Disease-free Survival In Breast Cancer Patients Research Articles

Abstract B038: CD4Th1 cytokine interferon gamma regulates genome profiles and inhibits tumorigenesis of disseminated cancer cells in breast cancer

Abstract Detection and persistence of disseminated cancer cells (DCCs) in the bone marrow (BM) of breast cancer (BC) patients has been identified as a primary source for late recurrence and distant metastasis in multiple organs and the central nervous system. Selective targeting and eradication of DCCs in BM can be of a significant advantage to prevent future recurrence and enhance disease-free survival in BC patients. Unfortunately, no such targeted therapy presently exists. Instead, intense chemotherapy in the adjuvant or neoadjuvant setting are given with the ultimate goal to prevent DCC-driven metastasis in these patients. Clinical studies have confirmed that DCCs have not been effectively eradicated by conventional chemotherapies. DCCs express unique gene signatures that differ from circulating tumor cells (CTC) in BC, but only limited knowledge is available for the genome profiles of DCCs compared to primary and metastatic lesions in BC patients. Here we investigated the detailed gene expression profiles of DCCs compared to primary and metastatic lesions from a murine spontaneous metastasis model and BC patients. Using a two-step immunomagnetic enrichment assay, we were able to successfully isolate highly enriched EpCAM+ DCCs from both the BM aspirates of HER2+ early BC patients and BALB-HER2/neu transgenic (BALB-neuT) spontaneous metastasis mice. RNA sequencing revealed that DCCs from HER2+ BC patients and BALB-neuT mice displayed remarkably similar genomic expression profiles, but both had substantially different profiles when comparing to primary and metastatic lesions. We identified highly enriched signatures in DCCs associated with cancer stemness, epithelial to mesenchymal transition (EMT), cell cycle, cell adhesion, progesterone signaling and cholesterol biosynthesis. Notably, these DCCs showed loss of MHC class I, MHC class II, and CD1d expression leading to a defect in antigen presentation machinery and their subsequent escape from CD4 T, CD8 T, B and NKT-cell mediated clearance. Interestingly, we observed for the first time that anti-tumor CD4Th1 cells, through interferon gamma (IFN-γ) production, upregulated CD1d, MHC class I and II expressions in DCCs leading to high infiltration of CD4Th1, NKT and B cells into the BM to selectively target and eliminate DCC burden. IFN-γ remarkably downregulated stemness, EMT, self-renewing potential and their genome signatures and induced apoptosis and irreversible senescence in DCCs of HER2+ BC patients. Notably, these IFN-γ treated patient DCCs lost their ability to grow into tumors in an immunodeficient NSG mouse model. Collectively, these results provide a detailed genome profile of DCCs from HER2+ BC patients and a novel role for anti-tumor CD4Th1 response in inhibiting DCC-driven tumorigenesis and metastasis. Citation Format: Ganesan Ramamoorthi, Marie Catherine Lee, Krithika Kodumudi, Colin Snyder, Namrata Gautam, Saurabh K. Garg, Shari Pilon-Thomas, Brian Czerniecki. CD4Th1 cytokine interferon gamma regulates genome profiles and inhibits tumorigenesis of disseminated cancer cells in breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr B038.

Prognostic role of the systemic immune-inflammation index and pan-immune inflammation value for outcomes of breast cancer: a systematic review and meta-analysis.

This review examined the literature for evidence on the prognostic ability of systemic immune-inflammation index (SII) and pan-immune inflammation value (PIV) for predicting overall survival (OS) and disease-free survival (DFS) in breast cancer patients. PubMed, Embase, Scopus, and Web of Science were searched with Google Scholar for gray literature. All types of studies reporting the association between SII or PIV and OS or DFS of breast cancer were eligible. 13 studies on SII and 4 studies on PIV were included. Meta-analysis showed that a high SII was a significant predictor of OS (HR: 1.97 95% CI: 1.54, 2.52 I2=76%) and DFS (HR: 2.07 95% CI: 1.50, 2.86 I2=79%) in breast cancer patients. These results did not change on sensitivity analysis and were more or less stable on multiple subgroup analyses. Pooled analysis showed that high PIV was also a significant predictor of poor OS (HR: 2.63 95% CI: 1.46, 4.74 I2=71%) and DFS (HR: 1.64 95% CI: 1.23, 2.17 I2=0%) in breast cancer patients. High SII and PIV can predict poor OS and DFS in breast cancer patients. High heterogeneity and the observational nature of data are important limitations of the review. Further studies are needed specifically on PIV to increase the strength of the evidence.

Unraveling the Angiogenic Puzzle: Pre-Treatment sVEGFR1 and sVEGFR2 Levels as Promising Prognostic Indicators in Early-Stage Breast Cancer Patients.

