High Estrogen Receptor Expression Research Articles

Abstract PO2-26-01: Prognostic Role of HER2 Expression in Patients with ER-positive/HER2-negative Breast Cancer. Results from a Population-Based Cancer Registry Study

Abstract Background: Estrogen Receptor (ER)-positive (+)/Human Epidermal Growth Factor Receptor 2 (HER2)-negative (-) breast cancers (BCs) express variable protein levels of HER2, which can influence prognosis. Materials and Methods: This cohort study was conducted using data on all consecutive patients (pts) diagnosed with BC between 2005 and 2017, which were systematically and prospectively collected by the Emilia-Romagna Cancer Registry (Provinces of Piacenza, Parma, Modena, and Ferrara), Italy. The study included 13527 pts with ER+/HER2- BC. Tumors were classified by HER2 IHC score (0 [n=7155], 1+ [n=5186], or 2+ with negative FISH [n=1186]). Comparisons of clinicopathologic characteristics and disease outcome were performed. Results: BCs with late-stage diagnosis, high histological grade, or high proliferative rate were more likely to be HER2 1+ or 2+/FISH- in comparison with earlier stage, low-grade, or low-proliferative tumors (lower bounds of 95% confidence intervals [CIs] for odds ratios [ORs] > 1). BCs with high expression of ER (≥ 80% [n=12383]) were more enriched with HER2 1+ (OR 1.39; 95%CI, 1.2-1.6) or 2+/FISH- tumors (OR 1.26; 95%CI, 1.0-1.6) than ER-low/moderate (1-79% [n=1144]) ones. The 5-year overall survival (OS) for HER2 1+ BCs was lower than that for HER2 0 or 2+/FISH- tumors (P = 0.0018). This finding was confirmed also after stratification by ER status (low/moderate and high expression; P = 0.07 and 0.007, respectively). By multivariate logistic regression, the following variables were significantly associated with HER2 1+, or 2+/FISH- expression compared to HER2 0 tumors: age at diagnosis < 50, high histological grade, ER expression ≥ 80% (lower bounds of 95% CIs for ORs > 1). Multivariate Cox's regression analysis showed that histological grades G2 and G3 were independently associated with poor survival vs. G1 tumors (HR 1.2; 95%CI, 1.1-1.3 and 1.3; 95%CI, 1.1-1.4, respectively). Furthermore, HER2 2+/FISH- status was significantly associated with better survival in comparison with HER2 0 tumors (HR 0.82; 95%CI, 0.7-0.9). This finding was confirmed also after stratification by ER status (ER 1-79% and ≥ 80%). No significant difference in OS was observed between HER2 1+ and 0 BCs. Conclusions: The putative worse prognostic impact of HER2 expression in pts with ER-positive/HER2-negative BCs was not confirmed. The better outcome observed in pts with HER2 2+/FISH- BCs may be related to the known FISH-negative (HER2-non-amplified) status of this subgroup. These findings may help identify optimal patient inclusion criteria for clinical trials with novel anti-HER2 therapies in ER-positive/HER2-negative disease. Citation Format: Antonino Musolino, Olga Serra, Benedetta Pellegrino, Chiara Tommasi, Daniele Zanoni, Fabio Faccini, Maria Michiara, Paolo Sgargi, Giuliano Carrozzi, Stefano Ferretti, Veruska Grossi, Alessandra Ravaioli, Federica Zamagni, Fabio Falcini. Prognostic Role of HER2 Expression in Patients with ER-positive/HER2-negative Breast Cancer. Results from a Population-Based Cancer Registry Study [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-26-01.

Abstract PO5-07-07: The biological basis of breast MRI background parenchymal enhancement in women with high breast cancer risk

Abstract Introduction: The amount and degree of the fibroglandular tissue (FGT) enhancement on breast MRI, known as background parenchymal enhancement (BPE), is an emerging marker of breast cancer risk. Higher qualitative and quantitative BPE levels have been shown to correlate with both future risk of initial breast cancer diagnosis and breast cancer recurrence after treatment. Furthermore, adjuvant and preventative therapies such as radiation and endocrine therapy have been shown to lower BPE in many women, potentially serving as a marker of treatment efficacy. However, the biological basis of BPE remains unclear and elucidation of this mechanism could identify novel targeted prevention strategies. The goal of this study was to assess the association of BPE levels with markers of inflammation, vascularity, proliferation, and estrogen sensitivity in a cohort of high-risk women. Materials and Methods: In this IRB-approved retrospective study, we identified asymptomatic high-risk women who underwent at least one dynamic contrast-enhanced (DCE) breast MRI within one year of prophylactic bilateral mastectomy. Qualitative BPE assessments were obtained from clinical reports. Quantitative BPE measures were obtained using in-house semi-automated software: whole breast FGT was first segmented on pre-contrast images, percent enhancement (PE) was calculated for each voxel as (S1-S0)/S0 × 100%, where S0 and S1 are pre- and early post-contrast signal intensity, respectively, and quantitative BPE was then defined as the mean PE of the FGT. Additionally, the BPE volume ratio was calculated as the fraction of FGT that had a PE >50%. Histopathology markers of inflammation (COX-2), vascularity (VEGF), proliferation (Ki-67), and estrogen receptor status (ER) were prospectively measured on archived mastectomy specimens of each breast. BPE metrics were correlated with histopathological markers using Spearman’s rank correlation, with and without adjustment by menopausal status of the patient. All tests were based on generalized estimating equations models to account for the non-independence of the two breasts per patient. A p-value < 0.05 was considered significant. Results: We identified 56 women (median age: 39; range: 26-63 years) for this study, of which 39% were post-menopausal (N=22). Distribution of qualitative BPE levels were 41% minimal (N=23), 29% mild (N=16), 18% moderate (N=10), and 12% marked (N=7). Qualitative BPE and quantitative BPE markers were moderately positively correlated (r=0.37-0.54, p< 0.01). Table 1 lists the correlations between BPE metrics and histopathology markers. After adjusting for menopausal status, qualitative BPE was positively associated with ER (r=0.27, p=0.02) and VEGF (r=0.24, p< 0.01; r=0.21, p=0.02 adjusted). Quantitative BPE and BPE volume ratio were both positively correlated with VEGF (p< 0.04) and negatively correlated with COX2 (p< 0.05). No significant associations were observed between BPE and Ki-67. Discussion: Our study suggests that increased BPE on MRI positively correlates with increased vascularity and higher estrogen receptor expression but not inflammation or proliferation within normal breast tissue in high-risk women. These findings, along with established associations between BPE and cancer risk, could be useful to select and assess efficacy of targeted preventative treatments. Table. Spearman rank correlations between BPE metrics and histopathological markers. Citation Format: Pegah Khoshpouri, Anum Kazerouni, Sana Parsian, Daniel Hippe, Olivia Walsh, Lisa Koch, Savannah Partridge, Habib Rahbar. The biological basis of breast MRI background parenchymal enhancement in women with high breast cancer risk [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-07-07.

