Estrogen Receptor Negative Research Articles

Complete pathological response in patients with HER2 positive breast cancer treated with neoadjuvant therapy in Colombia

Breast cancer is the most common type of cancer and the leading cause of death by cancer in women in Colombia. Approximately 15 to 20% of breast cancers overexpress HER2. To analyze the relationship between multiple clinical and histological variables and pathological complete response in patients with HER2-positive breast cancer undergoing neoadjuvant therapy in a specialized cancer center in Colombia. We performed a retrospective analysis of non-metastatic HER2-positive breast cancer patients who received neoadjuvant therapy between 2007 and 2020 at the Instituto de Cancerología Las Americas Auna (Medellín, Colombia). Assessed parameters were tumor grade, proliferation index, estrogen receptor, progesterone receptor, HER2 status, type of neoadjuvant therapy, pathologic complete response rates, and overall survival. Variables associated with low pathologic complete response rates were tumor grades 1-2 (OR = 0.55; 95% CI = 0.37-0.81; p = 0.03), estrogen receptor positivity (OR =0.65; 95%; CI = 0.43-0.97; p=0.04), and progesterone receptor positivity (OR = 0.44; 95% CI = 0.29-0.65; p = 0.0001). HER2 strong positivity (score 3+) was associated with high pathological complete response rates (OR = 3.3; 95% CI = 1.3-8.35; p=0.013). Five-year overall survival was 91.5% (95% CI = 82.6-95.9) in patients with pathological complete response and 73.6% (95% CI = 66.4-79.6) in patients who did not achieve pathological complete response (p = 0.001). Additionally, the pathological complete response rate was three times higher in patients receiving combined neoadjuvant chemotherapy with anti-HER2 therapy than in those with chemotherapy alone (48% versus 16%). In patients with HER2-positive breast cancer, tumor grade 3, estrogen receptor negativity, progesterone receptor negativity, strong HER2 positivity (score 3+), and the use of the neoadjuvant trastuzumab are associated with higher pathological complete response rates.

Effectiveness and Toxicity of Five Fraction Prone Accelerated Partial Breast Irradiation

Accelerated partial breast irradiation (APBI) after breast conserving therapy (BCT) is increasingly used to treat women with early stage breast cancer. Our institution was an early adopter of 5-fraction ABPI and delivers APBI primarily to patients in the prone position. This study reports long term oncologic and cosmetic outcome in a large cohort of women treated with 5-fraction external beam APBI. We included patients receiving APBI 600 cGy × 5 fx delivered every other day or every day between 2010 and 2022. Late toxicities and clinician and patient rated cosmesis were evaluated for patients with > 6 month follow up. Univariate and multivariate logistic regression models were used to identify clinical and dosimetric factors associated with development of acute and late toxicities, clinician and patient rated cosmesis. All statistical tests were two-sided, and the null hypothesis was rejected for p<0.05. Kaplan Meier methodology was used to calculate overall survival (OS), disease-free survival (DFS) and locoregional recurrence-free survival (LR-RFS). Four hundred and forty-two patients received APBI either daily (56%) or every other day (44%). Most of the patients (92%) were treated in the prone position. Average mean heart dose was 23 cGy for left-sided and 11 cGy for right-sided breast cancers. Ipsilateral lung V30% ≤ 30%. At a median follow up of 48 months (range: 5.96 - 155 months), 12 (2.7%) patients developed a local recurrence, 14 (3.2%) patients developed a contralateral breast primary, 10 patients (2.3%) developed a distant metastasis and one patient (0.2%) developed a local recurrence followed by a distant metastasis 1 month later. Out of 258 patients with > 6 month follow up, rates of late grade 1-2 telangiectasia, fibrosis, edema, atrophy and hyperpigmentation were 4%, 18%, 1%, 19% and 7% respectively. There was only one late grade 3 event in a patient who developed significant breast atrophy. The rate of good-excellent physician and patient rated cosmesis was 95% and 89% respectively (N = 256 at median follow up of 80 months). On multivariate logistic regression, patients who did not receive any adjuvant endocrine or chemotherapy were at increased risk of developing a local recurrence. Patients with PR negative disease were at increased risk of distant metastasis. Patient who experienced any grade of acute dermatitis during treatment were at increased risk of any high grade (grade ≥ 2) late adverse event and worse physician rated cosmesis. Daily or every other day treatment did not correlate with worse toxicity or clinical outcomes. Plastic surgery involvement, LVI, EIC, lobular histology, and ER negativity did not correlate with an increased risk of recurrence. Five-year LR-RFS, DFS and OS were 98%, 92.5% and 98.6% respectively. Five- fraction APBI delivered primarily in the prone position either daily or every other day was effective with low rates of local recurrence, minimal toxicity and excellent cosmetic scores at long term follow up.

Is Ki-67 Really Useful as a Predictor for Response to Neoadjuvant Chemotherapy in Locally Advanced Breast Cancer?

