Background:Despite significant progress in treating estrogen receptor (ER)-positive breast cancer (BC), the recurrence risk remains high and mechanisms driving initial ductal tumorigenesis is unclear. Given an emerging role of LINE-1 (L1), HERV-K and other endogenous retroviral elements (EREs) in cancer, we tested the hypothesis that their expression can be induced by estrogen and blocked by treatment with lamivudine (3TC), an HIV-1 drug that can also inhibit L1 and HERV-K encoded reverse transcriptases (RTs) and ERE propagation. Methods:This is a preclinical study using the well-established ACI rat model of spontaneous estrogen-driven BC to examine the earliest stages of cancer development and the effects of 3TC. Rat mammary glands and peripheral blood mononuclear cells (PBMCs) were analyzed by immunohistochemistry (IHC) and RNA sequencing (RNAseq). A panel of human BC cell lines and tissue microarrays (TMAs) were assessed by RT-qPCR, immunoblotting and IHC to evaluate ERE RNA and protein expression and a potential association with the ER- and FOXA1- positive BC. Results:Development of estrogen-induced ER+ and FOXA1+ pre-invasive ductal tumors in rat mammary glands was accompanied by systemic inflammation and upregulation of ERE RNAs in mammary glands and PBMCs; these effects were attenuated by oral administration of 3TC. L1 and HERV RNAs and proteins were also detected in human BC cell lines and in tumor tissues with high ER and FOXA1 protein levels, suggesting that their expression is an intrinsic part of the ER- and FOXA1- driven oncogenic programs. Conclusion:Using a rat model, our study demonstrates estrogen dependent ERE expression in pre-invasive ER+FOXA1+ mammary tumor cells and in circulating immune cells, activation of innate immune responses, and the inhibitory effects of 3TC. ERE RNAs and proteins are also highly expressed in human ER+FOXA1+ breast cancers, warranting further investigation into their functions and a potential use of 3TC as a preventive strategy for reducing a risk of cancer progression, especially in woman with ER+FOXA1+ tumors.

Decidualization triggered by human endometrial stromal cells (HESCs) is a critical step in the establishment of endometrial receptivity, which provides an ideal environment for embryo implantation. However, the specific regulatory networks and core genes involved in the decidualization process remain incompletely characterized. Here, we identified ubiquitin-conjugating enzyme 2C (UBE2C), a key cell cycle regulator, as a pivotal factor in decidualization through comprehensive in vitro and in vivo investigations. RNA sequencing revealed UBE2C as a potential hub gene in the human endometrium. We found UBE2C expression was significantly downregulated in secretory-phase endometrium and in vitro decidual HESCs. Functional studies demonstrated that UBE2C overexpression attenuated decidual marker expression and disrupted normal proliferation in HESCs, mechanistically linked to NF-κB signaling pathway. Consistent with in vitro findings, mice studies showed UBE2C downregulation in peri-implantation uterine tissue, with uterine-specific UBE2C overexpression impairing both embryo implantation and decidualization. Further mechanistic exploration identified transcription factor AP-2 alpha (TFAP2A) as a novel mediator of UBE2C. Decidual stimulation promoted TFAP2A degradation via the ubiquitination pathway. Clinical relevance was established through the observation of elevated UBE2C and TFAP2A expression in secretory endometrium from recurrent implantation failure (RIF) patients. In conclusion, for endometrial decidualization and embryo implantation, TFAP2A-mediated downregulation of UBE2C is required. Dysregulation of this axis may compromise endometrial receptivity, suggesting UBE2C and TFAP2A as promising therapeutic targets for decidualization-related disorders such as RIF.

Aberrant hyperactivation of the Hippo pathway effector YAP/TEAD complex causes tissue overgrowth and tumorigenesis in various cancers, including endometrial cancer (EC). The transcription factor SOX17 (SRY [sex-determining region Y]-box 17) is frequently mutated in EC; however, SOX17 mutations are rare in other cancer types. The molecular mechanisms underlying SOX17 mutation–induced EC tumorigenesis remain poorly understood. Here, we demonstrate that SOX17 serves as a tumor suppressor to restrict the proliferation, migration, invasion, and anchorage-independent growth of EC cells, partly by suppressing the transcriptional outputs of the Hippo-YAP/TEAD pathway. SOX17 binds to TEAD transcription factors through its HMG domain and attenuates the DNA-binding ability of TEAD. SOX17 loss by inactivating mutations leads to the malignant transformation of EC cells, which can be reversed by small-molecule inhibitors of YAP/TEAD or cabozantinib, an FDA-approved drug targeting the YAP/TEAD transcriptional target AXL. Our findings reveal novel molecular mechanisms underlying Hippo-YAP/TEAD pathway–driven EC tumorigenesis, and suggest potential therapeutic strategies targeting the Hippo-YAP/TEAD pathway in SOX17-mutated EC.

