Worse Breast Cancer Prognosis Research Articles

Abstract PO2-02-02: BREAst Cancer Personalized NuTrition (BREACPNT): dietary intervention in HR+ breast cancer patients treated with endocrine therapy

Abstract BACKGROUND Breast cancer patients treated with adjuvant endocrine therapy commonly experience weight gain, which has been associated with low adherence to therapy and worse breast cancer prognosis. Dietary interventions are the first-line treatment for weight management in breast cancer patients and have beneficial results. Yet, the ability to maintain these outcomes requires further research. Here, we aimed to assess whether a personalized postprandial glucose targeting (PPT) diet is beneficial for weight management and metabolic health as compared with the recommended Mediterranean (MED) diet in this population. DESIGN AND METHODS BREACPNT is a phase-2 randomized trial in HR+ patients with breast cancer, treated with adjuvant endocrine therapy. We randomly assigned participants to follow a MED diet or a PPT diet for a 6-month dietary intervention and additional 6-month follow-up. The PPT diet is based on a machine-learning algorithm that integrates clinical and microbiome features to predict personal postprandial glucose responses (PPGRs). During the intervention, participants were monthly followed by a registered dietitian, they were additionally connected to a 2-week continuous glucose monitoring (CGM) on 3-timepoints, provided stool and blood samples and self-reported dietary intake using a smartphone application. RESULTS Among 127 randomized patients (mean ± SD age 53 ± 10 years, BMI 28.6 ± 5 kg/m2), 103 (81%) completed the 6-month intervention period. A total of 94 participants provided 12-month follow-up data. Both interventions led to a significant weight loss. The PPT-diet showed numerically greater weight loss with median change of -1.45kg (IQR -3.75 – 0) compared to the MED-diet with median change of -1kg (IQR -3.2 - 0.1) , however this was not significant (p=0.9). Notably, at the 12-month time point , participants in the PPT arm were more likely to maintain the weight loss (median -1.2kg, IQR -4 - 0.1) compared to participants in the MED-arm (median -0.15kg , IQR -4.25 - 1.5), p=0.14) and had significantly improved waist circumference (median, IQR; -5 cm, -8-(-2) in the PPT arm, compared to -1cm, -6.5-3 in the MED arm, p=0.01). In terms of glucose metabolism the PPT-diet led to improved glycemic control as measured by CGM, with significant lower average postprandial glucose responses (PPGRs) and glucose coefficient of variation (CV) compared to the MED-diet arm (mean 6-month change in PPGR was 2.1 ± 7.5 mg/dL × h for MED and −3.4 ± 5.6 mg/dL × h for PPT, p< 0.001, and the 6-month mean change for CV was 0.15 ± 2.76 for MED and −1.37 ± 3.2 for PPT, p< 0.01). Within the entire cohort, participants who experienced the greatest weight loss (upper quartile) were more likely to be postmenopausal compared to participants in the lower quartile. Additionally, participants who did not achieve weight loss (lower quartile) were significantly more likely to be treated with tamoxifen than AI (p=0.015). CONCLUSIONS In this dietary intervention in HR+ breast cancer patients, both PPT diet and MED diet led to weight loss and improved food quality, with a slight advantage to PPT diet. PPT diet additionally improved glycemic features as compared to MED diet, which are linked to lower risk of obesity and disease recurrence, suggesting an advantage in a glucose targeting approach. Premenopausal and Tamoxifen treated patients were less likely to lose weight through the intervention. These findings may have implications for dietary advice in clinical practice. Citation Format: Michal Rein, Maya Dadiani, Anastasia Godneva, Michal Bakalenik-Gavry, Dana Morzaev-Sulzbach, Maya Lotan-Pompan, Adina Weinberger, Eran Segal, Einav Gal-Yam. BREAst Cancer Personalized NuTrition (BREACPNT): dietary intervention in HR+ breast cancer patients treated with endocrine therapy [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-02-02.