Neoadjuvant chemotherapy (NACT) is routinely offered to operable locally advanced breast cancer (LABC) patients desirous of breast conservation surgery and inoperable LABC patients. Pathological complete response (pCR) following chemotherapy is recognized as a surrogate for survival outcomes in high grade tumour subtypes. Many biological and tumor characters have been shown to predict pCR. The current study was performed with the aim of investigating the ability of Ki-67 in predicting pCR with NACT in breast cancer patients. A total of 105 patients with locally advanced breast cancer who completed NACT followed by surgery were included in this study from January 2020 till December 2022. Patients with advanced metastatic breast carcinoma, who did not give consent for NACT, who did not complete NACT and who did not undergo surgery were excluded. All patients were assessed for Ki-67 score on core-needle biopsy samples and response rate was assessed clinically and by histopathological examination of resected specimen. Quantitative variables were compared using unpaired t-test or Mann-Whitney 'U' test and for categorical variables Chi-square or Fisher's exact test were used. Receiver operating characteristic (ROC) curve analysis was performed to assess the predictive potential of Ki-67 expression levels in predicting pCR. To identify the predictive factors associated with pCR, univariate analysis was performed. The P value < 0.05 was considered as statistically significant. Mean age was 51.57 ± 10.8years. 51 patients achieved clinical complete response (cCR) and 33 achieved pCR after NACT. Mean Ki-67 index in overall study population, in pCR group and no pCR group was 46.44 ± 22.92%, 51.60 ± 22.3% and 44.06 ± 22.7%, respectively. On univariate analysis, ER negativity, PR negativity and Her 2neu positivity were found predictive of pCR. On subgroup analysis, TNBC and Her 2neu positive sub groups were associated with higher cCR and pCR rate. We found no significant association between Ki-67 and pCR. This result may be confounded by the fact that a significant duration of the study was in the COVID-19 pandemic. Validation of this data is required in a large prospective study.

Prognostic and Predictive Significance of Tumor Immune Microenvironment in Breast Ductal Carcinoma In Situ

This study aims to identify the role of different subtypes of tumor infiltrating lymphocytes (TILs) in ductal carcinoma in situ (DCIS) in predicting risk of recurrence and benefit of whole breast irradiation (WBI). Immunohistochemical stain for CD3, CD4, CD8, FOXP3 and CD20 were carried in a well characterized DCIS cohort who received breast-conserving surgery (BCS) from Jan 2009 to Dec 2018. All the TILs subtypes were evaluated by the average numbers of touching-TILs which defined as TILs touching or within one lymphocyte cell thickness from the malignant ducts' basement membrane. The optimal cut-off values of TILs subtypes were selected by the X-tile. In total, 167 patients were enrolled in this analysis with 114 patients received WBI. After a median follow-up of 67 months, 15 IBTR events occurred with 6 invasive-IBTRs. Nine out of 15 IBTRs occurred outside of the original quadrant (elsewhere failure event, EFE). CD3+ lymphocytes were the predominant cell subtype while Treg showed the lowest levels. High abundance of TILs subtypes was associated with high tumor grade, presence of microinvasion, high Ki67 index, ER negativity and HER2 positivity. For various TILs subtypes, the multivariate analyses showed that dense CD4+ TILs (HR = 9.84, 95% CI 2.43-39.91, p<0.01) and dense Treg (HR = 4.22, 95% CI 1.24-14.36, p = 0.02) were independent prognostic factors for higher IBTR. As the infiltration of TIL subsets was correlated with one another, we also analyzed the relationship between IBTR and the ratios of different TILs subtypes. By adjusted by clinicopathological parameters, high ratios of CD4+/CD8+, Treg/CD4+ and Treg/CD8+ were found to be independent prognostic factors for higher IBTR (HR = 11.31, 95% CI 3.14-40.76, p<0.01; HR = 3.09, 95% CI 1.05-9.11, p = 0.04; HR = 7.14, 95% CI 1.98-25.73, p<0.01). Consistent with the results of IBTR, the 5-y rate of invasive-IBTR and EFE was both significantly associated with the high CD4+/CD8+, Treg/CD4+ and Treg/CD8+ TILs ratios (all p<0.01). WBI reduced the rate of 5y-IBTR risk from 8.4% to 1.3% (p = 0.02) in the low Treg/CD8+ group, but there was no benefit of WBI in the high group. With respect to EFE, WBI significantly reduced the rate from 2.8% to 0.0% (p = 0.03) in the low Treg/CD8+ group while not in the high group. The benefits of WBI in reducing IBTR and EFE were not significant difference between different CD4+/CD8+ and Treg/CD4+ groups. Assessment of overall TILs provides a tool for comprehensive evaluation of the DCIS immune microenvironment. Patients with pro-tumoral immune infiltrate (high Treg, high ratios of CD4+/CD8+, Treg/CD4+ and Treg/CD8+) in tumor microenvironment show an increased risk of IBTR and less benefit from breast radiotherapy.

Survival Analysis of Breast Cancer in Patients with HER2/neu+, ER-, PR-: a Retrospective Cohort Study

Aim: We aimed to study the characteristics of the breast cancer patients with positive HER2/neu+, ER- and PR- receptors. Methods: 119 patients with breast cancer were included in this retrospective cohort study from 2006 to 2016. The overall and disease-free survival were evaluated. Results: Most prevalent type of tumor was IDC, grade III cancer and stage II. Recurrence/metastasis occurred in 18.49%. Most common sites of metastasis were lungs and liver. Total mortality rate was 10.92%. Overall and disease-free survival times were 40.04 and 22.29 months, respectively. The median survival time was about 118 months. Conclusion: Breast cancers with positive HER2/neu and negative estrogen and progesterone receptors had low overall and disease-free survival rates compared with ER+/PR+/HER2- tumors.