Abstract PR007: Patient derived organoids as a model to study estrogen mediated endometrial cancer

Abstract The purpose of our study was to develop a more optimal in vitro model to study the mechanisms of hormone action on endometrial cancer. Endometrial cancer (EC) represents a major health disparity with a 21% increase in mortality in African American women. The cause for this excess mortality is unknown. Endometrial cancer is commonly characterized by histological subtype into Endometrioid or Non-Endometrioid with estrogen being a driver for many endometroid type tumors. Most EC cell lines do not have high expression of estrogen receptor or do not respond to physiological levels of hormones in vitro; this lack of functional experimental models has hampered progress in the study of this disease. We have developed Patient Derived Organoids (PDOs) from 9 patient-derived xenografts (PDX) that are from both African American and Non-Hispanic Caucasian women with various stages and subtypes of EC, as a model to study EC. This model system captures the characteristics of different histological and molecular subtypes in vitro, as they maintain not only EC tumor cells but also cells from the adjacent tumor stroma. Our developed PDOs were derived from multiple molecular subtypes of EC that have different characteristics and genomic features, allowing us to observe differences in the basal properties of each tumor in vitro. The use of this organoid model system is advantageous for studying EC because growth characteristics are similar to an in vivo model, with the organoids preserving the genotype and phenotype of the native EC, allowing us to observe cellular interactions. We have observed inter-organoid variation of protein expression of β-catenin, E-cadherin, and vimentin, proteins playing a pivotal role in cellular contact, signaling, and epithelial-mesenchymal transition (EMT). Using qPCR and Western blot analysis we have observed high expression levels of estrogen receptor and progesterone receptor in endometrioid type PDOs to begin the exploration of hormonal signaling in this model. Treatment of these same PDOs with β-estradiol resulted in an increase in proliferation of the stromal cells. Single cell RNA sequencing was performed on two PDO’s that had the highest expression of progesterone and estrogen receptors. This analysis demonstrated cell population heterogeneity with multiple cell clusters forming with differential gene expression. From this analysis we can see that the cells recapitulate the gene expression from the tumor of origin, with cells containing the same genetic mutation as the original patient. These results make this model potentially good for studying hormone signaling cascades seen in EC patients that have not been previously observed in established EC cell lines. These studies will also aid in the opportunity to explore the function of differential cell types in the pathogenesis of this disease. Citation Format: Breanna M.W. Jeffcoat, Andrew B. Galdden, Russell R. Broaddus. Patient derived organoids as a model to study estrogen mediated endometrial cancer [abstract]. In: Proceedings of the AACR Special Conference on Endometrial Cancer: Transforming Care through Science; 2023 Nov 16-18; Boston, Massachusetts. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(5_Suppl):Abstract nr PR007.

Adavosertib-encapsulated metal-organic frameworks for p53-mutated gallbladder cancer treatment via synthetic lethality

Adavosertib (ADA) is a WEE1 inhibitor that exhibits a synthetic lethal effect on p53-mutated gallbladder cancer (GBC). However, drug resistance due to DNA damage response compensation pathways and high toxicity limits further applications. Herein, estrone-targeted ADA-encapsulated metal–organic frameworks (ADA@MOF-EPL) for GBC synthetic lethal treatment by inducing conditional factors are developed. The high expression of estrogen receptors in GBC enables ADA@MOF-EPL to quickly enter and accumulate near the cell nucleus through estrone-mediated endocytosis and release ADA to inhibit WEE1 upon entering the acidic tumor microenvironment. Ultrasound irradiation induces ADA@MOF-EPL to generate reactive oxygen species (ROS), which leads to a further increase in DNA damage, resulting in a higher sensitivity of p53-mutated cancer cells to WEE1 inhibitor and promoting cell death via conditional synthetic lethality. The conditional factor induced by ADA@MOF-EPL further enhances the antitumor efficacy while significantly reducing systemic toxicity. Moreover, ADA@MOF-EPL demonstrates similar antitumor abilities in other p53-mutated solid tumors, revealing its potential as a broad-spectrum antitumor drug.

