ER-negative Group Research Articles

Immune and Gene-expression Profiling in Estrogen Receptor Low and Negative Early Breast Cancer.

The cut-off of < 1% positive cells to define estrogen receptor (ER) negativity by immunohistochemistry (IHC) in breast cancer (BC) is debated. We explored the tumor immune microenvironment and gene-expression profile of patients with early-stage HER2-negative ER-low (ER 1-9%) BC, comparing them to ER-negative (ER < 1%) and ER-intermediate (ER 10-50%) tumors. Among 921 patients with early-stage I-III, ER ≤ 50%, HER2-negative BCs, tumors were classified as ER-negative (n = 712), ER-low (n = 128), or ER-intermediate (n = 81). Tumor-infiltrating lymphocytes (TILs) were evaluated. CD8+, FOXP3+ cells, and PD-L1 status were assessed by IHC and quantified by digital pathology. We analyzed 776 BC-related genes in 116 samples. All tests were 2-sided at < 0.05 significance level. ER-low and ER-negative tumors exhibited similar median TILs, significantly higher than ER-intermediate tumors. CD8/FOXP3 ratio and PD-L1 positivity rates were comparable between ER-low and ER-negative groups. These groups showed similar enrichment in Basal-like intrinsic subtypes and comparable expression of immune-related genes. ER-low and ER-intermediate tumors showed significant transcriptomic differences. High TILs (≥30%) were associated with improved relapse-free survival (RFS) in ER-low (5-year RFS 78.6% vs 66.2%, log-rank p = .033, hazard ratio (HR) 0.37 [95% CI 0.15-0.96]) and ER-negative patients (5-year RFS 85.2% vs 69.8%, log-rank p < .001, HR 0.41 [95% CI 0.27-0.60]). ER-low and ER-negative tumors are similar biological and molecular entities, supporting their comparable clinical outcomes and treatment responses, including to immunotherapy. Our findings contribute to the growing evidence calling for a reevaluation of ER-positive BC classification and management, aligning ER-low and ER-negative tumors more closely.

Insights into the performance of PREDICT tool in a large Mainland Chinese breast cancer cohort: a comparative analysis of versions 3.0 and 2.2.

PREDICT is a web-based tool for forecasting breast cancer outcomes. PREDICT version 3.0 was recently released. This study aimed to validate this tool for a large population in mainland China and compare v3.0 with v2.2. Women who underwent surgery for nonmetastatic primary invasive breast cancer between 2010 and 2020 from the First Affiliated Hospital of Wenzhou Medical University were selected. Predicted and observed 5-year overall survival (OS) for both v3.0 and v2.2 were compared. Discrimination was compared using receiver-operator curves and DeLong test. Calibration was evaluated using calibration plots and chi-squared test. A difference greater than 5% was deemed clinically relevant. A total of 5424 patients were included, with median follow-up time of 58 months (IQR 38-89 months). Compared to v2.2, v3.0 did not show improved discriminatory accuracy for 5-year OS (AUC: 0.756 vs 0.771), same as ER-positive and ER-negative patients. However, calibration was significantly improved in v3.0, with predicted 5-year OS deviated from observed by -2.0% for the entire cohort, -2.9% for ER-positive and -0.0% for ER-negative patients, compared to -7.3%, -4.7% and -13.7% in v2.2. In v3.0, 5-year OS was underestimated by 9.0% for patients older than 75 years, and 5.8% for patients with micrometastases. Patients with distant metastases postdiagnosis was overestimated by 10.6%. PREDICT v3.0 reliably predicts 5-year OS for the majority of Chinese patients with breast cancer. PREDICT v3.0 significantly improved the predictive accuracy for ER-negative groups. Furthermore, caution is advised when interpreting 5-year OS for patients aged over 70, those with micrometastases or metastases postdiagnosis.

Efficacy of neoadjuvant therapy on HER2-positive breast cancer: a clinicopathological analysis

Objective: To investigate the efficacy of neoadjuvant therapy (NAT) on HER2-positive breast cancer and to analyze their clinicopathological features. Methods: A total of 480 cases of HER2-positive breast cancer who received neoadjuvant therapy (NAT), diagnosed at the Department of Pathology of Fudan University Shanghai Cancer Center from 2015 to 2020, were retrospectively identified. Clinicopathological parameters such as age, tumor size, molecular subtype, type of targeted therapy, Ki-67 proliferation index, ER and HER2 immunohistochemical expression, and HER2 amplification status were analyzed to correlate with the efficacy of NAT. Results: Among 480 patients with HER2-positive breast cancer, 209 achieved pathology complete response (pCR) after NAT, with a pCR rate of 43.5%. Of all the cases,457 patients received chemotherapy plus trastuzumab and 23 patients received chemotherapy with trastuzumab and pertuzumab. A total of 198 cases (43.3%) achieved pCR in patients with chemotherapy plus trastuzumab, and 11 cases (47.8%) achieved pCR in patients with chemotherapy plus trastuzumab and pertuzumab. The pCR rate in the latter group was higher, but there was no statistical significance. The results showed that the pCR rate of IHC-HER2 3+patients (49%) was significantly higher than that of IHC-HER2 2+patients (26.1%, P<0.001). The higher the mean HER2 copy number in the FISH assay, the higher the pCR rate was achieved. The expression level of ER was inversely correlated with the efficacy of NAT, and the pCR rate in the ER-positive group (28.2%) was significantly lower than that in the ER-negative group (55.8%, P<0.001). The pCR rate (29.1%) of patients with luminal B type was lower than that of HER2 overexpression type (55.8%, P<0.001). In addition, higher Ki-67 proliferation index was associated with higher pCR rate (P<0.001). The pCR rate was the highest in the tumor ≤2 cm group (57.7%), while the pCR rate in the tumor >5 cm group was the lowest (31.1%). The difference between the groups was significant (P=0.005). Conclusions: HER2 copy numbers, HER2 immunohistochemical expression level, molecular subtype, ER expression level and Ki-67 proliferation index are significantly associated with pCR after NAT. In addition, fluorescence in situ hybridization results, HER2/CEP17 ratio and tumor size could also significantly affect the efficacy of NAT.