The prognostic significance of skin involvement in breast cancer patients with chest wall recurrence

Purpose To assess the prognostic significance of skin involvement in breast cancer patients with chest wall recurrence (CWR). Methods We retrospectively analyzed the clinicopathological data of breast cancer patients with CWR who were diagnosed pathologically between January 2000 and April 2020. Disease-free survival (DFS) was the time from radical resection for CWR to disease recurrence. Progression-free survival (PFS) was defined as the time from the diagnosis of locally unresectable CWR to the first sign of disease progression. Persistent chest wall progression was defined as three consecutive chest wall progressions with no distant organ involvement. Results A total of 476 patients with CWR were included in this study. Skin involvement was confirmed in 345 patients. Skin involvement was significantly correlated with a high T stage (p = 0.003), more positive nodes at initial examination (p < 0.001) and lymphovascular invasion (p < 0.001). Kaplan–Meier analysis showed that skin involvement was a predictor of shorter DFS (p < 0.001), including both local disease progression (p < 0.001) and distant disease progression (p = 0.022). Multivariate analysis showed that skin involvement was an independent biomarker for DFS (p = 0.043). Patients with skin involvement were more likely to experience persistent chest wall progression (p = 0.040). After eliminating the potential deviation caused by an insufficient follow-up time, persistent chest wall progression was more likely to be associated with a high N stage (p = 0.002), negative progesterone receptor (PR; p = 0.001) and positive human epidermal growth factor receptor 2 (HER2; p = 0.046) of the primary site, and negative oestrogen receptor (ER; p = 0.027) and PR (p = 0.013) of the chest wall lesion and skin involvement (p = 0.020). Conclusion Skin involvement was a predictor of poor disease control in patients with CWR and was closely related to persistent chest wall progression. We stratified the prognosis of individualized treatment for breast cancer patients with CWR to provide new insights into the biological behaviours of the disease.

Immunohistochemical expression of Hairy and Enhancer of Split 1 (HES1) protein in breast carcinoma

Background Hairy and Enhancer of Split 1 (HES1) is a transcription factor involved in cell differentiation, proliferation, and in various cancer progression. Recently, HES1 was addressed as a potential biomarker that induces stem cell features in breast cancer and stimulates epithelial-mesenchymal transition in triple-negative breast cancer (TNBC). Aim This study aimed to assess HES1 immunohistochemical (IHC) expression in invasive breast carcinoma and correlate it with the clinicopathological variables and different molecular subtypes of the cases. Patients and methods IHC was conducted on 76 paraffin blocks with an anti-HES1 monoclonal antibody, and a final IHC score was calculated based on staining intensity and percentage of positively stained cells. A final IHC of greater than or equal to 4 was considered an HES1 positive expression. Results HES1 expression was positive in 18 (23.7%) cases and was significantly associated with breast cancer HER2-enriched molecular subtype, followed by triple-negative breast cancer (P&lt;0.05). HES1 expression was also significantly associated with estrogen and progesterone receptor negativity and HER2 overexpression (P&lt;0.05). Positive HES1 expression was associated with high tumor grade, advanced stage, high Ki-67 proliferation index, and presence of tumor vascular emboli, although it did not reach statistical significance (P&gt;0.05). Conclusion Our study demonstrated that the HES1 protein showed a significantly higher expression in nonluminal breast cancer subtypes. In addition, it could have a potential role in the growth and spread of breast cancer cells. These findings suggest that HES1 is a possible promising target for therapeutic interventions.

Enhanced confinement induced by pellet injection in the stellarator TJ-II

Enhanced confinement is observed in neutral beam injector (NBI)-heated hydrogen discharges made in the stellarator TJ-II after the injection of a single cryogenic fuel pellet into the plasma core. In addition to the expected increase in electron density, ne, in the core after pellet injection (PI), the plasma diamagnetic energy content is seen to rise, with respect to similar discharges without PI, by up to 40%. Furthermore, the energy confinement time, τEdiag, as determined using a diamagnetic loop, is enhanced when compared to predictions obtained using the International Stellarator Scaling law [H. Yamada et al., Nucl. Fusion 45, 1684 (2005)] and the triple product, ne · Ti · τEdiag, exhibits a clear bifurcation point toward an improved confinement branch as compared to the branch product predicted by this scaling law. In general, once such a pellet-induced enhanced confinement (PiEC) phase has been established, it is characterized by steepened radial density gradients, by more negative plasma potential in the core, more negative radial electric fields, Er, across a broad plasma region, as well as by reductions in density and plasma potential fluctuations in the density gradient region. In addition, experimental observations show increased peaking of core radiation losses, this pointing to edge/core plasma decoupling. In parallel, neoclassical simulations of reference and PiEC plasmas predict increased particle and energy confinement times during a PiEC phase together with a more negative Er profile. Qualitative rather than quantitative agreement with experimental parameters is found, indicating that turbulence seems to play a significant role here. In summary, single cryogenic pellet injection facilitates the achievement of an enhanced operational regime that was previously not observed in NBI-heated discharges of the TJ-II.