Combined Effects of Polystyrene Nanosphere and Homosolate Exposures on Estrogenic End Points in MCF-7 Cells and Zebrafish.

Micro- and nanoplastics (MNPs) and homosalate (HMS) are ubiquitous emerging environmental contaminants detected in human samples. Despite the well-established endocrine-disrupting effects (EDEs) of HMS, the interaction between MNPs and HMS and its impact on HMS-induced EDEs remain unclear. This study aimed to investigate the influence of MNPs on HMS-induced estrogenic effects and elucidate the underlying mechanisms in vitro and invivo. We assessed the impact of polystyrene nanospheres (PNSs; , ) on HMS-induced MCF-7 cell proliferation (HMS: , equivalent to ) using the E-SCREEN assay and explored potential mechanisms through transcriptomics. Adult zebrafish were exposed to HMS () with or without PNSs (, ) for 21 d. EDEs were evaluated through gonadal histopathology, fertility tests, steroid hormone synthesis, and gene expression changes in the hypothalamus-pituitary-gonad-liver (HPGL) axis. Coexposure of HMS and PNSs resulted in higher expression of estrogen receptor (ESR1) and the mRNAs of target genes (pS2, AREG, and PGR), a greater estrogen-responsive element transactivation activity, and synergistic stimulation on MCF-7 cell proliferation. Knockdown of serum and glucocorticoid-regulated kinase 1 (SGK1) rescued the MCF-7 cell proliferation induced by PNSs alone or in combination with HMS. In zebrafish, coexposure showed higher expression of SGK1 and promoted ovary development but inhibited spermatogenesis. In addition, coexposure led to lower egg hatchability, higher embryonic mortality, and greater larval malformation. Coexposure also modulated steroid hormone synthesis genes (cyp17a2, hsd17[Formula: see text]1, esr2b, vtg1, and vtg2), and resulted in higher () release in females. Conversely, males showed lower testosterone, , and gene expressions of cyp11a1, cyp11a2, cyp17a1, cyp17a2, and hsd17[Formula: see text]1. PNS exposure exacerbated HMS-induced estrogenic effects via SGK1 up-regulation in MCF-7 cells and disrupting the HPGL axis in zebrafish, with gender-specific patterns. This offers new mechanistic insights and health implications of MNP and contaminant coexposure. https://doi.org/10.1289/EHP13696.

Optimization of small molecule degraders and antagonists for targeting estrogen receptor based on breast cancer: current status and future.

The estrogen receptor (ER) is a classical receptor protein that plays a crucial role in mediating multiple signaling pathways in various target organs. It has been shown that ER-targeting therapies inhibit breast cancer cell proliferation, enhance neuronal protection, and promote osteoclast formation. Several drugs have been designed to specifically target ER in ER-positive (ER+) breast cancer, including selective estrogen receptor modulators (SERM) such as Tamoxifen. However, the emergence of drug resistance in ER+ breast cancer and the potential side effects on the endometrium which has high ER expression has posed significant challenges in clinical practice. Recently, novel ER-targeted drugs, namely, selective estrogen receptor degrader (SERD) and selective estrogen receptor covalent antagonist (SERCA) have shown promise in addressing these concerns. This paper provides a comprehensive review of the structural functions of ER and highlights recent advancements in SERD and SERCA-related small molecule drugs, especially focusing on their structural optimization strategies and future optimization directions. Additionally, the therapeutic potential and challenges of novel SERDs and SERCAs in breast cancer and other ER-related diseases have been discussed.

Prevalence of ESR1 gene mutations in gynecologic malignancies with high-estrogen receptor expression (1283)

Prevalence of ESR1 gene mutations in gynecologic malignancies with high-estrogen receptor expression (1283)

Abstract P4-02-14: Gain of HER2 Amplification in Patients with HR+/HER2- and Triple Negative Early Breast Cancer Treated with Neoadjuvant Chemotherapy