Impact of estrogen receptor expression level on response to neoadjuvant chemotherapy and prognosis in HER2-negative breast cancers

BackgroundBreast cancers with 1–10% cell staining for estrogen receptor (ER) present particular clinical features. The clinical data of estrogen receptor expression level and treatment effect are limited, particularly regarding chemotherapy benefit. We evaluated the pathologic response to neoadjuvant chemotherapy (NAC) in ER low positive tumors (ER staining 1-10%) and compared it with ER > 10% positive tumors (ER staining > 10%) and ER-negative tumors. We further explored the differences in recurrence and survival with respect to the ER expression level.MethodPatients with stages II and III HER2-negative primary breast cancer who received neoadjuvant chemotherapy followed by definitive surgery were categorized according to their ER percentages into three groups: ER-negative, ER low positive, and ER > 10% positive. Logistic regression models were used to assess the association between each variable and pathologic complete response (pCR). Kaplan‒Meier analysis was used to estimate survival outcomes. Cox models were used to adjust for patient and tumor characteristics.ResultsA total of 241 patients were analyzed. Of all patients included, 22 (9.1%) had ER low positive tumors, 159 (66.0%) had ER > 10% positive tumors, and 60 (24.9%) were ER-negative. Low ER positivity was significantly associated with a higher pCR rate than ER > 10% positivity (OR, 0.249; 95% CI, 0.067–0.923; P = 0.038). After a median follow-up time of 32 months, the disease-free survival (DFS) and overall survival (OS) of the patients with ER low positive tumors were significantly worse than those of the patients with ER > 10% positive tumors but similar to those with ER-negative tumors. After adjustment for covariates, ER low positive tumors were significantly associated with worse DFS than ER > 10% positive tumors.ConclusionOur results indicated that ER low positive breast cancer presents a better response to neoadjuvant chemotherapy and significantly worse prognosis for patients than those with ER > 10% positive tumors, but similar to the ER-negative group. These data support that this category of patients behaves clinically like patients with ER-negative breast cancer and should be treated differently from patients with ER > 10% positive tumors. Further prospective study is needed.

Prognostic significance of estrogen receptor status in de novo metastatic breast cancer.

e13049 Background: The clinical behaviors of de novo metastatic breast cancer (dnMBC) differ according to hormone receptor (HR) status, but an association between estrogen receptor (ER) expression and clinical outcomes has not been established. Our study aimed to investigate the influence of different ER status on survival outcomes for patients with dnMBC. This study was the Medical Science Research Project of Hebei Province and approved by the Ethics Committee of the Ethics Committee of Cangzhou Clinical College of Integrated Traditional Chinese and Western Medicine of Hebei Medical University (NO. 2017-AF29-058). Methods: Using hospital-based database, we identified patients with dnMBC diagnosed between 2010 and 2017, retrospectively collecting demographic data, tumor characteristics, treatment types and survival data. According to ER status, the patients were divided into ER-negative and ER-positive groups, and the propensity score matching (PSM) was used to balance the mixed bias. Overall survival (OS) was estimated by Kaplan-Meier method and log rank test. Univariable and multivariable analyses were performed using the Cox proportional hazard model to identify statistically significant prognostic factors. Results: A total of 322 patients were included. The median age was 53 (21-82) years, and the median follow-up was 8.5 years. The median OS for the PSM cohort was 29 months (95% confidence interval [CI], 26-35 months), and 5-year OS was 18.9%. OS differed significantly between the two groups (p = 0.0012), with a median OS of 36 months (95% CI, 32-41 months) in ER-positive group and 24 months (95% CI, 19-27 months) in ER-negative group. Multivariate analysis showed that clinical T-stage and initial metastatic sites were independent prognostic factors for survival in patients focused on ER status, with Ki-67 index, endocrine therapy and radiotherapy added to the ER-positive group, while initial and advanced chemotherapy added to the ER-negative group. Conclusions: For patients with dnMBC, there were substantial differences in OS according to ER status. ER-negative subtype is clinically different from ER-positive subtype, showing unfavorable tumor characteristics and poorer OS. We identified several prognostic factors that could guide therapy selection in this patient population. Clinical trial information: 20210957 .

Histogram analysis of multi-model high-resolution diffusion-weighted MRI in breast cancer: correlations with molecular prognostic factors and subtypes.