Prognostic implications of HER2 changes after neoadjuvant chemotherapy in patients with HER2-zero and HER2-low breast cancer

BackgroundThe transition of human epidermal growth factor receptor 2 (HER2) status after neoadjuvant chemotherapy (NAC) in HER2-low breast cancer has not been thoroughly evaluated. Here, we evaluated the HER2 transition among HER2-zero and HER2-low breast cancer cases post-NAC and its impact on clinical outcomes. MethodsWe included 1288 patients with HER2-low or zero breast cancer who underwent NAC and surgery between 2014 and 2018 and had paired pre- and post-therapeutic HER2 status results. ResultsAmong patients who were HER2-zero pre-NAC (n = 650), 68% and 29% were HER2-zero and HER2-low, respectively, post-NAC. Among patients who were HER2-low pre-NAC (n = 638), 32% of patients showed HER2 changes (low to zero), and 59% of patients had a constant HER2-low status post-NAC. Patients with constant HER2-low or transitions from HER2-low to zero had a higher proportion of hormone receptor positivity (84% and 79%) than those with changes from HER2-zero to low (77%) or with constant HER2-zero (56%), respectively. Multivariable logistic regression analysis revealed that patients with oestrogen receptor positivity had a higher probability of gaining HER2-low expression than those with oestrogen receptor negativity (odds ratio 2.48). No significant differences were observed in terms of overall survival or disease-free survival between patients with and without HER2-changes according to their hormone receptor status, except in the post-therapeutic HER2-low, hormone receptor-negativity subset. ConclusionTemporal heterogeneity of HER2-low expression is observed in substantial numbers of post-NAC breast cancer patients. Clinical outcomes show no significant associations, except in the post-therapeutic HER2-low, hormone receptor negativity subset. The prognostic implications of HER2 transition in HER2-low breast cancer require further investigation.

Negative estrogen receptor expression assessed by 18F-FES PET in metastatic breast cancer with ER-positive primary tumor.

1034 Background: The 18F-fluoroestradiol positron emission tomography/computed tomography (18F-FES PET/CT) technique provides a convenient method to evaluate the overall estrogen receptor (ER) expression in metastatic breast cancer (MBC) patients. There are long debates on the characteristics and treatment strategy of patients with positive primary ER lesion but negative ER expression in metastatic disease. 18F-FES PET offers an opportunity to answer this question. Methods: This study enrolled MBC patients with ER-positive primary tumor who received 18F-FES PET/CT in our center. Descriptive statistics were used in clinicopathologic characteristics and compared with Chi square test or t test. Progression‐free survival (PFS) was estimated by Kaplan-Meier method and compared by log-rank test. Results: 16.46% (52/314) patients with an ER-positive primary tumor had negative ER expression assessed by 18F-FES for MBC prior to receiving first-line systemic therapy. The rate of ER-negativity tested by 18F-FES was negatively co-related with levels of ER expression in early stage, with a positive to negative conversion in 2 of 3 (46.3%) ER-low (ie, ER 1%-9%) tumors, 7 of 10 (70.0%) ER-moderate (ie, ER 10%-49%) tumors, 16 of 114 (14.0%) ER-high (ie, ER 50%-95%) tumors, and 1 of 8 (12.5%) ER-very high (ie, ER &gt; 95%) tumors (p &lt; 0.001). Chemotherapy (83.33%, 40/48) was the most common treatment strategy afterward, among which capecitabine monotherapy (64.29%, 27/42) was a dominant alternative. PFS was significantly prolonged with capecitabine alone versus other chemotherapy (median PFS: 13.14 vs 6.21 months, p = 0.035). Conclusions: 18F-FES could identify negative conversion of ER in MBC which occurred frequently. Patients with lower ER expression in the primary lesion were more likely to have negative ER expression in the metastasis. In real-world clinical practice, chemotherapy was the primary choice by most physicians and capecitabine monotherapy had shown good efficacy.

Can Axillary Lymph Node Dissection be Omitted in Breast Cancer Patients with Metastatic Sentinel Lymph Nodes Undergoing Mastectomy? A Systematic Review and Meta-Analysis of Real-World Evidence.

The omission of axillary lymph node dissection (ALND) in patients with breast cancer who have metastatic sentinel lymph nodes (SLNs) undergoing mastectomy remains controversial. This meta-analysis explored the clinicopathological factors affecting the selection of ALND and the influences of ALND on survival outcomes in patients receiving mastectomy with positive SLNs. Eligible studies published prior to 31 December 2022 were selected by searching the Embase, Web of Science and PubMed databases. Pooled analyses were performed using the number of events for clinicopathological parameters and HRs with 95% CIs for survival outcomes including disease-free survival (DFS), overall survival (OS), distant recurrence-free survival (DRFS) and locoregional recurrence-free survival (LRFS). A total of 10 retrospective studies enrolling only breast cancer patients with limited SLN metastases (no more than 3 positive SLNs or micrometastatic SLNs) undergoing mastectomy were included. Performing ALND in mastectomy patients who had limited SLN metastases was significantly correlated with invasive ductal carcinomas, larger tumors, lymphovascular invasion, higher tumor grade, macrometastatic SLNs, more positive SLNs, extranodal extension, positive surgical margins, negative ER, administration of adjuvant chemotherapy and nonwhite race (P < 0.05). However, performing ALND did not result in significantly longer OS, DFS, LRFS or DRFS (P > 0.05) in these patients. The present meta-analysis indicated that ALND may be safely avoided in patients with breast cancer who had limited SLN metastases undergoing mastectomy. Further well-designed randomized clinical trials are warranted to validate our results.

Pattern of MUC6 expression across 119 different tumor types: A tissue microarray study on 15 412 tumors.