Abstract Background: Neoadjuvant chemotherapy (NAC) is standard of care for the majority of patients with clinical stage II-III triple negative breast cancer (TNBC) and is considered in high-risk patients with hormone receptor positive (HR+)/human epidermal growth factor receptor 2 (HER2) negative (-) tumors, with expected pathological complete response (pCR) rates of 40-60% and 10-12%, respectively. In HER2- patients with residual disease (RD) after NAC, there is limited data on rates of gain of HER2 amplification. The biological and clinical significance of this phenomenon is unknown and determining the best adjuvant therapy for these patients remains a challenge. We sought to determine the rate of HER2 gain in a cohort of consecutive patients with HER2- breast cancer (BC) treated with NAC. Methods: From 01/2021 to 12/2021, we identified patients with HER2- breast cancer treated with NAC followed by surgery at our institution. Patients who received neoadjuvant endocrine therapy were excluded. The rates of pCR (ypT0/is ypN0) and HER2 status pre- and post-NAC were assessed. Estrogen receptor (ER), progesterone receptor (PR) and HER2 status on surgery specimens were internally determined for all patients using ASCO/CAP 2020 guidelines. ER-low was defined as ER expression by immunohistochemistry (IHC) 1-10%. Results: We included 256 patients, 130 (51%) HR+/HER2- [13/130(10%) ER-low] and 126 (49%) TNBC. Median age was 48 years (range 25-82) and the majority presented with clinical T2 (57%) and N1 (59%). Of 130 patients with HR+/HER2-tumors, 120 (92%) received dose-dense (dd) doxorubicin/cyclophosphamide-paclitaxel (AC-T). Of 126 TNBC patients, 46 (37%) received ddAC followed by carboplatin in combination with paclitaxel +/- pembrolizumab. Centralized HER2 status assessment on the core biopsy was performed in 22% of samples. Overall, pCR was achieved in 40% of TNBC and 11% of HR+/HER2-. Among the 192 patients with RD, the rate of HER2 gain was 8/192 (4%), including 3% (2/76) of TNBC and 5% (6/116) of HR+/HER2- patients. 7 of the 8 patients (88%) converted from IHC 1+ or 2+ fluorescence in situ hybridization (FISH) not amplified on core biopsy to IHC 2+ FISH amplified on the surgical specimen. In only 1 case, the HER2 status converted from IHC2+ FISH not amplified to IHC3+. 3/8 patients had multifocal disease. All 6 patients with HR+/HER2- BC and HER2 gain had high (>90%) ER expression (Table 1). All but one patient with HER2 gain received adjuvant anti-HER2 therapy. After a median follow-up of 10 months, no recurrence events occurred in this group.12 of the remaining 184 patients experienced a recurrence [11 distant recurrences (8 and 3, in the TNBC and HR+/HER2- groups, respectively), and there was 1 local event (localized chest wall recurrence) in the HR+/HER2- group]. Conclusions: At a single center, we found that in patients with HER2- BC treated with NAC, HER2 gain in patients with RD was uncommon and occurred more frequently in those with HR+ tumors. Analysis of a larger cohort is ongoing to corroborate these results. It is remains to be determined if this phenomenon represents a true HER2 status conversion or tumor heterogeneity. Table 1: Clinico-pathological characteristics of patients with HER2 gain on residual disease Citation Format: Emanuela Ferraro, Sonya Chew Minmin, Anton Safonov, Andrea V. Barrio, Shanu Modi, Andrew D Seidman, Hanna Y Wen, Edi Brogi, Mark E. Robson, Chau T Dang. Gain of HER2 Amplification in Patients with HR+/HER2- and Triple Negative Early Breast Cancer Treated with Neoadjuvant Chemotherapy [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P4-02-14.

Clinicopathologic Characteristics and Prognosis of ERBB2-Low Breast Cancer Among Patients in the National Cancer Database

Given conflicting results regarding the prognosis of erb-b2 receptor tyrosine kinase 2 (ERBB2; formerly HER2 or HER2/neu)-low breast cancer, a large-scale, nationally applicable comparison of ERBB2-low vs ERBB2-negative breast cancer is needed. To investigate whether ERBB2-low breast cancer is a clinically distinct subtype in terms of epidemiological characteristics, prognosis, and response to neoadjuvant chemotherapy. This retrospective cohort study was conducted using the National Cancer Database, including 1 136 016 patients in the US diagnosed with invasive breast cancer from January 1, 2010, to December 31, 2019, who had ERBB2-negative disease and had immunohistochemistry results available. ERBB2-low tumors were classified as having an immunohistochemistry score of 1+, or 2+ with a negative in situ hybridization test. Data were analyzed from November 1, 2021, through November 30, 2022. Standard therapy according to routine clinical practice. The primary outcomes were overall survival (OS), reported as adjusted hazard ratios (aHRs), and pathologic complete response, reported as adjusted odds ratios (aORs), for ERBB2-negative vs ERBB2-low breast cancer, controlling for age, sex, race and ethnicity, Charlson-Deyo Comorbidity Index score, treatment facility type, tumor grade, tumor histology, hormone receptor status, and cancer stage. The study identified 1 136 016 patients (mean [SD] age, 62.4 [13.1] years; 99.1% female; 78.6% non-Hispanic White), of whom 392 246 (34.5%) were diagnosed with ERBB2-negative and 743 770 (65.5%) with ERBB2-low breast cancer. The mean (SD) age of the ERBB2-negative group was 62.1 (13.2) years and 62.5 (13.0) years for the ERBB2-low group. Higher estrogen receptor expression was associated with increased rates of ERBB2-low disease (aOR, 1.15 per 10% increase). Compared with non-Hispanic White patients, of whom 66.1% were diagnosed with ERBB2-low breast cancer, fewer non-Hispanic Black (62.8%) and Hispanic (61.0%) patients had ERBB2-low disease, although in non-Hispanic Black patients this was mediated by differences in rates of triple-negative disease and other confounders. A slightly lower rate of pathologic complete response was seen in patients with ERBB2-low disease vs patients with ERBB2-negative disease on multivariable analysis (aOR, 0.89; 95% CI, 0.86-0.92; P < .001). ERBB2-low status was also associated with small improvements in OS for stage III (aHR, 0.92; 95% CI, 0.89-0.96; P < .001) and stage IV (aHR, 0.91; 95% CI, 0.87-0.96; P < .001) triple-negative breast cancer, although this amounted to only a 2.0% (stage III) and 0.4% (stage IV) increase in 5-year OS. This large-scale retrospective cohort analysis found minimal prognostic differences between ERBB2-low and ERBB2-negative breast cancer. These findings suggest that, moving forward, outcomes in ERBB2-low breast cancer will be driven by ERBB2-directed antibody-drug conjugates, rather than intrinsic differences in biological characteristics associated with low-level ERBB2 expression. These findings do not support the classification of ERBB2-low breast cancer as a unique disease entity.