To investigate the correlations between quantitative diffusion parameters and prognostic factors and molecular subtypes of breast cancer, based on a single fast high-resolution diffusion-weighted imaging (DWI) sequence with mono-exponential (Mono), intravoxel incoherent motion (IVIM), diffusion kurtosis imaging (DKI) models. A total of 143 patients with histopathologically verified breast cancer were included in this retrospective study. The multi-model DWI-derived parameters were quantitatively measured, including Mono-ADC, IVIM-D, IVIM-D*, IVIM-f, DKI-Dapp, and DKI-Kapp. In addition, the morphologic characteristics of the lesions (shape, margin, and internal signal characteristics) were visually assessed on DWI images. Next, Kolmogorov-Smirnov test, Mann-Whitney U test, Spearman's rank correlation, logistic regression, receiver operating characteristic (ROC) curve, and Chi-squared test were utilized for statistical evaluations. The histogram metrics of Mono-ADC, IVIM-D, DKI-Dapp, and DKI-Kapp were significantly different between estrogen receptor (ER)-positive vs. ER-negative groups, progesterone receptor (PR)-positive vs. PR-negative groups, Luminal vs. non-Luminal subtypes, and human epidermal receptor factor-2 (HER2)-positive vs. non-HER2-positive subtypes. The histogram metrics of Mono-ADC, DKI-Dapp, and DKI-Kapp were also significantly different between triple-negative (TN) vs. non-TN subtypes. The ROC analysis revealed that the area under the curve considerably improved when the three diffusion models were combined compared with every single model, except for distinguishing lymph node metastasis (LNM) status. For the morphologic characteristics of the tumor, the margin showed substantial differences between ER-positive and ER-negative groups. Quantitative multi-model analysis of DWI showed improved diagnostic performance for determining the prognostic factors and molecular subtypes of breast lesions. The morphologic characteristics obtained from high-resolution DWI can be identifying ER statuses of breast cancer.

Characteristics and prognosis of young breast cancer patients with low expression of estrogen receptor

To evaluate the clinical characteristics and prognosis of young breast cancer patients with an ER of 1%–10%. Breast cancer patients aged ≤35 years old were selected and classified into three groups, ER-negative group, ER-low positive group (ER positivity: 1%–10%) and ER-high positive group (ER positivity: ≥10%), to compare their clinicopathological characteristics and prognosis. Of the 1387 patients assessed, 30.4% were ER-negative, 4.3% were ER-low positive, and 65.3% were ER-high positive. There was no difference in age, Tumor Node Metastasis (TNM) stage, histological type, adjuvant chemotherapy and adjuvant radiation therapy among the three groups (p &gt; 0.05). A higher histological grade and greater Human Epidermal Growth Factor Receptor-2 (HER-2) positivity were observed in the ER-low positive group than in the ER-high positive group (p &lt; 0.001). The number of patients with Progesterone Receptor (PR) negative in the ER-low positive group was between the other two groups. The recurrence rate of breast cancer in the ER-low positive group was 27.1%, which was similar to that of the ER-negative group (28%; p &gt; 0.05) but higher than the ER-high positive group (21.4%; p = 0.03). After a median follow-up of 74 months, the ER-high positive group had the longest Disease Free Survival (DFS) compared with the ER-negative group (p &lt; 0.0001) and ER-low positive group (p &lt; 0.05), while there was no significant difference in DFS between the latter two groups (p = 0.73). Similarly, the ER-high positive group had the longest Overall Survival (OS) than the ER-negative group (p &lt; 0.0001) and the ER-low positive group (p &lt; 0.05), while there was no statistical difference in OS between the latter two groups (p = 0.77). After endocrine therapy, no improvement in DFS (p = 0.71) and OS (p = 0.54) was observed in the ER-low positive group. In young breast cancer patients, the clinicopathological characteristics of the ER-low positive group were different from the ER-high positive group but were more similar to the ER-negative group. The DFS and OS were shorter than the ER-high group, and despite receiving endocrine therapy, DFS and OS of the ER-low positive group were not significantly prolonged.

Dual function miR-205 is positively associated with ER and negatively with five-year survival in breast cancer patients

BackgroundPrecise molecular characterization of breast cancer, especially triple negative (TNBC) as the most lethal subtype, is needed to stratify patients for the individual treatment approach. MicroRNA-205 (miR-205) has tumor-suppressive and oncogenic functions across different cancers. Therefore, miR-205 might have a different role in TNBC and estrogen receptor (ER) positive BC. Our aim was to investigate how miR-205 expression is associated with ER/progesteron receptor status, clinical parameters, pathohistological characteristics of BC, and survival of patients MethodsWe determined miR-205 relative expressions in 73 primary breast tumors (50 TNBC and 23 ER+) by quantitative Real-time polymerase chain reaction (qPCR) and compared it to clinicopathological characteristics and outcome. ResultsThe highest levels of miR-205 were in the ER+ /PR+ group, and the lowest in the TNBC group (p = 0.009). Significantly higher levels of miR-205 were also observed in the ER+ compared with the ER-negative group, regardless of the PR status (p = 0.002). Low miR-205 expression level was associated with prognostic stage III in TNBC samples (p = 0.049). Patients who received adjuvant chemotherapy had significantly lower levels of miR-205 (p = 0.016). Patients who received hormone therapy had significantly higher levels of miR-205 (p = 0.007). The low-miR-205 patients had significantly higher 5-year survival rates (p = 0.041). ConclusionThe expression of miR-205 in BC is subtype-specific and high expression is associated with the ER+ tumors. The miR-205 expression might be a useful marker of TNBC progression. High miR-205 expression had a detrimental effect on BC patient outcome. Our results indicate that miR-205 might be utilized in clinical practice as a biomarker and an adjunct parameter for the selection of the most effective therapeutic modality.