Mucin 6 (MUC6) is a secreted gel-forming mucin covering the surfaces of gastrointestinal and other tissues. Published work demonstrates that MUC6 can also be expressed in several cancer types and can aid in the distinction of different tumor entities. To systematically analyze MUC6 expression in normal and cancerous tissues, a tissue microarray containing 15 412 samples from 119 different tumor types and subtypes as well as 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. At least a weak MUC6 positivity was seen in 50 of 119 (42%) tumor entities. Thirty-three tumor entities included tumors with strong positivity. MUC6 immunostaining was most frequent in mucinous carcinomas of the breast (44%), adenocarcinomas of the stomach (30%-40%) and esophagus (35%), and neuroendocrine carcinomas of the colon. Strong MUC6 staining was linked to advanced pT stage (p = 0.0464), defective mismatch repair status and right-sided tumor location (p < 0.0001 each) in colorectal cancer, as well as to high tumor grade (p = 0.0291), nodal metastasis (p = 0.0485), erb-b2 receptor tyrosine kinase 2 positivity (p < 0.0001) and negative estrogen receptor (p = 0.0332)/progesterone receptor (p = 0.0257) status in breast carcinomas of no special type. The broad range of tumor types with MUC6 expression limits the utility of MUC6 immunohistochemistry for the distinction of different tumor types.

De-escalation of Sentinel Lymph Node Biopsy in Patients With Ductal Carcinoma In Situ.

Introduction Current guidelines recommend that sentinel lymph node biopsy (SLNB) be performed in patients with ductal carcinoma in situ (DCIS) undergoing mastectomy, in patients for whom the location of excision may compromise future SLNB, or if there is a high suspicion or risk of upstaging to invasive cancer on final pathology. Whether axillary surgery should be performed in patients with DCIS remains controversial. Our study aimed to examine the factors associated with the upgrade of DCIS to invasive cancer on final pathology and sentinel lymph node (SLN) metastases to evaluate whether axillary surgery may be safely omitted in DCIS. Methods Patients with a diagnosis of DCIS on core biopsy who underwent surgery with axillary staging between 2016 and 2022 were identified from our pathology database and retrospectively reviewed. Patients who underwent surgical management of DCIS without axillary staging and those treated for local recurrence were excluded. Results Out of 65 patients, 35.3% of patients were upstaged to the invasive disease on final pathology. 9.23% of cases had a positive SLNB. Predictive factors associated with upstaging to invasive cancer included palpable mass on clinical examination (P = 0.013), presence of a mass on preoperative imaging (P = 0.040), and estrogen receptor status (P = 0.036). Conclusion Our results support ongoing opportunities for the de-escalation of axillary surgery in patients with DCIS. In a subset of patients undergoing surgery for DCIS, SLNB may be omitted as the risk of upstaging to invasive cancer is low. Patients with a mass on clinical examination or imaging and negative estrogen receptor (ER) lesions have a higher risk of upstaging to invasive cancer, where a sentinel lymph node biopsy should be performed.

Positive surgical margins after breast-conserving surgery for ductal carcinoma in-situ: does histologic grade or estrogen receptor status matter?

DCIS has been shown to have a higher rate of positive margins following breast-conserving surgery (BCS) than invasive breast cancer. We aim to analyze certain factors of DCIS, specifically histologic grade and estrogen receptor (ER) status, in patients with positive surgical margins following BCS to determine if there is an association. A retrospective review of our institutional patient registry was performed to identify women with DCIS and microinvasive DCIS who underwent BCS by a single surgeon from 1999 to 2021. Demographics and clinicopathologic characteristics between patients with and without positive surgical margins were compared using chi-square or Student's t-test. We assessed factors associated with positive margins using univariate and multivariable logistic regression. Of the 615 patients evaluated, there was no significant difference in demographics between the patients with and without positive surgical margins. Increasing tumor size was an independent risk factor for margin positivity (P = < 0.001). On univariate analysis both high histologic grade (P = 0.009) and negative ER status (P = < 0.001) were significantly associated with positive surgical margins. However, when adjusted in multivariable analysis, only negative ER status remained significantly associated with margin positivity (OR = 0.39 [95% CI 0.20-0.77]; P = 0.006). The study confirms increased tumor size as a risk factor for positive surgical margins. We also demonstrated that ER negative DCIS was independently associated with a higher rate of positive margins after BCS. Given this information, we can modify our surgical approach to reduce rate of positive margins in patients with large-sized ER negative DCIS.

Autophagy in antitumor activity of aloin for breast cancer cells compared with doxorubicin

Breast cancer is the most commonly diagnosed cancer and is one of the leading causes of cancer mortality in women worldwide. Natural product compounds have attracted significant attention for their potent effects against human cancers. Aloin, a natural phytochemical anthraquinone glycoside extracted from Aloe sp., has been previously reported for its antitumor activity. Autophagy is a highly conserved process that mediates the degradation of dysfunctional cellular components, such as senescent proteins and organelles. In the present study, we verified the involvement of autophagy in tolerance to aloin, especially in breast cancer cells with negative estrogen receptors, and as an alternative pathway to promote cell death in cells expressing mutant p53 status, which often limits the efficacy and accounts for resistance to chemotherapy. We studied the effect of aloin on 2 types of breast cancer cell lines, estrogen receptor-positive (T47D) and triple negative (MDA-MB231), and compared to an anthraquinone analog, doxorubicin (Dox) as a reference compound. Aloin inhibited the cell growth of both T47D and MDA-MB231 cancer cells, in a time- and dose-dependent manner with a more pronounced effect in the 72 h exposure regimen, and in the ERα+ breast cell line. The autophagic activity of aloin was emphasized by the formation of autophagosomes and autolysophagosomes, as early and late autophagic compartments, respectively, as well as the accumulation of acidic vesicular organelles in the tumor cells. Also, upregulation in the protein expression of some marker genes of autophagy such as beclin 1 and LC3BII/LC3I, and conversely down-regulation in pmTOR and p62 was recorded. The results suggest that autophagy can be regarded as one of the mechanistic modes of aloin cytotoxicity in breast cancer cells that evade apoptosis through genetic mutations in p53.