E3 Ubiquitin Ligase NEDD4 Affects Estrogen Receptor α Expression and the Prognosis of Patients with Hormone Receptor-Positive Breast Cancer.

Neural precursor cell-expressed developmentally downregulated 4-1 (NEDD4) is an E3 ligase that leads to the degradation of proteins, including estrogen receptor α. We evaluated whether the expression level of NEDD4 affected the outcome of breast cancer patients. We performed a retrospective cohort study enrolling 143 patients with hormone receptor-positive, human epidermal growth factor receptor 2-negative early breast cancer. Of the 66 patients with high NEDD4 mRNA levels (high NEDD4 group) and 77 patients with low NEDD4 mRNA levels (low NEDD4 group), 98.4% and 96.1%, respectively, of the patients had received neoadjuvant/adjuvant hormone therapy. Disease-free survival and overall survival were significantly longer in the low NEDD4 group than in the high NEDD4 group (p = 0.048 and p = 0.022, respectively). Western blotting revealed a high expression of estrogen receptor α in the NEDD4-knockdown culture cells. The proliferation of NEDD4-knockdown cells treated with tamoxifen or estradiol deprivation was suppressed, compared with that of NEDD4-expressing cells. Knockdown of NEDD4 in breast cancer cells induced the accumulation of estrogen receptor α and increased sensitivity to hormone therapy. In summary, this mechanism may lead to a better prognosis in hormone receptor-positive breast cancer patients with a low expression of NEDD4.

Transcriptional Profile Associated with Clinical Outcomes in Metastatic Hormone-Sensitive Prostate Cancer Treated with Androgen Deprivation and Docetaxel.

Simple SummaryThe combination of androgen deprivation therapy (ADT) with docetaxel (DX) or/and with novel anti-androgen receptor therapies have become standards for the treatment of patients with metastatic hormone-sensitive prostate cancer (mHSPC). However, metastatic PC remains incurable, and biomarkers for individual treatment selection are needed. We propose here that molecular alterations associated with castration resistance may predict the clinical evolution of mHSPC patients. To test this hypothesis, we designed a custom expression panel of 184 genes and tested it in tumor biopsies from patients with mHSPC treated with ADT+DX. We found that AR and ESR signatures and ESR2 gene expression correlate with a good prognosis. The lower expression of TSG (PTEN, TP53 and RB1) signature, as well as high ARV7 and low RB1 gene expression, were associated with adverse clinical outcomes. The usefulness of transcriptomic analysis of such signatures as a strategy for personalized treatment selection should be further explored.(1) Background: Androgen deprivation therapy (ADT) and docetaxel (DX) combination is a standard therapy for metastatic hormone-sensitive prostate cancer (mHSPC) patients. (2) Methods: We investigate if tumor transcriptomic analysis predicts mHSPC evolution in a multicenter retrospective biomarker study. A customized panel of 184 genes was tested in mRNA from tumor samples by the nCounter platform in 125 mHSPC patients treated with ADT+DX. Gene expression was correlated with castration-resistant prostate cancer-free survival (CRPC-FS) and overall survival (OS). (3) Results: High expression of androgen receptor (AR) signature was independently associated with longer CRPC-FS (hazard ratio (HR) 0.6, 95% confidence interval (CI) 0.3–0.9; p = 0.015), high expression of estrogen receptor (ESR) signature with longer CRPC-FS (HR 0.6, 95% CI 0.4–0.9; p = 0.019) and OS (HR 0.5, 95% CI 0.2–0.9, p = 0.024), and lower expression of tumor suppressor genes (TSG) (RB1, PTEN and TP53) with shorter OS (HR 2, 95% CI 1–3.8; p = 0.044). ARV7 expression was independently associated with shorter CRPC-FS (HR 1.5, 95% CI 1.1–2.1, p = 0.008) and OS (HR 1.8, 95% CI 1.2–2.6, p = 0.004), high ESR2 was associated with longer OS (HR 0.5, 95% CI 0.2–1, p = 0.048) and low expression of RB1 was independently associated with shorter OS (HR 1.9, 95% CI 1.1–3.2, p = 0.014). (4) Conclusions: AR, ESR, and TSG expression signatures, as well as ARV7, RB1, and ESR2 expression, have a prognostic value in mHSPC patients treated with ADT+DX.

Spatial oriented single-cell proteomic assay of low grade serous ovarian carcinoma identifies tumor oncogenic pathways associated with brain metastases (233)