Expression and Clinical Significance of NUDCD1, PI3K/AKT/mTOR Signaling Pathway-Related Molecules and Immune Infiltration in Breast Cancer

BackgroundNUDCD1 (NudC Domain Containing 1) performs an essential function in biological processes such as cell progression, migration, cell cycle, and intracellular material transport. Many solid tumors express it highly, which is a prospective biomarker and therapeutic approach. However, the expression and clinical importance of NUDCD1 across breast cancer is unclear. MethodsThe expressions of NUDCD1 in breast cancers and normal breast tissues were studied utilizing the TIMER database and immunohistochemical analysis. Subsequently, we validate the association between the expression of NUDCD1 and clinicopathologic features and prognosis of breast cancer. The immunohistochemical experiments of pathway-related molecules were done on 214 breast cancer tissue microarrays. The investigation of correlation between NUDCD1 expression and tumor immune infiltration was subsequently conducted. ResultsThrough the utilization of bioinformatics analysis and immunohistochemical experiments, it was determined that NUDCD1 exhibited upregulation within breast cancer. Furthermore, it was discovered that an elevated expression of NUDCD1 may potentially be linked to a worse prognosis in breast cancer. Our study reveals that the PI3K/AKT/mTOR signaling pathway may perform a function in NUDCD1 regulating breast cancer progression via enrichment analysis. Furthermore, the expression of NUDCD1 may be associated with the degree of immunological infiltration. ConclusionThe expression of NUDCD1 was explored to be elevated in breast cancer and was observed to be correlated with a poorer prognosis. p-AKT, PI3K, AKT, mTOR, and p-mTOR expression levels underwent significant elevation in breast cancer. The function of NUDCD1 within breast cancer might be associated with the PI3K/AKT/mTOR signaling pathway.

Expression of semaphorin 3A (SEMA3A) in breast cancer subtypes

Breast cancer is a major health concern, and its accurate diagnosis and management depend on identifying its histological type and biological subtype. Semaphorin-3A (SEMA3A) is a membrane protein with diverse roles in cellular processes, including cancer progression and angiogenesis regulation. However, its role in breast cancer remains poorly understood. This study aimed to evaluate SEMA3A expression in breast cancer and investigate its distribution across breast cancer subtypes: luminal A, luminal B, HER2-positive, and triple-negative breast cancer (TNBC). Immunohistochemical evaluation was performed on 98 breast cancer patients' tumor specimens, and SEMA3A expression was assessed in tumor cells and vessels. The study included the analysis of the Ki67 proliferation index, estrogen receptor (ER) expression, progesterone receptor (PR) expression, and HER2 status in conjunction with SEMA3A expression. Analysis indicated positive expression of SEMA3A in breast cancer cells in 60 out of 98 cases. SEMA3A expression correlated positively with Ki67 levels in tumor cells (p = 0.0005, R Spearman 0.338). Notably, a negative correlation was found between SEMA3A expression and ER and PR levels in tumor cells (p = 0.04, Spearman's R = − 0.21 and p = 0.016, Spearman's R = − 0.25 respectively). HER2 status did not significantly influence SEMA3A expression. The study demonstrated positive SEMA3A expression in tumor vessels across all subtypes in 91 out of 98 cases, suggesting its involvement in endothelial cell function. However, no significant differences in SEMA3A expression were observed between breast cancer subtypes either in vessels or tumor cells. These findings suggest that elevated SEMA3A expression may be associated with worse prognosis in breast cancer, especially in ER- and PR-negative tumors. Further investigations are warranted to fully comprehend the role of SEMA3A in breast cancer biology, which may lead to the identification of novel therapeutic targets and personalized treatment strategies for breast cancer patients.

CDKN2A was a cuproptosis-related gene in regulating chemotherapy resistance by the MAGE-A family in breast cancer: based on artificial intelligence (AI)-constructed pan-cancer risk model.

Before the discovery of cuproptosis, copper-loaded nanoparticle is a wildly applied strategy for enhancing the tumor-cell-killing effect of chemotherapy. Although copper(ii)-related researches are wide, details of cuproptosis-related bioprocess in pan-cancer are not clear yet now, especially for prognosis and drug sensitivity prediction yet now. In this study, VOSviewer is used for the literature review, and R4.2.0 is used for data analysis. Public data are collected from TCGA and GEO, local breast cancer cohort is collected to verify the expression level of CDKN2A. 7036 published articles exhibited a time-dependent linear relationship (R=0.9781, p<0.0001), and breast cancer (33.4%) is the most researched topic. Cuproptosis-related-genes (CRGs)-based unsupervised clustering divides pan-cancer subgroups into four groups (CRG subgroup) with differences in prognosis and tumor immunity. 44 tumor-driver-genes (TDGs)-based prediction model of drug sensitivity and prognosis is constructed by artificial intelligence (AI). Based on TDGs and clinical features, a nomogram is (C- index: 0.7, p= 6.958e- 12) constructed to predict the prognosis of breast cancer. Importance analysis identifies CDKN2A has a pivotal role in AI modeling, whose higher expression indicates worse prognosis in breast cancer. Furthermore, inhibition of CDKN2A down-regulates decreases Snail1, Twist1, Zeb1, vimentin and MMP9, while E-cadherin is increased. Besides, inhibition of CDKN2A also decreases the expression of MEGEA4, phosphorylated STAT3, PD-L1, and caspase3, while cleaved-caspase3 is increased. Finally, we find down-regulation of CDKN2A or MAGEA inhibits cell migration and wound healing, respectively. AI identified CRG subgroups in pan-cancer based on CRGs-related TDGs, and 44-gene-based AI modeling is a novel tool to identify chemotherapy sensitivity in breast cancer, in which CDKN2A/MAGEA4 pathway played the most important role.

