilled water once a day and the rats in the other two groups were given 17β-E2 and resveratrol respectively. The levels of serum calcium (S-Ca), serum phosphorus (S-P), urinary calcium (U-Ca/Cr) and urinary phosphorus (U-P/Cr) were measured with an automatic biochemistry analyzer. The levels of serum osteocalcin (BGP), alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP), type I amino front-end peptide (PINP), type I collagen strong carboxyl peptide (CTX-I), urine deoxypyridinoline (DPD) and serum estrogen were detected by enzyme-linked immunosorbent assay (ELISA). In the OP group, the serum estrogen levels, S-Ca and S-P decreased significantly and the expression of U-Ca/Cr and U-P/Cr increased significantly (P&lt; 0.05). Compared with the OP group, the expression of S-Ca and S-P increased significantly and the expression of U-Ca/Cr and U-P/Cr decreased significantly (p&lt; 0.05) after treatment. The levels of TRAP, BGP, DPD and CTX-I in the OP group increased significantly (P&lt; 0.05). Compared with the OP group, the levels of TRAP decreased significantly (P&lt; 0.05). The levels of PINP and ALP in OP MG increased significantly (P&lt; 0.05). IP and ALP increased in the middle and lower levels (P&lt; 0.05). The bone mineral density of the OP group decreased significantly (P&lt; 0.05). Resveratrol can affect the changes in bone turnover in ovariectomized rats, promote bone formation in low estrogen state and inhibit bone resorption. Resveratrol may have a protective effect on the bone of ovariectomized rats.

Estrogen receptor inhibition enhances cold-induced adipocyte beiging and glucose tolerance.

Background Low estrogen states, exemplified by postmenopausal women, are associated with increased adiposity and metabolic dysfunction. We recently reported a paradox, in which a conditional estrogen receptor-alpha (ERα) mutant mouse shows a hyper-metabolic phenotype with enhanced brown/beige cell formation (“browning/beiging”).Hypothesis These observations led us to consider that although systemic deficiency of estrogen or ERα in mice results in obesity and glucose intolerance at room temperature, cold exposure might induce enhanced browning/beiging and improve glucose metabolism.Methods and results Remarkably, studying cold-exposure in mouse models of inhibited estrogen signaling - ERαKO mice, ovariectomy, and treatment with the ERα antagonist Fulvestrant - supported this notion. ERα/estrogen-deficient mice demonstrated enhanced cold-induced beiging, reduced adiposity and improved glucose tolerance. Fulvestrant was also effective in diet-induced obesity settings. Mechanistically, ERα inhibition sensitized cell-autonomous beige cell differentiation and stimulation, including β3-adrenoreceptor-dependent adipocyte beiging.Conclusion Taken together, our findings highlight a therapeutic potential for obese/diabetic postmenopausal patients; cold exposure is therefore predicted to metabolically benefit those patients.

Sex and Estrous Cycle Effects on Anxiety- and Depression-Related Phenotypes in a Two-Hit Developmental Stress Model.

Stress during sensitive developmental periods can adversely affect physical and psychological development and contribute to later-life mental disorders. In particular, adverse experiences during childhood dramatically increase the risk for the development of depression and anxiety disorders. Although women of reproductive age are twice as likely to develop anxiety and depression than men of the corresponding age, little is known about sex-specific factors that promote or protect against the development of psychopathology. To examine potential developmental mechanisms driving sex disparity in risk for anxiety and depression, we established a two-hit developmental stress model including maternal separation in early life followed by social isolation in adolescence. Our study shows complex interactions between early-life and adolescent stress, between stress and sex, and between stress and female estrogen status in shaping behavioral phenotypes of adult animals. In general, increased locomotor activity and body weight reduction were the only two phenotypes where two stressors showed synergistic activity. In terms of anxiety- and depression-related phenotypes, single exposure to early-life stress had the most significant impact and was female-specific. We show that early-life stress disrupts the protective role of estrogen in females, and promotes female vulnerability to anxiety- and depression-related phenotypes associated with the low-estrogenic state. We found plausible transcriptional and epigenetic alterations in psychiatric risk genes, Nr3c1 and Cacna1c, that likely contributed to the stress-induced behavioral effects. In addition, two general transcriptional regulators, Egr1 and Dnmt1, were found to be dysregulated in maternally-separated females and in animals exposed to both stressors, respectively, providing insights into possible transcriptional mechanisms that underlie behavioral phenotypes. Our findings provide a novel insight into developmental risk factors and biological mechanisms driving sex differences in depression and anxiety disorders, facilitating the search for more effective, sex-specific treatments for these disorders.

Bariatric Surgery: Weighing In on Bone Loss.

Bariatric Surgery: Weighing In on Bone Loss.