Combined 18F-FDG and 18F-Alfatide II PET May Predict Luminal B (HER2 Negative) Subtype and Nonluminal Subtype of Invasive Breast Cancer.

Noninvasive PET molecular imaging using radiopharmaceuticals is important to classify breast cancer in the clinic. The aim of this study was to investigate the combination of 18F-FDG and 18F-Alfatide II for predicting molecular subtypes of invasive breast cancer. Forty-four female patients with clinically suspected breast cancer were recruited and underwent 18F-FDG and 18F-Alfatide II PET/CT within a week. Tracer uptake in breast lesions was assessed using the maximum standardized uptake value (SUVmax), mean standardized uptake value (SUVmean), and SUVmax ratio of 18F-FDG to 18F-Alfatide II (FAR). Invasive breast cancer lesions were further classified as luminal A subtype, luminal B subtype, human epidermal growth factor receptor-2 (HER2) overexpressing subtype, and triple negative subtype according to the expression of the estrogen receptor (ER), progesterone receptor (PR), HER2, and Ki-67. Among 44 patients, 35 patients were pathologically diagnosed with invasive breast cancer. The SUVmax and SUVmean of 18F-FDG were significantly higher in the ER-negative group than those in the ER-positive group, as well as in the PR-negative group than those in the PR-positive group. However, the SUVmax and SUVmean of 18F-Alfatide II were higher in the ER-positive group and the PR-positive group. By combining 18F-FDG and 18F-Alfatide II, the FAR was lower in the ER-positive group and the PR-positive group. The HER2 overexpressing subtype showed the highest SUVmax and SUVmean for 18F-FDG while the luminal B (HER2 negative) subtype revealed the lowest values. The luminal B (HER2 negative) subtype showed the highest 18F-Alfatide II SUVmax, while the triple negative subtype showed the lowest 18F-Alfatide II SUVmax. The FAR was the lowest in the luminal B (HER2 negative) subtype and much higher in the HER2 overexpressing and triple negative subtypes. FAR less than 1 predicted the luminal B (HER2 negative) subtype with high specificity (93.1%) and NPV (90%). FAR greater than 3 predicted the HER2 overexpressing subtype and triple negative subtype (namely, the nonluminal subtype) with very high specificity (100%) and PPV (100%). In summary, FAR, the combined PET parameter of 18F-FDG and 18F-Alfatide II, can be used to predict molecular subtypes of invasive breast cancer, especially for the luminal B (HER2 negative) subtype and the nonluminal subtype.

UHRF1 modulates breast cancer cell growth via estrogen signaling.

The ubiquitination process, which involves that binding of an ubiquitin protein to certain substrates, regulates several human biological processes and human cancers. Several studies report that the abnormal expression of quite a few E3 ubiquitin ligases could play critical role in carcinogenic process and cancer progression. In our current study, we identify UHRF1 (Ubiquitin Like with PHD And Ring Finger Domain 1) is an important regulator for breast cancer growth. UHRF1 depletion significantly decreases breast cancer growth in vitro and in vivo. Clinical data analysis reveals that UHRF1 is dramatically elevated in breast cancer, compared to normal breast tissue. UHRF1 correlates with poor survival in luminal type of breast cancer patients, but not in ER-negative groups. The molecular biological studies show that UHRF1 localizes in the nuclear and interact with ERα via its SRA domain, which subsequently inhibits K48-linked ubiquitination of ERα and enhances ERα stability. Our study provides a novel function of UHRF1 in regulation estrogen signaling in breast cancer and a promising target for breast cancer therapeutics.

Effector memory cytotoxic CD3+/CD8+/CD45RO+ T cells are predictive of good survival and a lower risk of recurrence in triple-negative breast cancer