Somatic mutations in tumor-adjacent normal breast tissue in young women with high risk family history.

576 Background: It’s incompletely understood why some women without germline breast cancer predisposing mutations develop early onset breast cancer. Breast cancer in young women is often more aggressive, with high histologic grade, worse prognosis, and higher risk of recurrence. We hypothesize that these women have somatic mutations in the normal breast tissue that contribute to breast cancer tumorigenesis. Methods: Whole exome sequencing (WES) data was analyzed from normal tissue adjacent to breast tumor and compared to matched peripheral blood (n=28). All patients were &lt;50yo, had high risk family history as per NCCN guidelines, and had negative Ambry Genetics panel testing. FFPE sections were reviewed by two pathologists to identify normal breast tissue furthest away from tumor. WES was performed on Illumina HS4000 using 100-base pair paired-end reads. Samples were mapped to the human reference genome (vGRCh38). PCR duplicates were marked (Picard), and indel regions were adjusted (GATK). High-quality germline variants (HaplotypeCaller), somatic mutations (Mutect), and indels (IndelGenotyper) were called. Low-quality variants were filtered out. The classification and number of mutations per gene per sample were extracted and analyzed. Results: Majority of patients were white (86%), median age 45, with invasive ductal carcinoma (86%), ER+/PR+/HER2- (100%). Median somatic variants per sample were 97 (range: 45-285), all single nucleotide variants, mostly missense mutations, C&gt;T conversions were seen in 45% of mutations. This suggests cytidine deaminase (APOBEC) involvement. All samples had alterations in at least one of the top ten affected genes which include transcription factors in zinc finger family (listed below). Zinc finger proteins play a key role in tumorigenesis, cancer progression and metastasis in a variety of cancers. Upregulation of zinc finger proteins has been associated with worse prognosis in breast cancer. Conclusions: We describe novel somatic mutations in zinc finger proteins in normal breast tissue adjacent to tumor that may contribute to early onset breast cancer in patients with high risk family history. [Table: see text]

Abstract P2-26-06: Association between CK+/CD45+ circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) alterations in advanced breast cancer patients