Despite the advancements in breast cancer (BrC) diagnosis and treatment, a considerable proportion of patients with early-stage disease still experience local recurrence or metastasis. This study aimed to assess the levels of specific angiogenic parameters in the EDTA plasma of BrC patients before and after treatment and to explore their clinical and prognostic significance. The levels of vascular endothelial growth factor A (VEGF-A), soluble form of vascular endothelial growth factor receptor type 1 (sVEGFR1), and soluble form of vascular endothelial growth factor receptor type 2 (sVEGFR2) were measured in 84 early BrC patients, both prior to surgery and within a median time of nine months post-treatment. Prognostic significance was evaluated using Kaplan-Meier survival and Cox regression analyses. Linear regression models were employed to examine the independent impact of selected angiogenic factors on DFS in breast cancer patients. The results of uni- and multivariate analyses indicated that a pre-treatment concentration of sVEGFR1 above 30.99 pg/mL was associated with improved disease-free survival (DFS) (p < 0.0001 for both analyses), while a pre-treatment concentration of sVEGFR2 above 9475.67 pg/mL was associated with an increased risk of BrC relapse (p < 0.0001 for both analyses). Additionally, a post-treatment concentration of sVEGFR2 above 7361.71 pg/mL was associated with better overall survival (OS) based on the Kaplan-Meier survival analysis (p = 0.0141). Furthermore, linear regression models revealed a significant inverse association between pre-treatment levels of sVEGFR1 and the risk of relapse (standardized β -0.2578, p = 0.0499) and a significant positive association of VEGF-A levels with the risk of recurrence (standardized β 0.2958, p = 0.0308). In conclusion, the findings suggest that both pre- and post-treatment levels of sVEGFR1 and sVEGFR2 may hold promise as potential prognostic markers for BrC patients.

Low pretreatment prognostic nutritional index predicts poor survival in breast cancer patients: A meta-analysis.

Prognostic nutritional index (PNI), as an indicator of nutritional immune status, has been shown to be associated with therapeutic effects and survival of solid tumors. However, the prognostic role of PNI before treatment in human breast cancer (BC) is still not conclusive. Hence, we performed this meta-analysis to assess the value of it in prognosis prediction for BC patients. We searched PubMed, Embase, Web of Science and EBSCO to identify the studies evaluating the association between PNI and survival such as overall survival (OS), disease-free survival (DFS) of BC, and computed extracted data into hazard ratios (HRs) for OS, DFS and clinicopathological features with STATA 12.0. A total of 2322 patients with BC from 8 published studies were incorporated into this meta-analysis. We discovered that low pretreatment PNI was significantly associated with worse OS, but not with DFS in BC patients. In stratified analyses, the result showed that decreased PNI before treatment was remarkably related with lower 3-year, 5-year, 8-year and 10-year OS, but not with 1-year survival rate in BC. In addition, although reduced PNI could not impact 1-year, 3-year or 5-year DFS, it considerably deteriorated 8-year and 10-year DFS in patients. Low pretreatment PNI deteriorated OS, 8-year and 10-year DFS in BC patients, implicating that it is a valuable prognostic index and improving the nutritional immune status may offer a therapeutic strategy for these patients.

Prognostic significance of prognostic nutritional index and systemic immune‐inflammation index in patients after curative breast cancer resection: a retrospective cohort study

BackgroundNutritional status and inflammation are closely associated with poor outcome in malignant tumors. However, the prognostic impact of postoperative in these variables on breast cancer (BC) remains inconclusive. We aimed to determine whether prognostic nutritional index (PNI), systemic immune‐inflammation index (SII), neutrophil–lymphocyte ratio (NLR) and platelet–lymphocyte ratio (PLR) affect two long-term outcomes among patients after curative resection of BC.MethodsWe retrospectively reviewed 508 patients with BC treated with curative surgery between February 5, 2013 and May 26, 2020. All patients were divided into 3 groups based on tertiles (T1-T3) of PNI, SII, NLR, and PLR. The effects of four indexes on disease-free survival (DFS) and overall survival (OS) have been evaluated using Cox proportional hazards models and Kaplan–Meier method.ResultsCompared with PNI-lowest cases, patients with highest PNI showed significantly longer DFS (multivariate adjusted hazard ratio [HR] = 0.37, 95% confident interval [CI] 0.19–0.70, P for trend = 0.002), whereas higher PLR seemed to be marginally associated with poorer DFS (P for trend = 0.086 and 0.074, respectively). Subgroup analyses indicate the potential modification effects of family history of BC and radiotherapy on the prognosis value of PNI to DFS in BC patients (P for interaction = 0.004 and 0.025, respectively). In addition, the levels of three inflammatory indices, namely SII, NLR, and PLR might be positively related with increased age at diagnosis (all P for trend < 0.001).ConclusionsA high PNI was associated with better DFS, supporting its roles as prognostic parameters for patients with BC. The nutritional status and systemic immune may exert great effects on patient prognosis. Further studies are warrant to explore the prognosis value of PLR.