Incidence, Clinicopathologic Features, HER2 Fluorescence In Situ Hybridization Profile, and Oncotype DX Results of Human Epidermal Growth Factor Receptor 2-Low Breast Cancers: Experience From a Single Academic Center

Human epidermal growth factor receptor 2 (HER2)-low breast cancer, defined by an immunohistochemical (IHC) score of 1+ or 2+ with negative in situ hybridization, is emerging as a predictive marker for the use of the antibody-drug conjugate. To understand how this category differs from HER2-zero cases, we investigated clinicopathological characteristics and HER2 fluorescence in situ hybridization results in a large cohort of 1309 continuous HER2-negative invasive breast carcinomas from 2018 to 2021 evaluated by the Food and Drug Administration-approved HER2 IHC test. Additionally, we compared Oncotype DX recurrence scores and HER2 mRNA expression between HER-low and HER2-zero cases in a separate cohort of 438 estrogen receptor-positive (ER+) early-stage breast carcinoma cases from 2014 to 2016. Based on the cohort from 2018 to 2021, the incidence of HER2-low breast cancers was approximately 54%. HER2-low cases had less frequent grade 3 morphology, less frequent triple-negative results, ER and progesterone receptor negativity, and a higher mean HER2 copy number and HER2/CEP17 ratio than HER2-zero cases (P < .0001). Among ER+ cases, HER2-low cases showed significantly less frequent Nottingham grade 3 tumors. In the cohort from 2014 to 2016, HER2-low cases showed significantly higher ER+ percentages, fewer progesterone receptor-negative cases, lower Oncotype DX recurrence scores, and higher HER2 mRNA expression scores than HER2-zero cases. In summary, this is the first study, to our knowledge, using a large cohort of continuous cases evaluated by the Food and Drug Administration-approved HER2 IHC companion diagnostic test for HER2-low expression and HER2 fluorescence in situ hybridization profile in a real-world setting. Although HER2-low cases showed a higher HER2 copy number, ratio, and mRNA level than HER2-zero cases statistically, such small differences are unlikely to be biologically or clinically meaningful. However, our study suggests that HER2-low/ER+ early-stage breast carcinoma may represent a less aggressive group of breast carcinoma, given its association with a lower Nottingham grade and Oncotype DX recurrence score.

Immunohistochemical expression of CD155 in invasive female breast carcinoma and its correlation with tumor infiltrating natural killer cells

BackgroundCD155 is an immune checkpoint protein that interacts with ligands on natural killer cells to regulate the tumor associated immunity. CD155 overexpression has been detected in many human cancer types. CD155 and its pathways are promising tumor immunotherapy targets. We aimed to evaluate the immunohistochemical expression of CD155 in invasive breast carcinomas and to correlate such expression with the pathological parameters of the tumors and also with natural killer - tumor infiltrating lymphocytes (NK-TILs) density in breast carcinomas tissue as highlighted by CD56 immunostaining. This study included 78 cases of breast carcinomas. Immunohistochemistry was performed using antibodies against CD155 which was detected on the tumor cells and CD56 as a marker for stromal NK cells.ResultsCD155 expression by the tumor cells was detected in 30.8% of the cases and correlated significantly with advanced prognostic stage, Estrogen receptor (ER) and Progesterone receptor (PR) negativity, high Ki-67 index and Human epidermal receptor 2 (HER2) enriched molecular subtype. High stromal TILs CD56 expression was detected in 28.2% of the cases and correlated significantly with high histologic grade, PR negativity, HER2 neu over-expression, high Ki-67 index, high stromal TILs and more aggressive molecular subtypes; triple negative breast cancer, HER2 enriched and Luminal B-HER2 positive. Finally, statistically significant direct correlation was detected between Tumor cells CD155 expression and high TILs CD56 expression.ConclusionsOur results support tumor cell CD155 expression and TILs CD56 expression in breast cancers that are high grade, TILs rich and hormone receptors negative, highlighting those cases as possible candidates for CD155 targeted therapy.

Abstract P3-04-10: The TILs-US score adding vascularity assessment based on ultrasonography for predicting tumor-infiltrating lymphocytes in human epidermal growth factor receptor 2-positive and triple-negative breast cancer