Objectives: As the systemic treatment of epithelial ovarian cancer (EOC) has improved, and the survival is longer, the incidence of brain metastases (BM) in EOC patients has increased to about 2-6% and invariably results in death. In order to better understand the molecular mechanisms involved in the development of EOC BM and to characterize the BM architecture and heterogeneity, we recently used multiplex cyclic-immunofluorescence (Cyc-IF) to characterize a set of matched samples from a single ovarian cancer patient. Methods: The samples included a primary tumor, a peritoneal tumor collected at recurrence, and a BM. Cyc-IF is a spatial oriented single-cell proteomic assay that allows the quantification of over 60 proteins in a single FFPE slide. Results: Our analyses indicated that tumor architecture and heterogeneity were highly altered in the BM compared to the primary and recurrent tumors in this patient. Indeed, the primary and recurrent intraperitoneal tumors showed extensive heterogeneity with cancer cells in different cell states related to the cell cycle (cyclins), hormone receptors (PR and ER) and signaling pathways. In contrast, the BM was overall more homogeneous, with no PR receptor expression and low expression of cyclins. EOC cells from the BM harbor higher oncogenic MAPK and PI3K-AKT signaling pathway activity, higher estrogen receptor expression and displayed immunosuppressive properties including increased expression of PD1 and the presence of tumor-associated macrophages (TAM) and regulatory T cells (Tregs). Importantly, we were able to identify cell populations in the primary tumor that had phenotypes similar to that of cells found in the BM. Conclusions: These findings are consistent with the concept that EOC cells with a predilection for BM are present early during the disease course. Therefore, this represents an opportunity to develop a molecular signature for BM prevention and treatment. Objectives: As the systemic treatment of epithelial ovarian cancer (EOC) has improved, and the survival is longer, the incidence of brain metastases (BM) in EOC patients has increased to about 2-6% and invariably results in death. In order to better understand the molecular mechanisms involved in the development of EOC BM and to characterize the BM architecture and heterogeneity, we recently used multiplex cyclic-immunofluorescence (Cyc-IF) to characterize a set of matched samples from a single ovarian cancer patient. Methods: The samples included a primary tumor, a peritoneal tumor collected at recurrence, and a BM. Cyc-IF is a spatial oriented single-cell proteomic assay that allows the quantification of over 60 proteins in a single FFPE slide. Results: Our analyses indicated that tumor architecture and heterogeneity were highly altered in the BM compared to the primary and recurrent tumors in this patient. Indeed, the primary and recurrent intraperitoneal tumors showed extensive heterogeneity with cancer cells in different cell states related to the cell cycle (cyclins), hormone receptors (PR and ER) and signaling pathways. In contrast, the BM was overall more homogeneous, with no PR receptor expression and low expression of cyclins. EOC cells from the BM harbor higher oncogenic MAPK and PI3K-AKT signaling pathway activity, higher estrogen receptor expression and displayed immunosuppressive properties including increased expression of PD1 and the presence of tumor-associated macrophages (TAM) and regulatory T cells (Tregs). Importantly, we were able to identify cell populations in the primary tumor that had phenotypes similar to that of cells found in the BM. Conclusions: These findings are consistent with the concept that EOC cells with a predilection for BM are present early during the disease course. Therefore, this represents an opportunity to develop a molecular signature for BM prevention and treatment.

Correlations between dynamic-enhanced magnetic resonance imaging quantitative parameters and postoperative recurrence or metastasis and clinicopathological features in breast cancer patients-a retrospective cohort study.

BackgroundFew related studies focused on the correlations between the quantitative parameters of dynamic-enhanced magnetic resonance imaging (MRI) and the clinical pathological characteristics of patients with invasive breast cancer have been conducted to date. This study sought to explore the value of quantitative parameters of dynamic-enhanced MRI in predicting postoperative recurrence or metastasis in breast cancer patients and their correlations with clinical pathological features, so as to provide clinicians with understanding of MRI in breast cancer.MethodsFrom January 2016 to June 2017, 214 invasive breast cancer patients admitted to Affiliated Kunshan Hospital of Jiangsu University were retrospectively enrolled in this study. Dynamic-enhanced MRI was performed to analyze the relationship between quantitative parameters of dynamic-enhanced MRI and recurrence or metastasis, and analyze their correlations with clinical pathological features in patients with invasive breast cancer.ResultsThe apparent diffusion coefficient and peak time had certain diagnostic value for postoperative recurrence or metastasis in breast cancer patients, and the areas under the curve were 0.821 [95% confidence interval (CI): 0.732–0.911; P<0.001] and 0.691 (95% CI: 0.609–0.774; P<0.001), respectively. An apparent diffusion coefficient <0.78×10−3 mm2/s, a peak time <167.50 s, tumor staging (T staging) ≥2, vascular tumor thrombus, and positive lymph nodes were risk factors for postoperative recurrence or metastasis in breast cancer patients (odds ratio: 19.768, 95% CI: 2.577–151.619, P=0.004; 5.708, 95% CI: 1.088–29.947, P=0.039; 122.474, 95% CI: 5.334–2,812.360, P=0.003; 28.304, 95% CI: 1.372–583.914, P=0.030; 314.407, 95% CI: 10.617–9,310.547, P=0.001), and high estrogen receptor (ER) expression was a protective factor for postoperative recurrence or metastasis in breast cancer patients (odds ratio: 0.056, 95% CI: 0.004–0.795, P=0.033). The apparent diffusion coefficient was related to the site of onset, T staging, vascular tumor thrombus, and positive lymph nodes in breast cancer patients (P<0.05). Peak time was related to a high nuclear-associated antigen Ki-67 index, high ER expression, and high progesterone receptor (PR) expression in breast cancer patients (P<0.05).ConclusionsThe quantitative parameters of MRI were associated with clinical pathological characteristics and recurrence or metastasis in breast cancer after surgery.

Promising Therapeutic Impact of a Selective Estrogen Receptor Downregulator, Fulvestrant, as Demonstrated In Vitro upon Low-Grade Serous Ovarian Carcinoma Cell Lines.