Overexpression of PBK/TOPK relates to poor prognosis of patients with breast cancer: a retrospective analysis

BackgroundPDZ-binding kinase/T-lymphokine-activated killer cell-derived protein kinase (PBK/TOPK) is a potential prognostic indicator for patients with breast cancer. The objective of the present study was to explore the relationship between PBK/TOPK expression and clinicopathological indicators as well as the survival of patients with breast cancer.MethodsImmunohistochemical staining was used to detect the expression of PBK/TOPK in 202 cases of breast cancer tissues. The relationship between PBK/TOPK and clinicopathological parameters was evaluated using Spearman’s rank-order correlation. The difference in PBK/TOPK expression among different molecular types was analyzed with the chi-square test. Kaplan-Meier analysis was used to create a survival curve and the log rank test was used to analyze the overall survival (OS) and disease-free survival (DFS). Prognostic correlation was assessed using univariate and multivariate Cox regression analyses.ResultsAmong 202 breast cancer samples, PBK/TOPK was expressed (“+” and “++”) in 182 samples (90.1%). In addition, the histological grade, TNM stages, lymph node metastasis, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER-2), and Ki-67 were positively associated with PBK/TOPK expression. With regard to the molecular type, the expression of PBK/TOPK is different. The expression level of PBK/TOPK was negatively correlated with both the OS and DFS of breast cancer patients. The difference in the above results is meaningful (P < 0.05).ConclusionsPBK/TOPK is overexpressed in breast cancer, and the expression is closely related to the clinicopathological characteristics of the disease. Breast cancer patients with high expression of PBK/TOPK have a poor prognosis. Therefore, healthcare providers can optimize breast cancer management using this indicator.

O-195 Disease-free survival is not impacted by fertility preservation techniques for breast cancer women

Abstract Study question Do fertility preservation (FP) strategies using ovarian stimulation or not, impact long-term disease-free survival of breast cancer (BC) patients? Summary answer The disease-free survival of BC patients is not impacted by FP techniques whatever the timing of chemotherapy (neoadjuvant /adjuvant) and the use of ovarian stimulation. What is known already Fertility is often impaired in young women treated for BC. Therefore, FP has become a major issue in this population. Cryopreservation of oocytes or embryos after controlled ovarian hyperstimulation (COH) represents the most established method in this clinical situation. However, the hormonal consequences of COH protocols still raise safety concerns, often leading oncologists to contraindicate the use of this FP technique. Although alternative FP options without exogenous hormone administration may be considered, they remain suboptimal for treating the putative future infertility. Study design, size, duration Retrospective bicentric cohort study including including 740 BC women, 18-43 years of age, referred for FP between 2013 and 2019. Participants/materials, setting, methods Overall, 328 underwent at least one ovarian stimulation cycle (STIM group) and 412 had a technique without hormonal administration (No STIM group). Log Rank test was used to compare both groups and Cox proportional-hazard model was applied for multivariable analyses. Main results and the role of chance Women of the No STIM group were significantly younger and present with more severe disease. Follow-up data for recurrences were available for 80.9% of the cohort and the median time to follow-up was 4.2 [2.9-5.8] vs. 5.6 [4.1-6.7] years between STIM and No STIM group (p &amp;lt; 0.0001). According to log-rank test, the risk of recurrence did not differ between the two groups (p = 0.09) even after adjustment on age, SBR grade, triple negative status or type of planned chemotherapy. Limitations, reasons for caution The retrospective nature of the study represents the main limitation. The median follow-up of 4.2 to 5.6 years, might be considered short to assess the long-term safety of FP techniques. The heterogeneity between oncological centers regarding the use of COH and specific protocols may lalso be a reason for caution. Wider implications of the findings Our findings provide reassuring safety data on the use COH for FP in BC patients, whatever the timing of chemotherapy. However, further investigations with a longer follow-up are needed to definitely consider COH safe in particular in neoadjuvant situations. In addition, the benefit of letrozole supplementation during COH requires confirmation. Trial registration number Not applicable

Metformin counteracts stimulatory effects induced by insulin in primary breast cancer cells