Abstract Background: Tumor-infiltrating lymphocytes (TILs) are a useful prognostic factor and predictive biomarker of neoadjuvant chemotherapy treatment response for breast cancer, especially in human epidermal growth factor receptor type 2 (HER2)-positive breast cancer and triple-negative breast cancer (TNBC). However, due to heterogeneity of TIL expression and distribution in the tissues, accurately predicting TIL expression, especially using limited core-needle biopsy specimens, is difficult. Therefore, an accurate and simple preoperative evaluation method is needed. We have reported that the TILs-ultrasonography (US) score determined by characteristic US findings has a predictive performance for lymphocyte-predominant breast cancer (LPBC). This study aimed to investigate whether the TILs-US score with added vascularity assessment has a better predictive performance for LPBC. Methods: This multicenter, retrospective study investigated the validation and scoring of the LPBC characteristic imaging findings and applied it for LPBC and non-LPBC prediction. A total of 100 patients with HER2-positive breast cancer (n = 59) and TNBC (n = 41) treated by curative surgery between January 2014 and December 2021 were evaluated. Stromal lymphocytes in surgical pathological specimens were evaluated; the cutoff value for predicting LPBC was defined as ≥50% stromal TILs. Preoperative US was examined for TIL indicators. The US images with characteristic TILs were scored for LPBC prediction. Univariate and multivariate logistic regression analyses were employed for each potential predictor variable of LPBC. Results: A total of 40 patients with ≥50% stromal TILs were defined as having LPBC. The examined characteristic US findings for predicting LPBC, shape (more lobulated), internal echo level (weaker), posterior echoes (stronger), and vascularity assessment (hypervascularity), were significantly associated with LPBC and used to assign the scoring for predicting LPBC. As previously reported, the TILs-US score ranged from 0–7 points based on three ultrasonic tissue characteristics: shape (round, oval, and polygonal or irregular, 0 points; lobulated, 1 point; and small lobulated, 2 points), internal echo level (high or equal, 0 points; low, 1 point; and extremely low, 2 points), and posterior echoes (shadowing or attenuating, 0 points; no change, 1 point; accentuated, 2 points; and extremely accentuated, 3 points). We further added vascularity assessment (avascular or hypovascular, 0 point; moderately vascular, 1 point; and hypervascular, 2 points) to this scoring system. Based on the receiver operating characteristics (ROC) curves (AUC [Area Under the Curve] 0.77), the score cutoff for predicting LPBC was 4 points for TILs-US score (sensitivity, 0.83; specificity, 0.55; and accuracy, 0.66). Multivariate logistic regression analysis revealed that cT (&amp;lt; T2), estrogen receptor (ER) negativity, and a TILs-US score of ≥4 points were significant LPBC predictors (odds ratio [OR] 3.60; p = 0.028; OR 8.68; p = 0.020; OR 5.99; p = 0.005). Conversely, based on the ROC curves (AUC 0.78), the score cutoff for predicting LPBC was 5 points after adding vascularity assessment (sensitivity, 0.93; specificity, 0.57; and accuracy, 0.71). Multivariate logistic regression analysis revealed that cT (&amp;lt; T2), ER negativity, and a TILs-US score adding vascularity assessment of ≥5 points were significant LPBC predictors (OR 5.12; p = 0.010; OR 10.3; p = 0.019; OR 20.1; p &amp;lt; 0.001). Conclusions: Including the vascularity assessment to the TILs-US score, which can be noninvasively obtained using US, is a more accurate preoperative predictor of LPBC. Vascularity assessment may be an auxiliary factor in predicting LPBC. Univariate and multivariate logistic analysis of significant clinicopathological factors predicting lymphocyte-predominant breast cancer. Citation Format: Yuri Kimura, Norio Masumoto, Sadako Akashi-Tanaka, Kayo Fukui, Midori Noma, Aya Nagata, Takashi Nakamura, Hiroaki Shima, Toshitaka Okuno, Akari Murakami, Yoshiaki Kamei, Shogo Nakano, Koji Arihiro. The TILs-US score adding vascularity assessment based on ultrasonography for predicting tumor-infiltrating lymphocytes in human epidermal growth factor receptor 2-positive and triple-negative breast cancer [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 P3-04-10.

Abstract P3-05-18: Unravelling the clinicopathological and functional significance of Replication Protein A (RPA) heterotrimeric complex in sporadic breast cancers