Abstract Background: Circulating tumor cells (CTCs) in breast cancer (BC) are commonly defined as epithelial cells (EPCAM and cytokeratin (CK) positive), lacking the universal blood cell marker CD45. Nonetheless, CTCs expressing both CK and CD45 (= dual-positive, DP cells) can be observed in the blood of cancer patients. Early evidence suggests that DP cells might derive from the fusion of tumor cells and macrophages, and we have previously demonstrated that they present aberrant genomes and are associated with worse prognosis in BC [1,2]. Here, to further investigate the mechanisms/pathways underlying their presence, we analyzed the association between DP cells and circulating tumor DNA (ctDNA) alterations. Methods: Blood samples were collected from patients with advanced BC (aBC), before starting a new line of therapy. All patients were enrolled in a prospective clinical trial. For CTC and DP cells analysis, 7.5 ml of blood collected in CellSave® tubes was processed with the FDA-approved CellSearch® platform (positivity cutoffs were ≥1 cell for DPcells and ≥5 cells for CTCs). For ctDNA analysis, plasma was collected from Streck stabilizing tubes and analyzed with the Guardant360™ NGS platform for the detection of somatic single nucleotide variants (SNVs), insertions/deletions (indels), gene fusions/rearrangements and copy number variations (CNVs), which were then classified into pathways based on previously defined profiles generated on the Cancer Genome Atlas database (RTK, RAS, RAF, MEK, NRF2, ER, WNT, MYC, P53, cell cycle, notch, PI3K). Associations between ctDNA-detected gene alterations and circulating cell types were analyzed through chi square test, while mutant allele frequency (MAF) and number of detected alterations (NDA) were tested by Mann Whitney test. Results: We analyzed blood samples from 169 patients with luminal-like (n=80), HER2+ (n=34) and triple-negative (n=52) aBC. DPcells were detected in 85 patients (50.3 %, range 0-53), of which 40 (47 %) were CTC-positive and 45 (53%) CTC-negative. Somatic ctDNA alterations were detected in all analyzed samples. In the overall population, the presence of ≥1 DPcell was associated with SNVs in the cell cycle pathway (p = 0.043), a numerically higher incidence was also observed for CNVs in this pathway. SNVs and CNVs in the cell cycle pathway were associated with CTCs ≥ 5 as well (p = 0.005 and p = 0.003, respectively). Moreover, associations with CTCs ≥ 5 were observed for RTK SNVs and CNVs (p = 0.041 and p = 0.046, respectively), PI3K SNVs and CNVs (p = 0.006 and p = 0.007, respectively), MYC SNVs and CNVs (p = 0.042). No associations were observed in terms of MAF and NDA. In the luminal-like subgroup an association was highlighted for CNVs in the cell cycle pathway, p = 0.038. CTCs ≥ 5 were associated with PI3K SNVs (p = 0.031). In the triple-negative subgroup DPcells were associated with SNVs in the RAF pathway (p = 0.041), whereas CTCs ≥ 5 were associated with PI3K SNVs and CNVs (p = 0.044 and p = 0.024, respectively) and RTK SNVs (p = 0.008). In the HER2 positive subgroup, a higher MAF and number of detected SNVs was observed for samples with ≥1 DPcell (p = 0.0286 and p = 0.0099, respectively). Conclusions: The study analyzed ctDNA features associated with canonical and CK+/CD45+ CTCs, showing differential gene alteration profiles. Cell cycle pathway SNVs were common in both CTC populations, while other pathways (RTK, PI3K, MYC and RAF) were significantly altered in a mutually exclusive pattern. These results suggest that DPcells might have a different biological meaning compared to canonical CTCs. More studies need to be conducted to better characterize this understudied CTC subpopulation and understand their specific contribution to cancer progression.

Abstract A071: Deciphering the normal breast microenvironment in Black women and White women

Abstract Despite having similar incidence of breast cancer, black women (BW) are typically diagnosed with more aggressive subtypes of breast cancer compared to white women (WW). One mediator contributing to the disparity in breast cancer mortality rates is the distinct tumor microenvironments (TME) associated with BW and WW. Yet, characterization of the normal microenvironment (NME) in breast tissue and how they may associate with breast cancer risk factors remains unknown. Contemporary literature reports that TMEs from BW and WW with breast cancer are characterized with higher densities of pro-tumorigenic macrophages (M2) and cancer-associated adipocytes have been observed in BW compared to WW, and typically associated with worse breast cancer prognosis. While the TME has been shown to be critical for breast cancer progression, the lack of characterization of the NME is a limiting factor towards understanding the molecular differences between a normal and cancerous breast microenvironment. We hypothesize that characterizing the tissue compositions in the breast NME could elucidate unique molecular signatures that may be contributing to distinct alterations that are driving the disparity in breast cancer aggressiveness in BW. The cohorts consisted of normal breast tissue from BW (n=20) and WW (n=20) with breast cancer risk determined by Gail score. The Gail model is a well-established tool used to assess the 5-year risk of developing invasive breast cancer based on demographic and clinical data. All tissues were obtained from the Susan G. Komen tissue bank and used with institutional permission. Profiles of N-glycans, a glucose metabolism-linked post-translational modification, were characterized in normal breast tissue from BW and WW at risk for breast cancer using matrix assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI). Tissue from both cohorts were characterized based on their tissue compositions including areas rich in stroma, adipocytes, and crown like structures (CLSs), a pro-inflammatory process consisting of dying adipocytes surrounded by stroma and macrophages. Nearly 70 N-glycans were identified in the NME from both BW and WW. When comparing N-glycan peaks between premenopausal and postmenopausal women, one N-glycan peak (m/z 1905.633) was significantly higher in the premenopausal group (p&amp;lt;0.0001) regardless of genetic ancestry. However, when comparing peak m/z 1905.633 based on genetic ancestry, premenopausal and postmenopausal BW had significantly higher peak intensities compared to premenopausal and postmenopausal WW (premenopausal BW vs WW, p = 0.024; postmenopausal BW vs WW, p = 0.028). Overall, the data suggests that there may be distinct N-glycan structures associated with premenopausal and postmenopausal women based on genetic ancestry. Characterizing the NME from BW and WW at risk for breast cancer may help towards understanding potential breast stroma priming molecular signatures contributing to differences in breast cancer aggressiveness observed in BW compared to WW. Citation Format: Denys Rujchanarong, Anand Mehta, Richard R. Drake, Marvella E. Ford, Peggi M. Angel. Deciphering the normal breast microenvironment in Black women and White women [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr A071.