BackgroundMetabolic disorders are associated with increased incidence, aggressive phenotype and poor outcome of breast cancer (BC) patients. For instance, hyperinsulinemia is an independent risk factor for BC and the insulin/insulin receptor (IR) axis is involved in BC growth and metastasis. Of note, the anti-diabetic metformin may be considered in comprehensive therapeutic approaches in BC on the basis of its antiproliferative effects obtained in diverse pre-clinical and clinical studies.MethodsBioinformatics analysis were performed using the information provided by The Invasive Breast Cancer Cohort of The Cancer Genome Atlas (TCGA) project. The naturally immortalized BC cell line, named BCAHC-1, as well as cancer-associated fibroblasts (CAFs) derived from BC patients were used as model systems. In order to identify further mechanisms that characterize the anticancer action of metformin in BC, we performed gene expression and promoter studies as well as western blotting experiments. Moreover, cell cycle analysis, colony and spheroid formation, actin cytoskeleton reorganization, cell migration and matrigel drops evasion assays were carried out to provide novel insights on the anticancer properties of metformin.ResultsWe first assessed that elevated expression and activation of IR correlate with a worse prognostic outcome in estrogen receptor (ER)-positive BC. Thereafter, we established that metformin inhibits the insulin/IR-mediated activation of transduction pathways, gene changes and proliferative responses in BCAHC-1 cells. Then, we found that metformin interferes with the insulin-induced expression of the metastatic gene CXC chemokine receptor 4 (CXCR4), which we found to be associated with poor disease-free survival in BC patients exhibiting high levels of IR. Next, we ascertained that metformin prevents a motile phenotype of BCAHC-1 cells triggered by the paracrine liaison between tumor cells and CAFs upon insulin activated CXCL12/CXCR4 axis.ConclusionsOur findings provide novel mechanistic insights regarding the anti-proliferative and anti-migratory effects of metformin in both BC cells and important components of the tumor microenvironment like CAFs. Further investigations are warranted to corroborate the anticancer action of metformin on the tumor mass toward the assessment of more comprehensive strategies halting BC progression, in particular in patients exhibiting metabolic disorders and altered insulin/IR functions.

Breast Cancer Classification Based on Tumor Budding and Stem Cell-Related Signatures Facilitate Prognosis Evaluation.

BackgroundTumor budding (TB) is emerging as a prognostic factor in multiple cancers. Likewise, the stemness of cancer cells also plays a vital role in cancer progression. However, nearly no research has focused on the interaction of TB and tumor stemness in cancer.MethodsTissue microarrays including 229 cases of invasive breast cancer (BC) were established and subjected to pan-cytokeratin immunohistochemical staining to evaluate molecular expression. Univariate and multivariate analyses were applied to identify prognostic factors of BC, and the Chi-square test was used for comparison of categorical variables.ResultsHigh-grade TB was significantly associated with T stage, lymph node metastasis, tumor node metastasis (TNM) stage, epithelial-mesenchymal transition, and poor disease-free survival (DFS) of BC patients. We also found that the prognostic value of TB varied widely among different subtypes and subgroups. Cox regression analysis then showed that TB grade was an independent prognostic factor. Moreover, cancer stem cell (CSC) markers CD44 and ALDH1A1 were significantly higher in high-grade TB tumors. Consequently, patients were classified into high CSC score subgroup and low CSC score subgroups. Further research found that CSC scores correlated with clinicopathological features and DFS of BC patients. Based on TB grade and CSC scores, we classified BC patients into TBlow-CSCslow (type I), TBlow-CSCshigh (type II), TBhigh-CSCslow (type III), and TBhigh-CSCshigh (type IV) subgroups. Survival analysis showed that patients in the type I subgroup had the best DFS, whereas those in the type IV subgroup had the worst DFS. Finally, a TB-CSC-based nomogram for use in BC was established. The nomogram was well calibrated to predict the probability of 5-year DFS, and the C-index was 0.837. Finally, the area under the curve value for the nomogram (0.892) was higher than that of the TNM staging system (0.713).ConclusionThe combination of TB grade with CSC score improves the prognostic evaluation of BC patients. A novel nomogram containing TB grade and CSC score provides doctors with a candidate tool to guide the individualized treatment of cancer patients.

Impact of Androgen Receptor Expression and the AR:ER Ratio on the Survival Outcomes in the Diverse Subgroups of Vietnamese Breast Cancer: A Single Institutional Retrospective Cohort Analysis.

Background: The androgen receptor (AR) has recently emerged as a useful marker for the more favorable prognosis and better outcomes among women with estrogen receptor (ER) + ve breast cancer (BC) and the further refinement of BC subtype. Furthermore, AR expression in ER − ve tumors has a particular prognostic significance. Additionally, the ratio of nuclear AR to ER may critically have an influence on tumor biology and respond to endocrine therapy. Purpose: To define the AR expression and AR:ER ratio, and explored their correlation with the clinicopathological features, prognosis, and survival outcomes in the various subclasses of invasive BC. Methods: The current study was conducted on 522 BC patients who had surgical operations, without neoadjuvant chemotherapy by applying a retrospective cohort analysis. The clinicopathological characteristics were recorded. Immunohistochemical staining was performed on AR, ER, PR, HER2, and Ki67. Expression of AR was paired into different immunophenotypes for analysis with clinicopathological features and survival. All BC patients’ survival was analyzed using Kaplan–Meier and log-rank models. Results: The presence of AR was detected in 65.3%. Positive AR, the ratio of AR:ER<2, luminal androgen receptor (LAR) + and AR + HER2 + immunophenotypes were significantly associated with better prognostic features. AR:ER<2 was observed in the prolonged overall survival (OS) and disease-free survival (DFS) (87.9 and 86.2%, respectively) compared to AR:ER≥2 (25.0% in both) (P < .001). In contrast, in HR + ve BCs, the AR expression was not significantly correlated with survival. The multivariate model revealed that the ratio of nuclear AR to ER remained as an independent prognostic variable. Conclusion: The AR expression had a distinct OS and DFS. The AR:ER ratio is an independent indicator for predicting the OS and DFS of BC patients in both univariate and multivariate analyses.