Triple-negative breast cancer (TNBC) with high tumour-infiltrating lymphocytes (TILs) has been associated with a promising prognosis. To better understand the prognostic value of immune cell subtypes in TNBC, we characterised TILs and the interaction between tumour cells and immune cell subtypes. A total of 145 breast cancer tissues were stained by multiplex immunofluorescence (mIF), including panel 1 (PD-L1, PD-1, CD3, CD8, CD68 and CK) and panel 2 (Foxp3, Granzyme B, CD45RO, CD3, CD8 and CK). Phenotypes were analysed and quantified by pathologists using InForm software. We found that in the ER-negative (ER <1% and HER2-negative) group and the ER/PR-low positive (ER 1–9% and HER2-negative) group, 11.2% and 7.1% of patients were PD-L1+ by the tumour cell score, 29.0% and 28.6% were PD-L1+ by the modified immune cell score and 30.8% and 32.1% were PD-L1+ by the combined positive score. We combined ER-negative and ER/PR-low positive cases for the survival analysis since a 10% cut-off is often used in clinical practice for therapeutic purposes. The densities of PD-L1+ tumour cells (HR: 0.366, 95% CI: 0.138–0.970; p = 0.043) within the tumour compartment and CD3+ immune cells in the total area (tumour and stromal compartments combined) (HR: 0.213, 95% CI: 0.070–0.642; p = 0.006) were favourable prognostic biomarkers for overall survival (OS) in TNBC. The density of effector/memory cytotoxic T cells (CD3+CD8+CD45RO+) in the tumour compartment was an independent prognostic biomarker for OS (HR: 0.232, 95% CI: 0.086–0.628; p = 0.004) and DFS (HR: 0.183, 95% CI: 0.1301–0.744; p = 0.009) in TNBC. Interestingly, spatial data suggested that patients with a higher density of PD-L1+ tumour cells had shorter cell-cell distances from tumour cells to cytotoxic T cells (p < 0.01). In conclusion, we found that phenotyping tumour immune cells by mIF is highly informative in understanding the immune microenvironment in TNBC. PD-L1+ tumour cells, total T cells and effector/memory cytotoxic T cells are promising prognostic biomarkers in TNBC.

Abstract P5-06-16: Radiosensitivity and immune cell infiltration signature predict clinical outcome of patients in the molecular taxonomy of breast cancer international consortium (METABRIC) study cohort

Abstract Purpose/Objectives There are growing evidences that tumor radiosensitivity is associated with immune activation in solid tumors. However, previous studies have not examined the relationship between patterns of immune infiltration and radiation response according to diverse molecular subtypes of breast cancer. This study evaluated radiosensitivity and immune infiltration signature to define a group of patients would benefit most from radiation therapy. Materials/Methods We performed integrative analyses of the clinical, genomic, transciptomc, and immunogenomic data to characterize the molecular features associated with radiosensitivity. We analyzed 1,903 data sets of METABRIC Study cohort using the radiosensitivity index (RSI), CIBERSORT and xCell, gene expression deconvolution algorithm which estimates the immune composition of tumor samples. According to RSI cut-point (0.3745), the patients were grouped into radiosensitive (RS, RSI-low tumors, n=1,082) versus radioresistant (RR, RSI-high tumors, n=821). Next, patients were divided into two groups accoring to immune and stromal scores, immune infiltrated (IF, n= 650) and immune excluded (IE, n=1253) subgroups. and performed integrative analyses of the clinical, genomic, transciptomc, and immunogenomic data to characterize the molecular features associated with radiosensitivity. Results RS group showed improved prognosis compared to RR group, and IF group had better survival than IE group, however, these tendency was only significant in ER negative group. Radiosensitivity was significantly associated with activation of antitumor immunity. In contrast, radio-resistance was associated with metastatic properties, such as epithelial mesenchymal transition and angiogenesis. Differentially expressed gene analysis revealed that ER signaling pathway is correlated with suppression of antitumor immunity. Higher ER was associated with higher fraction of M2 macrophage and lower PD-1 expression. Integration of immune signature and radiosensitivity index further stratified patients into four subgroups. In ER-negative disease, IF and RS group were associated with the best prognosis, whereas in ER-positive disease, immune signature and radiation response have no prognostic significance. On multivariable cox regression, both integrated immune infiltration and radiosensitivity signature was independent predictors of relapse free survival (HR 1.12, 95% CI 1.06-1.19, p &amp;lt; 0.001) and overall survival (HR 1.06, 95% CI 1.01-1.11, p = 0.028). Conclusion Taken together, this results suggested that tumor radiosensitivity was associated with activation of antitumor immunity and led to better relapse free survival particularly in ER negative group. Citation Format: Byung-Hee Kang. Radiosensitivity and immune cell infiltration signature predict clinical outcome of patients in the molecular taxonomy of breast cancer international consortium (METABRIC) study cohort [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-06-16.

Abstract P4-05-11: The clinical importance of the new category, ER low-positive in the ER expression HER2 negative early breast cancer