Chronic salpingitis is one of the most common causes of female infertility. Luteal support is a critical step for embryo transfer. Here, we evaluated the effects of two luteal support regimens, intramuscular progesterone (IMP) and progesterone vaginal gel (VAG), on the pregnancy outcomes in patients with chronic salpingitis undergoing vitrified-warmed embryo transfer. This study retrospectively analyzed 2240 patients with chronic salpingitis undergoing vitrified-warmed embryo transfer from 2015 to 2022 at our center. Patients were categorized into IMP group (n = 1039) and VAG group (n = 1201). Inverse probability of treatment weighting (IPTW) was used to balance baseline characteristics. Univariate and multivariate logistic regression models were conducted to analyze pregnancy outcomes. After IPTW, baseline demographic characteristics were balanced and outcome indicators were comparable. Crude analysis showed a higher live birth rate (OR 1.25, 95

Cervical human papillomavirus (HPV) infection is believed to increase the risks of pregnancy failure and abortion, however, whether the uterine cavity HPV infection reduces pregnancy rate or increases miscarriage rate remains unclarified in infertile women undergoing assisted reproductive technology (ART) treatment. Therefore, we aimed to assess ART outcomes in the presence of intrauterine HPV. This was a hospital-based multicenter (five reproductive medicine centers) matched cohort study. This study involved 4153 infertile women undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection treatment in five reproductive medicine centers between October 2018 and 2020. The spent embryo transfer media sample with endometrium tissue were collected and performed with flow-through hybridization and gene chips to detect HPV DNA. According to basic characteristics, HPV-positive and negative patients were matched in a ratio of 1:4 by age, body mass index transfer timing, transfer type, and number of embryos transferred. The primary outcome was pregnancy and clinical miscarriage rates in the transfer cycle underwent HPV detection. 92 HPV-positive and 368 HPV-negative patients were screened and analyzed statistically. Univariate analysis showed uterine cavity HPV infection resulted in lower rates of ongoing pregnancy (31.5% vs. 44.6%; p=0.023), implantation (32.3% vs. 43.1%; p=0.026), biochemical pregnancy (47.8% vs. 62.5%; p=0.010), and clinical pregnancy (40.2% vs. 54.3%; p=0.015) compared with HPV negative group. The infertile female with positive HPV also had a slightly higher frequency of biochemical miscarriage (15.9% vs. 13.0%; p=0.610) and clinical miscarriage (24.3% vs. 15.5%; p=0.188). These findings suggest that HPV infection in the uterine cavity is a high risk for ART failure. HPV screening is recommended before ART treatment, which may be benefit to improving pregnancy outcome.

An effective tool to assess embryo quality in the assisted reproduction clinical practice will enhance successful implantation rates and mitigate high risks of multiple pregnancies. Potential biomarkers secreted into culture medium (CM) during embryo development enable rapid and noninvasive methods of assessing embryo quality. However, small volumes, low biomolecule concentrations, and impurity interference collectively preclude the identification of quality-related biomarkers in single blastocyst CM. Here, we developed a noninvasive trace multiomics approach to screen for potential markers in individual human blastocyst CM. We collected 84 CM samples and divided them into high-quality (HQ) and low-quality (LQ) groups. We evaluated the differentially expressed proteins (DEPs) and metabolites (DEMs) in HQ and LQ CM. A total of 504 proteins and 189 metabolites were detected in individual blastocyst CM. Moreover, 9 DEPs and 32 DEMs were identified in different quality embryo CM. We also categorized HQ embryos into positive implantation (PI) and negative implantation (NI) groups based on ultrasound findings on day 28. We identified 41 DEPs and 4 DEMs associated with clinical implantation outcomes in morphologically HQ embryos using a multiomics analysis approach. This study provides a noninvasive multiomics analysis technique and identifies potential biomarkers for clinical embryo developmental quality assessment.