Impact of Breastfeeding on Lactating Women Bone Health: A Survey based Study in Northern Region of Saudi Arabia

Bone fitness is considered as an important public health problem especially in post-menopausal women. Numerous studies have investigated to understand beneficiary effects of mother milk on the baby but very few studies have been conducted to understand the effect of breastfeeding on the bone health of lactating women. We hypothesized that high calcium demand during pregnancy and lactation and low estrogenic state support may affect on the bone health immediately after child birth and later in the life. Based upon this theory we have conducted a survey based study in the Northern region of Saudi Arabia to assess the health status of the lactating mothers. Total 376 lactating mothers were interviewed during the period of 16 Nov 2016 to 23 March 2017. A pre-tested and structured questionnaire was provided at their homes to collect information. The result revealed that among 376 participants, 7% complaining about generalized pain, 41% suffering from joint pain, 24%, suffering from lower back pain and 1% of mothers suffered from fracture. This survey highlights important but ignored aspect of the bone health of lactating women in the Northern region of Saudi Arabia. During pregnancy, women consume calcium supplements to satisfy their requirement of developing baby but after child birth, they stop calcium supplement though it's needed. Based upon this study we state that there is a need to prepare and follow guidelines for the use of calcium supplement for the lactating women to maintain bone health.

Abstract P3-04-01: ESR1 mutations, ESR1 fusions and co-occurring alterations assessed in breast cancer tumors

Abstract Background: ESR1 mutations and fusions arise in hormone receptor positive (ER+ and/or PR+) breast cancer (HR+ BC) patients after aromatase inhibitor (AI) therapy (low estrogen states), to become constitutively active in a ligand-independent manner. Patients with ESR1 mutations exhibit worse prognosis and outcome with no particular benefit of chemotherapy vs tamoxifen treatment (Augusto, et al. ASCO 2016). A retrospective analysis of the ESR1 mutation frequency and co-occurring alterations that could guide subsequent therapy approaches was investigated. Methods: Molecular profiles of 416 breast cancer patients [HR+ (n=237), HER2+ (n=29) and TNBC (n=139)] were assessed. Protein expression (IHC) and gene amplification (ISH) were performed. Genomic testing included 592-gene hybrid-capture NGS [NextSeq Illumina platforms] and ArcherDx fusion assay based on anchored multiplex PCR (AMP) FusionPlex Solid Tumor. ESR1 variant (ESR1var) (mutation/fusion) profiles and HR+ BC patients lacking genomic ESR1 alterations (ESR1 WT) were compared; Pearson's chi-squared test was used to test for significant differences. Results: An ESR1 mutation (point mutations, insertion-deletions [n=49] and fusions [n=4]) was detected in 13% (53 /416) of the specimens, and this constitutes 21% (50/237) of all HR+ breast cancers. Two TNBC patients exhibited ESR1 variants (H398Y in exon 7 and A491S in exon 9, both are classified as variants of unknown significance). ESR1 mutations were not detected in HER2+ BC. Seventy-seven percent of patients with ESR1 mutations were detected in metastatic specimens (p=0.03), with liver (19/53 or 36%) and bone (8/53 or 15%) specimens as the most frequent sources for ESR1 variant (ESR1var) positivity. The most common alleles detected were: D538G (24%), Y537S (18%), E380Q (14%) and L536H (4%); 15 other additional alleles were detected (each 2%). ESR1 fusions were detected in 4 ESR1 WT patients: ESR1-ATP2B2, ESR1-MKL1/ESR1-TNRC6B, ESR1-ARNT2 and ESR1-C6ORRF211. We next compared ESR1var (mutation/fusion) profiles to HR+ breast patients lacking genomic ESR1 alterations (ESR1 WT). ER expression was present in 100% and 96% of ESR1var and ESR1 WT BC, respectively, however expression of PR was negative in 22% and 38% of ESR1var and WT BC, respectively (p=0.05). Significantly higher rates of other gene amplification events observed in ESR1var vs. ESR1 WT BC included: c11orf30 [EMSY, BRCA2 interacting transcriptional repressor] (20% vs. 7%), CCND1 (51% vs.28%), CCND2 (6% vs. 0%), FGF3 (37% vs. 16%), FGF4 (40% vs. 12%) and FGF19 (40% vs. 15%), whereas cMYC was more frequently amplified in ESR1 WT BC (17% vs. 2%); all p-values &amp;lt;0.05. KRAS mutations were higher in ESR1var vs WT BC (4% vs. 0%; p=.004). Alterations in the PIK3CA pathway were common in both ESR1var and ESR1 WT BC: mutations in PIK3CA, AKT and PTEN were observed in 27% and 26%; 4% and 7%; 6% and 5%, respectively, and PTEN loss by IHC in 29% and 26%. Conclusions: ESR1 mutations and fusions are detected in 21% of HR+ BC, the majority of which are in metastatic sites. Amplifications of genes involved in downstream regulatory pathways were present and may contribute to the poor prognosis of ESR1var HR+ BC. Correlation with antecedent therapy is currently underway. Citation Format: Tan A, Feldman R, Pohlmann P, Lynce F, Swain S, Nunes MR, Gatalica Z, Ghazalpour A, Swensen J, Vidal G, Obeid E, Vacirca J, Isaacs C, Schwartzberg L. ESR1 mutations, ESR1 fusions and co-occurring alterations assessed in breast cancer tumors [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P3-04-01.