Abstract &amp;lt;Background&amp;gt;In the latest updated ASCO/CAP guideline, a new category, ER low-positive was introduced, which are determined by the 1% to10 % of positive staining of the tumor nuclei for ER. However clinical meaning of ER-low positive was unclear compared with ER- positive or ER-negative in HER2 negative breast cancer. &amp;lt;Purpose&amp;gt; In order to clarify the clinicopathological features of ER-low positive breast cancer, we compared the clinicopathological factors and prognosis among three groups, ER negative, ER-low positive, and ER-positive groups in the patients with HER2 negative breast cancer. &amp;lt;Patients and Method&amp;gt; A total of 1,882 patients with HER2 negative breast cancer who underwent surgery between 2001 and 2018 were included in this study. We divided them into three groups, ER negative (&amp;lt;1%), ER-low positive (1-10%), and ER positive (10%≦). The relationships between ER status and clinicopathological characteristics and prognosis were evaluated. &amp;lt;Result&amp;gt;According to ER status, 553(29.4%), 183(9.7%), 1146(60.9%) patients were divided into ER negative, low-positive and positive groups. Compared with ER-positive group, patients in the ER negative and low-positive groups were of higher age (p&amp;lt;.0001), and those tumors were significantly associated with the larger tumor size (p&amp;lt;.0001), higher histological grade (p&amp;lt;.0001) and more administration of adjuvant chemotherapy (p&amp;lt;0.0001). Most of the patients with ER low-positive and ER-positive tumors had received the adjuvant endocrine therapy. The prognosis of the patients with ER low-positive tumors were poorer than that of those with ER positive tumors in terms of relapse-free survival (RFS p&amp;lt;.0001), distant metastasis-free survival (DMFS p&amp;lt;.0001) and overall survival (OS p&amp;lt;.0001).In the node negative patients, the prognosis of the ER-negative and ER low-positive groups were equivalent, and poorer than that of the ER-positive group (RFS; p=0.0068, DMFS; p=0.0013, OS: p=0.0032). In addition, in pStage III patients, prognosis of the ER-negative and ER low-positive groups were also equivalent, and poorer than that of the ER-positive group (RFS; p=0.008, DMFS; p=0.0021, OS: p=0.0010). &amp;lt;Conclusion&amp;gt; This study showed that the prognosis of the patients with ER negative and ER low-positive were similar, and poorer than that of those with ER positive tumors. Therefore, the new category of ER-low positive is clinically especially important in order to determine the appropriate adjuvant therapy. Citation Format: Tajiri Wakako. The clinical importance of the new category, ER low-positive in the ER expression HER2 negative early breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-05-11.

Combination of Endogenous Estradiol and Adipokine Leptin in Breast Cancer Risk and Prognosis Assessment in Postmenopausal Chinese Women

ObjectiveOur study aims to clarify the role of estradiol and leptin in breast cancer risk and prognostic assessment in postmenopausal Chinese women.DesignThe serum circulating estradiol and leptin level was detected by ELISA. Then the correlation between estradiol, leptin level, and clinical characteristics was analyzed using Fisher’s exact test. Next, the Kaplan-Meier model was used to analyze the association between estradiol, leptin, and prognosis of postmenopausal breast cancer patients in our cohort and the TCGA dataset.SettingThe study was conducted at the National Cancer Center, Cancer Hospital, Chinese Academy of Medical Sciences, and Peking Union Medical College.PatientsA total of 182 postmenopausal breast cancer patients and 111 healthy subjects from January 2010 to August 2010 were included in the analysis. Another 702 cases with breast cancer were retrieved from The Cancer Genome Atlas (TCGA) database for subsequent analysis.Main Outcome MeasureSerum circulating estradiol and leptin level.ResultsThe level of estradiol was significantly higher (P<0.001) but the level of leptin had no significant difference (P = 0.764) in postmenopausal breast cancer patients compared with healthy subjects. The level of estradiol and leptin was not significantly different between estrogen receptor (ER) positive and ER-negative groups (P>0.05). Estradiol was significantly correlated with tumor T stage (P = 0.002) and leptin was significantly associated with perineural invasion (P = 0.014). In addition, the disease-free survival of patients with a high level of estradiol was significantly shorter (P = 0.025) but leptin tended to be a protective factor for overall survival in TCGA analysis (P = 0.038).ConclusionCirculating estradiol and leptin played important roles in the risk of postmenopausal breast cancer even in low-estrogen nations with an independent expression of ER status. High circulating estradiol was a poor prognostic factor and leptin may be a protection signal in Chinese postmenopausal patients with breast cancer.

Breast Imaging Findings of Microcalcifications in Ductal Carcinoma in Situ and Their Correlations with Pathological and Biological Features

Background: Mammography (MMG) is the primary screening tool for breast cancer, as microcalcifications are the most common MMG finding in ductal carcinoma in situ (DCIS). The use of high-frequency transducers facilitates the visualization of calcifications on ultrasonography (USG), especially in patients with dense breasts and cancer symptoms. Although a correlation has been reported between the imaging features of DCIS and pathological features, few studies have focused on multiple imaging modalities. Objectives: To evaluate the correlation of DCIS microcalcifications in breast imaging with pathological and biological features. Patients and Methods: The MMG and USG findings of 125 lesions detected in 123 patients, diagnosed with pure DCIS, were retrospectively reviewed according to the breast imaging-reporting and data system (BI-RADS). The USG and comparable MMG findings of microcalcifications were divided into three groups: group 1 (MMG negative, USG negative), group 2 (MMG positive, USG negative), and group 3 (MMG positive, USG positive). The pathological findings (nuclear grade and comedo necrosis) and biological features [estrogen (ER) positive group, human epidermal growth factor receptor 2 (HER2) positive group, triple negative group, and Ki-67 index] were compared with the MMG and USG features using Chi-square test. Results: Microcalcifications were observed on MMG in 83 (66.4%) DCIS lesions. Positive microcalcifications on MMG were significantly associated with a high nuclear grade (P = 0.001) and comedo necrosis (P = 0.001). Positive microcalcifications on MMG were significantly associated with ER negativity (P = 0.023), HER2 positivity (P = 0.002), and increased Ki-67 index (P = 0.001). There were 62 lesions (49.6%) without microcalcifications on USG (group 1 and group 2), while there were 63 (50.4%) lesions with microcalcifications on USG (group 3). Positive microcalcifications on MMG were significantly associated with ER-negative group (P = 0.023), HER2-positive group (P = 0.002), and increased Ki 67 index (P = 0.001). Conclusion: Based on the present results, DCIS microcalcifications detected via imaging were significantly associated with poor prognostic pathological factors, such as a high nuclear grade and comedo necrosis, as well as poor prognostic biological factors, including ER negativity, HER2 positive group, and a high Ki-67 index.