The progestin regimen is one of the main therapeutic strategies for women with endometrial cancer who undergo conservative management. Although many patients respond well to initial therapy, progestin-refractory disease inevitably emerges, and the molecular basis underlying progestin resistance has not been comprehensively elucidated. Herein, they demonstrated progestin results in p38-dependent IDH1 Thr 77 phosphorylation (pT77-IDH1). pT77-IDH1 translocates into the nucleus and is recruited to chromatin through its interaction with OCT6. IDH1-produced α-ketoglutarate (αKG) then facilitates the activity of OCT6 to promote focal adhesion related target gene transcription to confer progestin resistance. Pharmacological inhibition of p38 or focal adhesion signaling sensitizes endometrial cancer cells to progestin in vivo. The study reveals p38-dependent pT77-IDH1 as a key mediator of progestin resistance and a promising target for improving the efficacy of progestin therapy.

Background The enriched proteins within in vitro fertilisation (IVF)-generated human embryonic microenvironment could reverse progestin resistance in endometrial cancer (EC). Methods The expression of thymic stromal lymphopoietin (TSLP) in EC was evaluated by immunoblot and IHC analysis. Transcriptome sequencing screened out the downstream pathway regulated by TSLP. The role of TSLP, androgen receptor (AR) and KANK1 in regulating the sensitivity of EC to progestin was verified through a series of in vitro and in vivo experiments. Results TSLP facilitates the formation of a BMP4/BMP7 heterodimer, resulting in activation of Smad5, augmenting AR signalling. AR in turn sensitises EC cells to progestin via KANK1. Downregulation of TSLP, loss of AR and KANK1 in EC patients are associated with tumour malignant progress. Moreover, exogenous TSLP could rescue the anti-tumour effect of progestin on mouse in vivo xenograft tumour. Conclusions Our findings suggest that TSLP enhances the sensitivity of EC to progestin through the BMP4/Smad5/AR/KANK1 axis, and provide a link between embryo development and cancer progress, paving the way for the establishment of novel strategy overcoming progestin resistance using embryo original factors.

1. Basic characteristics of endometrial hyperplasia and EC patients. Supplementary Table 2. Basic information of clinical samples for Immunohistochemistry. Supplementary Table 3. Primers used.

Induction of CD68+CD163+ macrophages from THP-1 cells

目的:研究KANK1在子宫内膜癌中的表达以及对子宫内膜癌细胞增殖及迁移的影响.方法:(1)TCGA数据库分析KANK1在子宫内膜癌中的表达和生存期分析.(2)采用实时荧光定量聚合酶链反应验证转染KANK1质粒的效果.采用Ishikawa和ECC1这两种子宫内膜癌细胞来探讨KANK1对子宫内膜癌的细胞周期和凋亡的影响.通过Western blot检测细胞周期相关蛋白的表达,以及流式细胞术检测细胞周期和凋亡水平.(3)通过Transwell小室实验和划痕实验检测细胞的侵袭和转移能力.结果:TCGA数据库分析发现KANK1在子宫内膜癌中低表达且与患者预后良好相关.过表达KANK1下调了 Cyclin D1和Cyclin D2的蛋白水平,并将细胞周期阻滞在G1期.流式细胞术检测发现过表达KANK1组的细胞凋亡水平(Ishikawa:22.7％;ECC1:19.0％)比对照组(Ishikawa:18.1％;ECC1:15.3％)高,差异具有统计学意义.Transwell迁移和侵袭实验结果表明过表达KANK1组的子宫内膜癌细胞侵袭和转移能力减弱.结论:本研究证明了 KANK1在子宫内膜癌中发挥抑癌作用.KANK1高表达与子宫内膜癌的预后良好成正相关.KANK1通过抑制癌细胞周期和促进肿瘤细胞凋亡发挥抑制子宫内膜癌增殖的作用.此外,KANK1抑制了子宫内膜癌的侵袭和转移.