Prognostic Value of Pretreatment Lymphocyte-to-Monocyte Ratio and Development of a Nomogram in Breast Cancer Patients.

PurposeThe objective of this study was to explore the prognostic significance of pretreatment hematologic parameters in predicting disease-free survival (DFS) of breast cancer patients.Materials and MethodsThe medical records of 440 breast cancer patients in Shandong Cancer Hospital and Institute from 2003 to 2013 were analyzed retrospectively. Through the results of blood routine before treatment, the absolute lymphocyte count (ALC), absolute neutrophil count (ANC), absolute monocyte count (AMC), and absolute platelet count (APC) in peripheral blood were collected. The lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and neutrophil-to-monocyte ratio (NMR) were calculated. Cox proportional hazard model was used for univariate and multivariate analysis. The DFS was compared using Kaplan–Meier method. The prognostic nomogram of patients with breast cancer was developed.ResultsThe median DFS for all patients was 64.10 months. Univariate analysis showed that the DFS was associated with surgical approach, TNM stage, molecular subtype, neoadjuvant chemotherapy, radiotherapy, and LMR (p < 0.05). TNM stage, molecular subtype, and LMR were independent prognostic factors of breast cancer in multivariate analysis (p < 0.05). According to the Kaplan–Meier survival curve analysis, patients with higher LMR (≥4.85) were associated with longer median DFS (median DFS, 85.83 vs. 60.90, p < 0.001). The proposed nomogram that incorporated LMR, TNM stage, and molecular subtype got a concordance index (c-index) of 0.69 in predicting 5-year DFS.ConclusionIn breast cancer patients, higher LMR was associated with longer median DFS and the nomogram including LMR, TNM stage, and molecular subtype could accurately predict the prolonged 5-year DFS of breast cancer patients.

Identification of a Transcription Factor Signature That Can Predict Breast Cancer Survival.

Background The expression pattern of transcription factors (TFs) can be used to develop potential prognostic biomarkers for cancer. In this study, we aimed to identify and validate a TF signature for predicting disease-free survival (DFS) of breast cancer (BRCA) patients. Methods Lasso and the Cox regression analyses were applied to construct a TF signature based on a gene expression dataset from TCGA. The prognosis value of the TF signature was investigated in the TCGA database, and its reliability was further validated in 3 independent datasets from Gene Expression Omnibus (GEO). The prognosis performance of the TF signature was compared with 4 previously published gene signatures. To investigate the association between the TF signature and hallmarks of cancer, Gene Set Enrichment Analysis (GSEA) was carried out. The correlations of the TF signature and the levels of immune infiltration were also investigated. Results An 11-TF prognostic signature was constructed with good survival prediction performance for BRCA patients. By using the risk score model based on the 11-TF signature, BRCA patients were stratified into low- and high-risk groups and showed good and poor disease-free survival (DFS), respectively. The risk score was an independent prediction indicator when adjusting for other clinicopathological factors. Furthermore, the 11-TF signature had a better survival prediction performance compared to 4 previously published gene signatures. Moreover, the risk score was a cancer hallmark. Finally, a high-risk score was associated with higher infiltration of M0 and M2 macrophages and was associated with a lower infiltration of resting memory CD4+ T cells and CD8+ T cells. Conclusion The findings in this study identified and validated a novel prognostic TF signature, which is an independent biomarker for the prediction of DFS in BRCA patients.