Few studies have reported hormonal agent use in the treatment of low-grade serous ovarian carcinomas (LGSOCs), which are chemoresistant. Considering the need for novel effective therapies, we investigated the hormone receptor expression and hormonal inhibition efficacy in LGSOCs. Using immunohistochemistry, we assessed the estrogen receptor (ER) expression status in 33 cases of histologically confirmed serous ovarian tumors, including 10, 11, and 12 cases of LGSOCs, serous borderline tumors (SBTs), and serous cystadenomas (SCAs), respectively. The genetic background reported in our previous study was used in the current study. MPSC1 cells, which were established from LGSOCs, were used in cell proliferation assays. We observed a higher ER expression in LGSOCs and SBTs than in SCAs (70%, 81%, and 50%, respectively). Thus, LGSOCs and SBTs exhibit higher ER expression than SCAs. Moreover, the PIK3CA mutation positively correlated with ER expression in LGSOCs (p = 0.0113). MPSC1 cells showed low ER expression on Western blotting. MPSC1 cell proliferation was significantly inhibited by fulvestrant (a selective ER downregulator). The activation of ER and PI3K/AKT signaling pathways may play an important role in LGSOC carcinogenesis. ER downregulation with fulvestrant or combination therapy with PI3K inhibitors is a possible novel treatment for patients with LGSOCs.

Long non-coding RNA LINC02613 is a prognostic biomarker for breast cancer and correlates with the cell cycle and immune infiltration based on TCGA data.

BackgroundThe development of transcriptomics has provided a new perspective to understand cancers. However, the role of long non-coding RNAs (lncRNAs) has not been fully elucidated. In the field of breast cancer, exploring the biological role and impact of lncRNAs on patient prognosis is of great importance.MethodsExpression data of the lncRNA, LINC02613, were downloaded from The Cancer Genome Atlas (TCGA) dataset. The Kruskal-Wallis rank sum test, Wilcoxon rank sum test and logistic regression analysis were used to investigate the relationship between LINC02613 and clinicopathological characteristics. Cox regression and Kaplan-Meier analyses were used to assess the prognostic value of LINC02613. Gene ontology (GO) analysis, gene set enrichment analysis (GSEA) and immune infiltration analysis were used to explore the potential biological mechanisms of LINC02613.ResultsThe expression of LINC02613 was downregulated in tumor samples and was related to poor overall survival (OS) in breast cancer patients. Statistical analysis revealed that low expression of LINC02613 was associated with Asian ethnicity, older age, higher histological grade and higher expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). Positive expression of immunohistochemical markers (ER, PR and HER2), high pathological stage and N stage were independent predictors for low LINC02613 expression. Decreased expression of LINC02613 caused an increase in Wnt2 and Wnt3 protein expression, which activated the Wnt signaling pathway, thereby promoting cell cycle progression, DNA replication and glycolysis. At the same time, the antitumor immunity of the body was also weakened when the expression of LINC02613 was decreased.ConclusionsLow expression of LINC02613 predicts poor OS in breast cancer patients. LINC02613 is involved in the regulation of the cell cycle, DNA replication and glycolysis via the Wnt signaling pathway, and it is related to antitumor immunity.

EZH2 Mediates Proliferation, Migration, and Invasion Promoted by Estradiol in Human Glioblastoma Cells.

Glioblastomas (GBM) are the most frequent and aggressive brain tumors. 17β-estradiol (E2) increases proliferation, migration, and invasion of human GBM cells; however underlying mechanisms are no fully understood. Zeste 2 Enhancer Homologous enzyme (EZH2) is a methyltransferase part of Polycomb 2 repressor complex (PRC2). In GBM, EZH2 is overexpressed and involved in the cell cycle, migration, and invasion processes. We studied the role of EZH2 in the pro-oncogenic actions of E2 in human GBM cells. EZH2 gene silencing and pharmacological inhibition of EZH2 blocked proliferation, migration, and invasion of GBM cells induced by E2. We identified in silico additional putative estrogen response elements (EREs) at the EZH2 promoter, but E2 did not modify EZH2 expression. In silico analysis also revealed that among human GBM samples, EZH2 expression was homogeneous; in contrast, the heterogeneous expression of estrogen receptors (ERs) allowed the classification of the samples into groups. Even in the GBM cluster with high expression of ERs and those of their target genes, the expression of PCR2 target genes did not change. Overall, our data suggest that in GBM cells, pro-oncogenic actions of E2 are mediated by EZH2, without changes in EZH2 expression and by mechanisms that appear to be unrelated to the transcriptional activity of ERs.

Bortezomib-Encapsulated Dual Responsive Copolymeric Nanoparticles for Gallbladder Cancer Targeted Therapy.

Gallbladder cancer (GBC) is a rare but the most malignant type of biliary tract tumor. It is usually diagnosed at an advanced stage and conventional treatments are unsatisfactory. As a proteasome inhibitor, bortezomib (BTZ) exhibits excellent antitumor ability in GBC. However, the long‐term treatment efficacy is limited by its resistance, poor stability, and high toxicity. Herein, BTZ‐encapsulated pH‐responsive copolymeric nanoparticles with estrone (ES‐NP(BTZ; Ce6)) for GBC‐specific targeted therapy is reported. Due to the high estrogen receptor expression in GBC, ES‐NP(BTZ; Ce6) can rapidly enter the cells and accumulate near the nucleus via ES‐mediated endocytosis. Under acidic tumor microenvironment (TME) and 808 nm laser irradiation, BTZ is released and ROS is generated by Ce6 to destroy the “bounce‐back” response pathway proteins, such as DDI2 and p97, which can effectively inhibit proteasomes and increase apoptosis. Compared to the traditional treatment using BTZ monotherapy, ES‐NP(BTZ; Ce6) can significantly impede disease progression at lower BTZ concentrations and improve its resistance. Moreover, ES‐NP(BTZ; Ce6) demonstrates similar antitumor abilities in patient‐derived xenograft animal models and five other types of solid tumor cells, revealing its potential as a broad‐spectrum antitumor formulation.