216P Relationship between regulatory T lymphocytes (Treg): Related genes and pathological response to neoadjuvant docetaxel-carboplatin in early-stage triple-negative breast cancer (TNBC)

An immune infiltrate rich in Treg is associated with worse prognosis in breast cancer, although there is controversy regarding their role and clinical relevance among the different tumor subtypes. This project aims to analyze the expression of Treg-related genes and its relationship with pathological response to neoadjuvant docetaxel-carboplatin in a cohort of patients with early-stage TNBC.

BREAst Cancer Personalised NuTrition (BREACPNT): dietary intervention in breast cancer survivors treated with endocrine therapy – a protocol for a randomised clinical trial

IntroductionBreast cancer survivors treated with adjuvant endocrine therapy commonly experience weight gain, which has been associated with low adherence to therapy and worse breast cancer prognosis. We aim to assess...

Racial differences in weight perception among Black and White women diagnosed with breast cancer.

Black women are more likely than White women to have obesity, and obesity is associated with worse breast cancer prognosis. Weight perception, however, has not been studied as a potential mediator of obesity disparities in women with breast cancer. In this study, we sought to describe racial differences and the association of lifestyle factors with weight perception. In this cross-sectional study design, Black and White women with a new primary breast cancer were surveyed about socio-demographics, weight perception, diet, and exercise habits.Height and weight were measured at enrollment.We classified women with a BMI ≥ 25kg/m2 or waist circumference ≥ 88cm who reported that they were "about the right weight" as under-perceivers. Chi-square and t tests were used to assess study variables (e.g., race, physical activity) associated with under-perception of weight.Logistic regression models were fit to evaluate for racial differences in under-perception while controlling for other covariates. Of 1,197women with newly diagnosed breast cancer, the average age was 58years, and 909 (75.9%) were White. Nine hundred eighteen (77%) had stage I cancer, 1,035 (87%) had estrogen receptor positive cancer, and 795 (66%) were privately insured at time of diagnosis. Seven hundred eighty-nine (66%) women had abdominal obesity (waist circumference ≥ 88cm), while 366 (31%) women had a BMI ≥ 25kg/m2.Overall, 24% of women were under-perceivers.Compared to White women, Black women with WC ≥ 88cm more frequently under-perceived their weight (24% vs. 14% p < 0.0001) were more obese with BMI > 30kg/m2 (51% vs. 23%, p < 0.0001) and had lower physical activity (22% vs. 77%, p < 0.0001). After controlling for age, education, and stage, Black women remained more likely to under-perceive their weight relative to White women for those with BMI ≥ 25kg/m2 (OR: 2.64; 95% CI: 1.4-4.6) or waist circumference ≥ 88cm (OR: 2.89; 95% CI: 1.8-4.5). With respect to lifestyle factors, among women with BMI ≥ 25kg/m2, those who met physical activity guidelines were less likely to under-perceive their weight compared to those who did not meet physical activity guidelines (OR: 0.37; 95% CI: 0.2-0.6), regardless of race. We found racial differences in weight perception and identified social determinants and lifestyle factors such as lower education and physical inactivity that influenced under-perception of weight among newly diagnosed breast cancer patients. Since obesity is associated with worse breast cancer outcomes, identifying optimal modifiable factors to intervene upon to support weight management among breast cancer survivors is clinically important. Breast cancer patients' perceptions about their weight provide insight that may inform lifestyle behavior interventions to reduce obesity during survivorship care.

Cyclooxygenase-2 immunohistochemical expression is associated with worse prognosis in breast cancer: Retrospective study and literature review.

To assess the immunohistochemistry phenotype of cyclooxygenase-2 (COX-2) in breast cancer (BC) and to correlate it with histological and clinical prognostic factors. This retrospective study utilized COX-2 monoclonal antibody in an immunohistochemistry staining of tissue microarrays slides of 570 cases of previously diagnosed BC and with 52 of normal breast tissues from breast specimens resected for benign lesions or reconstruction (fibroadenoma and normal breast epithelium). This project was carried out in the Laboratory of pathology, King Abdulaziz University, Jeddah, Saudi Arabia, between September 2019 and September 2021. The present data showed an important connection between the COX-2 expression phenotype and BC compared to benign breast tissues (p=0.034). The expression pattern of COX-2 was allied significantly with some factors which distinguished aggressive subtypes of BC, such as stage, distant metastases, lymphovascular invasion, and poor survival. Cyclooxygenase-2 is a valuable marker that could facilitate BC diagnosis and prognosis.