Abstract PS6-59: Impact of Cholecystectomy in Breast Cancer Recurrence

Abstract Background: Several published studies demonstrate that bile acid metabolites may influence the growth of breast cancer cells in vitro. Our group has shown reduced plasma concentrations of cholic acid and chenodeoxycholic acid (primary bile acids) and deoxycholic acid and lithocholic acid (secondary bile acids) in breast cancer patients that later on develop tumor recurrence. Cholecystectomy reduces the circulating bile acid pool. In patients with prior cholecystectomy, changes in bile acid metabolites may contribute to breast cancer tumorigenesis and recurrence. This study investigates our institutional rate of cholecystectomy in women diagnosed with breast cancer and its impact on breast cancer recurrence. Methods: A retrospective review of patients with an invasive breast cancer diagnosis between 2014-2015 was conducted. Demographics, preoperative variables, surgical history and clinical outcome data was collected. 5-year disease-free survival (DFS) was compared using a Log-rank (Mantel-Cox) test. Results: The study included 264 patients with mean age of 60.9. Most were Caucasian (83.5%). The majority were diagnosed at Stage II or lower (80.3%) and had hormone receptor positive, HER2 negative breast cancer (72.9%). Approximately 22.7% of patients had prior cholecystectomy surgery. The only statistically significant heterogeneity in demographic data between patients with and without cholecystectomy was body mass index (BMI). Patients with cholecystectomy had a mean BMI of 33.3, versus 29.1 in patients with intact gallbladders. The 5-year DFS in breast cancer patients with cholecystectomy was 91.6%, versus 97.1% in patients with intact gallbladders (p=0.06). Conclusion: Women with breast cancer who had a history of cholecystectomy had increased rates of breast cancer recurrence over a 5-year period compared to women with breast cancer with intact gallbladders. Although this result was not statistically significant, a trend was seen. Future study of a larger patient sample size sample may lead to a statistical significant difference. The statistically significant difference in BMI between the two patient groups is likely a confounding factor, given increased BMI is a known risk factor for developing cholecystitis and breast cancer. This data supports existing in vitro studies that bile acids may influence the growth of breast cancer cells. There may be utility in closer follow-up of women with breast cancer and a history of cholecystectomy given the increased rate of breast cancer recurrence in this population. Citation Format: Isheeta Madeka, Rohin Gawdi, Katherine L Cook, Akiko Chiba. Impact of Cholecystectomy in Breast Cancer Recurrence [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS6-59.

Identifying Biomarkers to Predict the Progression and Prognosis of Breast Cancer by Weighted Gene Co-expression Network Analysis.

Breast cancer (BC) is the leading cause of cancer death among women worldwide. The molecular mechanisms of its pathogenesis are still to be investigated. In our study, differentially expressed genes (DEGs) were screened between BC and normal tissues. Based on the DEGs, a weighted gene co-expression network analysis (WGCNA) was performed in 683 BC samples, and eight co-expressed gene modules were identified. In addition, by relating the eight co-expressed modules to clinical information, we found the blue module and pathological stage had a significant correlation (r = 0.24, p = 1e–10). Validated by multiple independent datasets, using one-way ANOVA, survival analysis and expression level revalidation, we finally screened 12 hub genes that can predict BC progression and prognosis. Functional annotation analysis indicated that the hub genes were enriched in cell division and cell cycle regulation. Importantly, higher expression of the 12 hub genes indicated poor overall survival, recurrence-free survival, and disease-free survival in BC patients. In addition, the expression of the 12 hub genes showed a significantly positive correlation with the expression of cell proliferation marker Ki-67 in BC. In summary, our study has identified 12 hub genes associated with the progression and prognosis of BC; these hub genes might lead to poor outcomes by regulating the cell division and cell cycle. These hub genes may serve as a biomarker and help to distinguish different pathological stages for BC patients.

Wntless (Wls): A Prognostic Index for Progression and Patient Survival of Breast Cancer.

BackgroundWntless (Wls) is an essential protein that is necessary for the secretion of Wnt proteins. While numerous researches have demonstrated that aberrations in Wnt/β-catenin expression lead to tumorigenesis and progression in many cancer types, the effects of Wls in breast cancer (BC) are less studied.MethodsThe mRNA and protein expression of Wls in BC cell lines were detected by RT-qPCR and Western blot; the protein expression of patient samples was detected by immunohistochemistry (IHC). The associations between Wls expression and clinicopathological factors as well as survival time, including overall survival (OS) and disease-free survival (DFS) were analyzed. Bioinformatics analysis was used to reveal the correlation between Wls genes and associated genes or pathways.ResultsWls was overexpressed in BC cell lines and tissues. The expression level of Wls was significantly correlated with tumor size, estrogen receptor (ER), progesterone receptor (PR), Ki-67, molecular classification, and follow-up status. Spearman correlation analysis showed that Wls protein expression was negatively correlated with ER and PR, which was confirmed by bioinformatics analysis in mRNA level. However, there were positive relationships with MBNG (modified Black’s nuclear grade), tumor size, Ki-67, molecular classification, follow-up, and vital status. Univariate and multivariate analysis showed that Wls was an independent prognostic factor for OS and DFS in BC patients. Moreover, Wls was a significant prognostic indicator of OS and DFS in a hormone receptor-positive (HR+) subgroup. GSEA showed that estrogen and androgen response, as well as epithelial-mesenchymal transition pathways, were up-regulated in the Wls high-expression group.ConclusionOverexpression of Wls is a significant marker of worse prognosis in BC and might play a crucial role in the HR+ subgroup.