Radiosensitivity and Immune Cell Infiltration Signature Predict Clinical Outcome of Patients in the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) Study Cohort

There is growing evidence that tumor radiosensitivity is associated with immune activation in solid tumors. However, previous studies have not examined the relationship between patterns of immune infiltration and radiation response according to diverse molecular subtypes of breast cancer. This study evaluated radiosensitivity and immune infiltration signature to define a group of patients would benefit most from radiation therapy.We performed integrative analyses of the clinical, genomic, transcriptomic, and immunogenomic data to characterize the molecular features associated with radiosensitivity. We analyzed 1,903 data sets of METABRIC Study cohort using the radiosensitivity index (RSI), CIBERSORT and xCell, gene expression deconvolution algorithm which estimates the immune composition of tumor samples. According to RSI cut-point (0.3745), the patients were grouped into radiosensitive (RS, RSI-low tumors, n = 1,082) versus radioresistant (RR, RSI-high tumors, n = 821). Next, patients were divided into two groups according to immune and stromal scores, immune infiltrated (IF, n = 650) and immune excluded (IE, n = 1253) subgroups.RR group showed poorer relapse free survival compared to RS group in multivariate (HR 1.52, 95% CI 1.23-1.89, P < 0.001) analyses and this tendency was only significant in ER negative group. In multivariate logistic regression analysis, CD8+ T cells (OR = 1.21 95% CI 1.1-1.33 P < 0.001), gamma/delta T cells (OR = 1.3 95% CI 1.19-1.43 P < 0.001), dendritic cell resting (OR = 1.54 95% CI 1.22-1.95 P < 0.001), memory B cells (OR = 1.17 95% CI 1.03-1.33 P = 0.013), Pam50 luminal B1 subtype (OR = 1.59 95% CI 1.25-2.02 P < 0.001), and triple negative InClust 10 (OR = 1.67 95% CI 1.17-2.4 P = 0.005) were significantly enriched in RS group. In contrast, M0 macrophages (OR = 0.86 95% CI 0.78-0.94 P = 0.001), HER2 enriched integrative cluster subtype (InClust) 5 (OR = 0.43 95% CI 0.31-0.61 P < 0.001), ER positive InClust 4+ (OR = 0.5 95% CI 0.37-0.68 P < 0.001), and PIK3CA pathway alteration (OR = 0.75 95% CI 0.62-0.92 P = 0.005) were significantly associated with RR group.Taken together, these results suggested that tumor radiosensitivity was associated with activation of antitumor immunity and led to better relapse free survival particularly in ER negative group.

Response to neoadjuvant chemotherapy based on pathologic complete response in very young patients with ER-positive breast cancer: a large, multicenter, observational study

BackgroundIn estrogen receptor (ER)-positive breast cancer (BC), young age is associated with poor prognosis. While very young patients respond better to chemotherapy, chemotherapy is less effective in ER-positive tumors than in ER-negative tumors. The authors tried to evaluate chemotherapy response of very young patients with ER-positive BC by pathologic complete response (pCR) after neoadjuvant chemotherapy excluding the effect of endocrine treatment to the extent possible.MethodsWe collected individual patient data from 1992 to 2013 from the Korean Breast Cancer Society (KBCS). Total 1048 ER-positive and 797 ER-negative patients aged < 50 years who had been treated with neoadjuvant chemotherapy were included for analysis. We compared pCR rate between patients aged < 35 years with ER-positive tumors and the other groups.ResultsThe proportion of patients aged < 35 years was 14.0% of patients with ER-positive BC in this cohort of under 50 years old, and 16.8% of patients with ER-negative BC in this cohort of under 50 years old. Although most characteristics of tumors according to age were comparable, tumors with high Ki-67 expression were more common in patients aged < 35 years than in patients aged 35-49 years in both ER-positive and -negative group (P = 0.001). Breast conservation rates were not significantly different according to age (44.2% vs. 46.8% in ER-positive group, 55.2% vs. 48.0% in ER-negative group). pCR rate was not different according to age in ER-positive group (P = 0.71) but significantly better in patients aged < 35 years in ER-negative group (P = 0.009). After adjusting for confounding variables, young patients maintained the higher probability of pCR than older patients in ER-negative tumors. However, pCR rate did not differ according to age in ER-positive tumors. In multivariate analysis, young age (< 35 years) was correlated with poor overall survival (P = 0.003, HR = 1.98) and there was only one event in a few patients achieved pCR in ER-positive group.ConclusionsChemotherapy response based on pCR was not better in young patients (< 35 years) with ER-positive BC than in older premenopausal patients with non-metastatic ER-positive BC. Young age cannot be a predictive factor of response to neoadjuvant chemotherapy in ER-positive BC. Different biological characteristics such as high proliferative index should be considered.Trial registrationRetrospectively registered.