Abstract Unravelling the clinicopathological and functional significance of Replication Protein A (RPA) heterotrimeric complex in sporadic breast cancers Mashael Algethami1, Michael S Toss 1,4, Juliette Brownlie 1, Corinne L Woodcock 1,2, Chandar Jaipal1, Ahmed Shoqafi 1, Katia A Mesquita1, Adel Alblihy1,3, Andrew R Green1, Nigel P Mongan 1,5, Emad A Rakha 1,4, Jennie N Jeyapalan 1,2 and Srinivasan Madhusudan1,6 1 Nottingham Biodiscovery Institute, Academic Unit for Translational Medical Sciences, School of Medicine, University of Nottingham, University Park, Nottingham NG7 2RD, UK. 2 Faculty of Medicine and Health Sciences, Centre for Cancer Sciences, University of Nottingham, Sutton Bonington Campus, Sutton Bonington, Leicestershire LE12 5RD, UK 3 Medical Center, King Fahad Security College (KFSC), Riyadh 11461, Saudi Arabia. 4 Department of Pathology, Nottingham University Hospital, City Campus, Hucknall Road, Nottingham NG5 1PB, UK. 5 Department of Pharmacology, Weill Cornell Medicine, New York, NY, 10065, USA 6 Department of Oncology, Nottingham University Hospitals, Nottingham NG5 1PB, UK. Introduction: Replication Protein A (RPA) is a critical single-stranded DNA (ssDNA)-binding protein that coats and protects exposed ssDNA from endogenous nucleases. RPA is a heterotrimeric complex consisting of RPA1 (70kDa), RPA2 (32kDa), and RPA3 (14kDa) subunits. RPA provides a platform for recruitment of factors required during replication, checkpoint regulation, DNA repair [including homologous recombination (HR), nucleotide excision repair (NER), base excision repair (BER) and mismatch repair (MMR)], telomere maintenance and retro-transposition. To understand the role of RPA in breast cancer pathogenesis we conducted a comprehensive genomic, transcriptomic, bioinformatic, proteomic and preclinical study. Patients and methods: RPA1, RPA2 and RPA3 protein expression was evaluated in 4221 primary invasive breast carcinomas and 776 breast ductal carcinoma in situ (DCIS). Transcriptomic investigation was completed in the METABRIC cohort (n=1980). Detailed bioinformatics was conducted in the TCGA cohort (n=1090). RPA deficient breast cancer cell lines were tested for Cisplatin, Palbociclib and Olaparib sensitivity. Results: RPA1, RPA2 and RPA3 loss is frequent in DCIS and linked to aggressive phenotype (including high grade, ER and PR negativity) (all p values &amp;lt; 0.01). DCIS with low RPA1 was also associated with poor local recurrence-free interval (p&amp;lt; 0.00001). In invasive breast cancers, low RPA1, low RPA2 and low RPA3 were all associated with larger size, lympho-vascular invasion, higher histological grade, high stage, ER negativity and poor breast cancer specific survival. Transcriptomic alterations in low RPA tumors included those genes involved in steroid hormone biosynthesis, chemical carcinogenesis, and drug metabolism. Pre-clinically, RPA deficient breast cancer cells were sensitive to Cisplatin and Palbociclib (CDK4/6 inhibitor) therapy compared to controls. Additionally, the PARP1 inhibitor Olaparib was synthetically lethal in RPA1 and RPA2 deficient cells compared to controls. Increased Olaparib sensitivity was associated with double strand breaks, S-phase cell cycle arrest and increased apoptosis. Conclusions: We provide the first comprehensive evidence that RPA loss is an early event during breast cancer pathogenesis and promotes aggressive phenotypes. Pre-clinically RPA deficient breast cancer cells were selectively toxic to Cisplatin, Palbociclib and Olaparib. Citation Format: Mashael A. Algethami, Michael Toss, Juliette Brownlie, Corinne L. Woodcock, Chandar Jaipal, AHMED SHOQAFI, Katia Mesquita, Adel Alblihy, Nigel Mongan, Emad Rakha, Jennie N. Jeyapalan, Srinivasan Madhusudan, Andrew R. Green. Unravelling the clinicopathological and functional significance of Replication Protein A (RPA) heterotrimeric complex in sporadic breast cancers [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 P3-05-18.

Abstract P3-03-28: Clinicopathological Risk Factors of Brain Metastasis in Breast Cancer Patients: A Systematic Review and Meta-analysis

Abstract BACKGROUND: Breast cancer brain metastasis (BCBM) has an incidence rate of 5.1% among Breast Cancer (BC) patients. In BCBM, cancerous cells come from a primary tumor that implant and grow in the brain leading to potentially lethal neurologic symptoms and signs. With more clinical options and therapeutics strategies becoming available, a multi-disciplinary approach for treatment is needed in order to best meet BCBM patients’ needs, however, systematic guidelines for the screening of high-risk asymptomatic patients are still lacking and the BCBM diagnoses are performed only after symptoms manifestation. Therefore, understanding the clinical and pathologic drivers of BCBM aids in improving medical interventions by guiding future research directions. This study aims to determine the clinicopathological risk factors of brain metastasis (BM) among BC patients. METHODS: Retrospective cohort studies on the clinicopathological characteristics of BM versus non-BM (NBM) patients were retrieved through reputable search databases. Review Manager version 5.4.1 was used for statistical analyses. In the presence of heterogeneities, a pragmatic approach was undertaken to employ both random-effects (RE) and fixed-effect (FE) meta-analyses. The statistical significance of the pooled effect estimates was examined by the Z-test. Interstudy variations and heterogeneities were estimated using Cochran’s Q-statistic with P &amp;lt; 0.05 indicating a statistically significant heterogeneity. The risk of bias and the quality of studies were assessed at a study level using ROBINS-I tool. RESULTS: A total of four (4) retrospective cohort studies were included in the analyses. Random-effects meta-analyses with i2 &amp;lt; 50% (P &amp;gt; 0.05) had shown significant association (P &amp;lt; 0.05) on the increased incidences of BCBM development for HER2+ expression [OR: 2.43, 95% CI: 1.88 – 3.12, P &amp;lt; 0.00001] and for pre-menopausal status [OR: 1.77, 95% CI: 1.13 – 2.76, P = 0.01]. Fixed-effect meta-analyses had shown significant association (P &amp;lt; 0.05) on the decreased incidence of BCBM development for TNBC disease [OR: 0.66; 95% CI: 0.47 – 0.92; P = 0.02]. The risk of bias was low to moderate in the majority of studies. CONCLUSION: HER2 positivity and pre-menopausal status are risk factors for increased BCBM development while the absence of regional lymph node metastases and ILC findings are less likely to progress to BCBM. Higher T and N categories, higher histological grade, ER negativity, and PR negativity may be associated with higher risks of BCBM while lower T categories, HER2 negativity, and TNBC may likely be associated to less BCBM development. More powerful relevant and upcoming randomized clinical trials with larger sample sizes among BCBM patients versus non-BM patients must be made exploring certainty on the clinicopathological factors contributing to BM among BC patients. Keyword/s: risk factors, breast cancer, brain metastasis Citation Format: Korina Blanca C. Garcia, April Ann A. Lubay. Clinicopathological Risk Factors of Brain Metastasis in Breast Cancer Patients: A Systematic Review and Meta-analysis [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 P3-03-28.