Predictive and prognostic value of PDL1 protein expression in breast cancer patients in neoadjuvant setting

ABSTRACT Objective Programmed death-ligand-1 (PDL1) is a molecule involved in immune evasion in various kinds of tumors. Here, we aim to determine whether the expression of PDL1 protein is related to the response of patients to neoadjuvant therapy and survival outcome. Methods Immunohistochemistry (IHC) was performed on paraffin-embedded tumor samples from core needle biopsy before neoadjuvant therapy (NAT). Univariate and multivariate logistic regression were used to analyze the associations between PDL1 protein expression and pathological complete response (pCR) outcome. Kaplan-Meier plot and log-rank test were used to compare disease-free survival (DFS) between groups. A cox proportional hazards model was used to calculate the adjusted hazard ratio (HR) with 95% confidential interval (95%CI). Results A total of 94 patients were included for IHC testing. PDL1 protein expression on tumor cells was associated with better pCR rate in both univariate (OR = 2.621, p = 0.043) and multivariate (OR = 3.595, p = 0.029) logistic regression analysis. It was also associated with shorter DFS both by log-rank test (p = 0.015) and cox hazard model (HR = 22.824, 95%CI 1.621–321.284, p = 0.020). In hormone receptor (HR)-positive patients, PDL1 protein expression was also associated with better pCR (OR = 2.362, p = 0.022). It was also associated with poor DFS (HR = 18.821, 95%CI 1.645–215.330, p = 0.018). Conclusions Our results show that PDL1 protein expression is a predictive biomarker of pCR and a prognostic factor of DFS in breast cancer patients and HR-positive subgroups.

Importance of CD45RO+ tumor-infiltrating lymphocytes in post-operative survival of breast cancer patients.

In recent years, the prognostic/predictive significance of tumor infiltrating lymphocytes (TILs) has become a topic of interest. Here, we aimed to evaluate the prognostic significance of CD3+, CD8+, CD45RO+ and Foxp3+ TILs in breast cancer, as well as the relation of these markers to other clinicopathological features of this disease. FFPE tumor samples from 94 females with invasive ductal carcinoma of the breast were retrospectively selected and immunohistochemically assessed for CD3, CD8, CD45RO and Foxp3 expression. Digital photos were acquired from the center (CT) and invasive margins (IM) of the tumors, after which positive cells were counted using ImageJ software. We found that greater infiltrations of target lymphocyte subpopulations were associated with TNM stage III, lymph node metastasis, high histological grade, ER negativity and HER2 positivity. The ratios of CD8+ cytotoxic T cells to CD3+, CD45RO+ and Foxp3+ TILs were found to be relatively higher in tumors exhibiting the aforementioned characteristics. In univariate survival analyses, CD8+ TILs in the IM and total CD45RO+ TILs were found to be significantly associated with overall survival (OS). Infiltration of CD45RO+ TILs in the CT and lymph node status were variables that significantly correlated with disease-free survival (DFS). Multiple Cox regression analyses revealed independent significant prognostic effects of total CD45RO+ TILs and lymph node status (HR of 3.24 and 3.19, respectively) in predicting OS. Infiltration of CD45RO+ TILs in the CT (HR 3.12) and lymph node status (HR 3.15) also exhibited significant prognostic effects on DFS. From our data we conclude that CD45RO+ TILs serve as prognostic factors for predicting OS and DFS of breast cancer patients.

Impact of germinal center-associated nuclear protein polymorphisms on breast cancer risk and prognosis in a Japanese population.

Germinal center-associated nuclear protein (GANP) is a phosphoprotein involved in mRNA export and regulation of DNA recombination. Although GANP expression in human breast cancer tissue is associated with breast cancer prognosis, the association between the genetic background of GANP and susceptibility and prognosis of breast cancer is unclear. We selected 694 breast cancer cases and 1376 age- and menopausal status-matched non-cancer controls from the Hospitable-based Epidemiologic Research Program, conducted at Aichi Cancer Center between 2001 and 2005. We evaluated the impact of two polymorphisms at the GANP locus (rs2839178 and rs11702450) on the susceptibility and prognosis of breast cancer. Reference alleles were defined as the A allele for rs2839178 and G allele for rs11702450. The GG genotype of rs2839178 was statistically significantly associated with breast cancer risk (odds ratio [OR] 0.48, 95% confidence interval [CI] 0.30-0.76, P = 0.002). In prognostic analysis, compared to those with AA genotype of rs2839178, patients with AG or GG genotypes had longer disease-free survival (DFS) (hazard ratio [HR] 0.71, 95% CI 0.49-1.04 and HR 0.42, 95% CI 013-1.42, respectively, P for trend = 0.04). eQTL analysis indicated that association with rs2839178 can be explained by the effect of rs2839173 on expression of GANP/MCM3AP. The G allele of rs2839178 at the GANP locus was significantly associated with reduced breast cancer risk and longer DFS in breast cancer patients, showing a consistent direction in the association between susceptibility and clinical outcome. GANP is, therefore, important for the occurrence and progression of sporadic breast cancer.