Age-Related Biology of Early-Stage Operable Breast Cancer and Its Impact on Clinical Outcome.

Simple SummaryBreast cancer incidence not only increases with advancing age but also changes its biology. This study was conducted to understand aging related change in the biological characteristics of breast cancer. The results highlighted that the change occurs in a gradual fashion, where 40 years and 70 years become the milestones for significant difference. Breast cancer in patients <40 years showed aggressive characteristics while at 70 years and above they are more indolent. The molecular pattern between 40 years and 70 years appears to be a transition from aggressive to less aggressive phenotypes. This change in the biology of the breast cancer significantly influences clinical outcome.As age advances, breast cancer (BC) tends to change its biological characteristics. This study aimed to explore the natural progression of such changes. The study included 2383 women with clinically T0-2N0-1M0 BC, managed by primary surgery and optimal adjuvant therapy in a dedicated BC facility. Tissue micro-arrays were constructed from their surgical specimens and indirect immunohistochemistry was used for analysis of a large panel (n = 16) of relevant biomarkers. There were significant changes in the pattern of expression of biomarkers related to luminal (oestrogen receptor (ER), progesterone receptors (PgR), human epidermal growth factor receptor (HER-2), E-cadherin, MUC1, bcl2 CK7/8, CK18 and bcl2) and basal (CK5/6, CK14, p53 and Ki67) phenotypes, lymph node stage, histological grade and pathological size when decade-wise comparison was made (p < 0.05). The ages of 40 years and 70 years appeared to be the milestones marking a change of the pattern. There were significantly higher metastasis free and breast cancer specific survival rates among older women with ER positive tumours while there was no significant difference in the ER negative group according to age. Biological characteristics of BC show a pattern of change with advancing age, where 40 years and 70 years appear as important milestones. The pattern suggests <40 years as the phase with aggressive phenotypes, >70 years as the less aggressive phase and 40–70 years being the transitional phase.

Characterization of estrogen receptor-low-positive breast cancer.

The biology of breast cancer with a low expression level (1-10%) of estrogen receptor (ER) remains a matter of confusion. The recent American Society of Oncology/College of American Pathologist Guidelines have recommended reporting such tumors as a new "ER-low-positive" category with a recommended comment to emphasize the possible overall benefit of endocrine therapies in these patients. The aim of the study was to analyze the clinicopathologic features and clinical outcomes of ER-low-positive breast cancers. We characterized the clinicopathologic features and survival outcomes of ER-low-positive breast cancers in our 4179 patients diagnosed from 1998 to 2018. The ER-positive, ER-low-positive, and ER-negative cases in our cohort were 2982 (71.4%), 97 (2.3%), and 1100 (26.3%), respectively. ER-low-positive tumors showed similar clinicopathologic characteristics yet significantly superior prognosis when compared to ER-negative tumors, while demonstrated largely overlapping survival outcomes with ER-positive tumors in the entire cohort. In the subcohort of tumors with a PR-positive phenotype, the prognosis of ER-low-positive tumors was intermediate between that of the ER-positive and ER-negative groups. ER-low-positive/PR-positive tumors had a significantly worse prognosis than ER-positive tumors, and a trend toward favorable survival outcomes when compared to ER-negative tumors, although no significant difference was identified for the latter. In contrast, the ER-positive and ER-low-positive groups showed similar survival outcomes in the subset of tumors with a PR-negative status, both being significantly better than ER-negative tumors. PR status as a surrogate marker of functional ER signaling provides critical information in this regard. These findings suggest that while ER-low-positive tumors are themselves heterogeneous, they often respond to endocrine treatment. Analysis of molecular signatures and standardization of therapeutic strategies are important to understand the biology of ER-low-positive tumors and to enable optimal treatment in the pursuit of individualized medicine.

Increased Expression of DNA2 Was Linked to Poor Prognosis in Breast Cancer.

DNA double-strand break (DSB) repaired by homologous recombination (HR) is an essential process for breast cancer cells to survive. DNA2 nuclease acts parallel to homologous recombination (HR). Here, we investigated the detailed clinical attribute of DNA2 in breast cancer and the role of DNA2 in breast cancer cells' growth. We found that elevated expression of DNA2 was obviously linked to poor prognosis in breast cancer. Further, DNA2 expression was increased in the ER-negative group, PR-negative group, HER2-positive group, and high-grade group via analyzing 2,509 breast cancers in “cBioportal” and 3,063 breast cancer data in “bc-GenExMiner.” Besides, the immunohistochemical staining in 26 breast cancer tissues also showed that elevated expression of DNA2 was correlated with ER-/PR-/HER+. To further detect the role of DNA2 in breast cancer cells, we took GESA, GO, and KEGG analyses and found that DNA2 was enriched in cell cycle and DNA replication pathways. Furthermore, silencing of DNA2 inhibited cell growth in T47D and MD-MB-231 breast cancer cells and suppressed tumor growth in vivo, indicating DNA2 functioned importantly in breast cancer progression and maybe a potential prognostic marker in breast cancer. Our research reveals that DNA2 is a biomarker for diagnosis and prognosis in breast cancer from multiple perspectives and gives a new clue for further preclinical and clinical investigation.