IL-17 receptor antibody compromises the effect of IL-17A on ERα transcription up-regulation

The expression pattern of ERα in different cell lines

Serum estradiol levels were not elevated in patients with endometrioid adenocarcinoma after stratification by pathologic grading

Semi-quantitative evaluation of the immunohistochemical staining of IL-17A, TET1, 5-hmC, and ERα expression in endometrial hyperplasia and cancer lesions

Supplementary Materials and Methods and Supplementay Figure legends

IL-17A or overexpression of ERα promoted estradiol-driven endometrial cancer cell proliferation

Objective This study aimed to examine the efficacy of HRT with gonadotropin-releasing hormone agonist (GnRH-a) pre-treatment in women with male-factor infertility who underwent a frozen embryo transfer (FET) programme. Design Between January 2016 and October 2020, 2733 women with male-factor infertility who underwent the HRT protocol as the endometrial preparation method were enrolled at two Reproductive Medicine Centres. Patients were divided into two groups based on whether they had GnRH-a pre-treatment before HRTs: the GnRHa-HRT group and the HRT group. The inverse probability of treatment weighting (IPTW) method was conducted to balance patient baseline characteristics between treatment cohorts to reduce selection bias. The live birth rate was considered regarded as the primary pregnancy outcome. Results Multivariate logistic regression adjusted for confounding factors, the GnRHa-HRT group showed a notably higher rate of live birth (OR 2.154, 95% CI 1.636~2.835, P<0.001) when compared to the HRT group. Additionally, the rate of miscarriage was significantly lower in the GnRHa-HRT group. The GnRHa-HRT group had significantly higher rates of biochemical pregnancy, clinical pregnancy, multiple pregnancy, and term birth. Conclusion The endometrial preparation protocol of HRT with GnRH-a pre-treatment could obviously increase the live birth rate for women with male-factor infertility undergoing the FET programme.

AbstractBackgroundHigh‐risk human papillomavirus (hrHPV) infection is a key factor that alters cervicovaginal microbiota patterns and causes cervical intraepithelial neoplasias (CINs) or even cervical cancer. Although local excisional treatment can clear hrHPV infection and restore the cervicovaginal microbiota, it is unclear which cervicovaginal microbiota represents recovery. Our objective was to describe the cervicovaginal microbiota before and after treatments and to assess the association between the microbiota and HPV persistence.ResultsA cohort of 91 participants was classified into four groups (healthy control women and HPV16‐infected women with CIN I, CIN II/III, and squamous cell carcinoma [SCC]). Endocervical swabs were collected 3 months prior to treatment and at 3 months post‐treatment for bacterial 16S rRNA gene pyrosequencing and for HPV DNA testing. There was an increase in the number of Lactobacillus bacterial species present after the clinical treatments, and the community state type (CST) profiles were shifted from dysbiotic CSTs II and IV to Lactobacillus‐dominated CSTs I and III. Specifically, the composition of Geobacter and Prevotella before treatment and Lactobacillus secaliphilus after treatment might have been related to CIN I, the composition of Burkholderia before treatment and Lactobacillus iners after treatment might have been related to CIN II/III, and the composition of Atopobium and Aerococcus before treatment and Bacilli after treatment might have been related to SCC. Further functional predictions revealed that the composition differences were linked to infectious disease‐ and cancer‐related genes.ConclusionOur study provides an illustration of the changes in CSTs and the cervicovaginal microbiota before and after HPV16 clearance in each disease state.

RESEARCH QUESTION:What is the effect of letrozole use in patients undergoing frozen embryo transfer (FET) with normal ovulation? Although the number of FETs is increasing, an optimal protocol for FET (particularly vitrified-warmed embryo transfer) is yet to be determined. The aim of this study was to evaluate letrozole use on patients with normal menstrual cycles compared with hormone replacement therapy (HRT) cycles and natural cycles. DESIGN:The study involved 2849 patients. Patients were divided into three groups: HRT cycle (n = 2115), letrozole cycle (n = 532) and natural cycle (n = 202). Inverse probability of treatment weighting aimed to equate each group according to measured baseline covariates to achieve a comparison with reduced selection bias and live birth rate as main pregnancy outcome was analysed. RESULTS:In the crude analysis, the letrozole group had a higher live birth rate compared with the HRT cycle (OR 1.18, 95% CI 1.06 to 1.33) and natural cycle (OR 1.24, 95% CI 1.11 to 1.41); after adjusting for confounding factors, live birth rate was consistently higher in the letrozole group. Moreover, the biochemical pregnancy, clinical pregnancy, ongoing pregnancy and full-term delivery rates were higher in the letrozole group. CONCLUSION:For infertile women with normal menstrual cycle undergoing FET, mildly stimulated cycles with letrozole present a relatively large advantage compared with HRT cycle and natural cycle, with higher live birth pregnancy, indicating that letrozole administration could improve pregnancy outcomes in this population.