Expression and prognostic value of HER-2/neu in primary breast cancer with sentinel lymph node metastasis.

The present study explores the correlation of human epidermal growth factor receptor-2 (HER-2) protein expression with sentinel lymph node (SLN) metastasis and prognosis of breast cancer. The breast cancer tissues and adjacent tissues were obtained from patients with primary breast cancer. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the mRNA level of HER-2. Spearman correlation analysis was used to analyze the correlation of HER-2 expression with SLN metastasis. The disease-free survival (DFS) and overall survival (OS) of breast cancer patients were investigated. Univariate and multivariate analyses were performed to explore factors influencing SLN metastasis and prognosis of breast cancer. Compared with adjacent tissues, HER-2 expression was significantly up-regulated in breast cancer tissues. HER-2 expression was correlated with the pathological type, tumor node metastasis (TNM) staging, histological grade, blood vessel invasion, SLN metastasis, estrogen receptor (ER), and progesterone receptor (PR). The expression level of HER-2 was positively related to the SLN metastasis (r=0.548). Median DFS and OS were longer in patients with negative HER-2 expression than in patients with positive HER-2 expression. TNM staging, SLN metastasis, and expression levels of HER-2 and ER were independent factors for DFS of breast cancer patients, while TNM staging, blood vessel invasion, histological grade, SLN metastasis, and expression levels of HER-2 and PR were independent factors for OS of breast cancer patients. Our study suggests that high expression of HER-2 promoted SLN metastasis. HER-2 expression and SLN metastasis were the independent factors for the prognosis of breast cancer.

Clinical features and prognosis of patients with first-episode liver metastasis of different molecular subtypes of breast cancer

To investigate the clinical features and prognosis of patients with first-episode liver metastasis of different molecular subtypes of breast cancer and risk factors for liver metastasis of breast cancer. A retrospective analysis was performed for 122 breast cancer patients with first-episode liver metastasis from January 2009 to January 2014. According to the cell surface receptors of breast cancer, these patients were divided into the four molecular subtypes of Luminal A, Luminal B, human epidermal growth factor receptor 2 (HER2) overexpression, and triple-negative breast cancer (TNBC). The association of patients' age at initial diagnosis, body mass index (BMI), menstruation status, clinical TNM (cTNM) stage, levels of lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) at recurrence, liver metastasis, and treatment condition with the patients' prognosis were analyzed. The chi-square test and Fisher's exact test were used for categorical data, the Kaplan-Meier method was used for survival analysis, the log-rank test was used for univariate analysis of influencing factors, and the Cox regression model was used for multivariate analysis. Among the 122 patients, 12 had Luminal A subtype, 61 had Luminal B subtype, 30 had HER2 overexpression subtype, and 19 had TNBC subtype. In the patients with Luminal A, Luminal B, HER2 overexpression, and TNBC subtypes, the median disease-free survival (DFS) was 32, 23, 16, and 10 months, respectively (P = 0.001), the median overall survival (OS) was 54, 35, 26, and 13 months, respectively (P = 0.003), and the median OS after liver metastasis was 30, 16, 10, and 9 months, respectively (P = 0.019). In HER2-positive patients, the application of trastuzumab in the past significantly prolonged the patients' DFS by 11 months and OS by 18 months (P < 0.05). The results of the multivariate analysis showed that cTNM stage, molecular subtype, and targeted therapy were independent influencing factors for DFS of breast cancer patients with liver metastasis (P < 0.05), and that BMI, increased LDH at recurrence, cTNM stage, molecular subtype, salvage chemotherapy, radiotherapy, and targeted therapy were independent influencing factors for OS of breast cancer patients with liver metastasis (P < 0.05). The patients with TNBC, HER2 overexpression, and Luminal B subtypes exhibited worse prognosis and had a risk of recurrence 15.97, 8.81, and 4.76 times higher than those with Luminal A subtype. The risk of death in the patients with TNBC, HER2 overexpression, and Luminal B subtypes was 8.42, 6.02, and 3.86 times that in those with Luminal A subtype. The prognosis of breast cancer patients with first-episode liver metastasis is associated with the increase in LDH when liver metastasis occurs, BMI, cTNM stage, and molecular subtype. Compared with the patients with Luminal subtypes, those with HER2 overexpression and TNBC subtypes tend to develop liver metastasis in early stage and have a shorter OS. Salvage chemotherapy, targeted therapy, and radiotherapy can significantly improve the prognosis of patients with liver metastasis.