A pilot analysis of headache disorders in breast cancer patients.

The interaction between breast cancer and migraine is complex and not fully elucidated. Large epidemiological studies point towards a beneficial effect of migraines on breast cancer (BC). We aimed to investigate the BC-migraine relationship, with strict data checks and clinical evaluations of both BC and common headache forms. Consecutive BC patients were evaluated with the International Classification of Headache Disorders. Clinical data on the BC subtypes and treatments were collected. Parametric and nonparametric statistics were used according to data distributions. Fifty patients were recruited. The mean age was 53.5 ± 12.5years; 42% were postmenopausal, 52% were premenopausal, and 6% were peri-menopausal. Eleven patients were diagnosed as luminal A, nine as luminal B, 24 as HER2-positive (HER2 +), six as triple-negative BC. Thirty-eight (76%) patients had hormone receptor-positive disease. Ninety-twopercent received chemotherapy, 66% received endocrine therapy, and 52% received radiotherapy. Nine out of 50 reported a worsening of headache after systemic treatment. Migraine was diagnosed in 29 patients (18 with menstrual migraine), tension-type headache (TTH) in nine, and no headache in 12. Patients with migraine were younger (48.4 ± 10.7 vs. 60.5 ± 12; p < 0.01). Patients with migraine and TTH had a higher chance of having a HER2 + BC (p < 0.05). Active migraine was associated with a higher expression of estrogen receptors (p = 0.04). Patients with active migraine had higher estrogen receptor expression, while migraine and TTH patients mainly had HER2 + BC. This association was not known earlier and could be helpful to understand deeper the relationship between BC and headache.

Neoadjuvant Endocrine Therapy for Operable Breast Cancer: A Retrospective Analysis of Real-World Use.

A retrospective study of the real-world use of neoadjuvant endocrine therapy (NET) is important for standardizing the role of NET in breast cancer care. In a consecutive series of women with operable breast cancer who received NET for ≥28 days, associations of NET objectives, NET outcomes, adjuvant chemotherapy use after NET, and survival with clinicopathological factors were examined. NET objectives were reduction in surgical extent in 49 patients, avoidance of surgery in 31, and treatment until scheduled surgery in 8. The mean duration of NET was 349.5 (range, 34-1,923), 869.8 (range, 36-4,859), and 55.8 (range, 39-113) days, respectively, in these cohorts (success rate: 79.6%, 64.5%, and 100%, respectively), and the differences were significant. Among patients in the former two cohorts, progression-free survival was significantly better in patients with stage 0 or I disease, ductal carcinoma in situ or invasive ductal carcinoma, ≥71% estrogen receptor (ER) positivity, and the surgical extent reduction cohort than the other counterparts. Postoperative chemotherapy use was significantly associated with lymph node metastasis, a high Ki67 labeling index, lymphovascular invasion, and a high preoperative endocrine prognostic index at the time of surgery after NET. Better recurrence-free survival after surgery was significantly associated with high ER expression after NET or high progesterone receptor expression before or after NET. NET can help reduce surgical extent or avoid surgery in women with early breast cancer, ductal carcinoma, or high ER expression. NET may also aid in decisions related to postoperative systemic therapy to improve survival.

Optimal Treatment Duration of Neoadjuvant Endocrine Therapy for Women Aged 60 Years or Older with Estrogen Receptor-Positive, HER2-Negative Invasive Breast Cancer.

Neoadjuvant endocrine therapy is not the standard of care for breast cancer, primarily because the optimal treatment duration remains unclear. This phase 2 prospective multicenter study analyzed time to progression, time to maximal response, and time to treatment failure for neoadjuvant exemestane. Inclusion criteria were women aged ≥60 years with Stage II or III breast cancer classified as estrogen receptor-positive/human epidermal growth factor receptor 2-negative. Response was defined as a ≥10% and minimum of 3 mm decrease in tumor size, as compared with the most recent or smallest value, and no new lesion. Progression was defined as a >10% and minimum of over 3 mm increase in tumor size, as compared with the most recent or smallest value, or a new lesion. Maximal response was defined as the final recorded response. This study included 24 women, most of whom had T2 N0 tumors with high estrogen receptor expression. We initially observed a response in 23 patients (96%); however, 6 patients (25%) later experienced progression. Time to progression, time to maximal response, and time to treatment failure ranged from 7 to 22 months (estimated median, 35), 1 to 22 months (estimated median, 10), and 2 to 22 months (estimated median, 22), respectively. Treatment duration varied widely, but the estimated optimal duration of neoadjuvant exemestane therapy was 22 to 35 months in patients seeking to avoid surgery and 10 months in patients wishing to receive breast-conserving surgery. Neoadjuvant exemestane therapy is long effective for older women with hormone-sensitive breast cancer.