Adjuvant Hormone Therapy-Related Hot Flashes Predict Treatment Discontinuation and Worse Breast Cancer Prognosis.

Clinical trials have shown that adjuvant hormone therapy (AHT)-related hot flashes can predict better breast cancer outcomes. This population-based cohort study investigated whether this result can be generalized to a real-world setting. By linking the National Quality Registry for Breast Cancer, Prescribed Drug Register, and Cause-of-Death Register, we identified 7,152 chemotherapy-free patients with breast cancer who initiated AHT in Stockholm from 2006 through 2019, and followed them until 2020. Hot flashes were defined as new use of drugs for hot flashes within 6 months after initiating AHT. We used Cox models to compare disease-free survival and treatment discontinuation among patients with and without hot flashes. Patients who newly used drugs for hot flashes shortly after AHT initiation had worse disease-free survival (adjusted hazard ratio [HR], 1.67; 95% CI, 1.11-2.52) and a higher treatment discontinuation rate (adjusted HR, 1.47; 95% CI, 1.21-1.78). The association between drugs for hot flashes and discontinuation of AHT differed by patient characteristics, with stronger associations among low-income patients (HR, 1.91; 95% CI, 1.41-2.59) and those without first-degree relatives who had cancer (HR, 1.81; 95% CI, 1.39-2.35) or died from cancer (HR, 1.71; 95% CI, 1.37-2.12). AHT-related hot flashes predict worse, rather than better, breast cancer outcomes among patients in clinical routine practice. The identification of adverse effects by the initiation of hot flash medications may identify a subset of patients with more severe hot flashes who are more likely to discontinue AHT and need more support for treatment adherence.

Comprehensive Analysis of the Immune and Prognostic Implication of TRIM8 in Breast Cancer.

Background: Breast cancer remains one of most lethal illnesses and the most common malignancies among women, making it important to discover novel biomarkers and therapeutic targets for the disease. Immunotherapy has become a promising therapeutic tool for breast cancer. The role of TRIM8 in breast cancer has rarely been reported. Method: Here we identified TRIM8 expression and its potential function on survival in patients with breast cancer using TCGA (The cancer genome atlas), GEO (Gene expression omnibus) database and METABRIC (Molecular Taxonomy of Breast Cancer International Consortium). Then, TIMER and TISIDB databases were used to investigate the correlations between TRIM8 mRNA levels and immune characteristics. Using stepwise cox regression, we established an immune prognostic signature based on five differentially expression immune-related genes (DE-IRGs). Finally, a nomogram, accompanied by a calibration curve was proposed to predict 1-, 3-, and 5-year survival for breast cancer patients. Results: We found that TRIM8 expression was dramatically lower in breast cancer tissues in comparison with normal tissues. Lower TRIM8 expression was related with worse prognosis in breast cancer. TIMER and TISIDB analysis showed that there were strong correlations between TRIM8 expression and immune characteristics. The receiver operating characteristic (ROC) curve confirmed the good performance in survival prediction and showed good accuracy of the immune prognostic signature. We demonstrated the model usefulness of predictions by nomogram and calibration curves. Our findings indicated that TRIM8 might be a potential link between progression and prognosis survival of breast cancer. Conclusion: This is a comprehensive study to reveal that tripartite motif 8 (TRIM8) may serve as a potential prognostic biomarker associating with immune characteristics and provide a novel therapeutic target for the treatment of breast cancer.

The Prognostic Role of Intratumoral Stromal Content in Lobular Breast Cancer.

Simple SummaryHigh intratumoral stromal content is related to worse outcomes in several types of cancer. However, its prognostic role in breast cancer seems to differ between different subtypes. High intratumoral stromal content is a negative prognostic marker in triple-negative breast cancer, while the opposite is the case for estrogen-receptor-positive breast cancer, in which higher stromal content is indicative of a better prognosis. Most lobular breast cancers are estrogen-receptor-positive, and the tumor tissue has a clearly defined histological appearance, often with a high intratumoral stromal content. To date, the prognostic role of intratumoral stromal content in lobular breast cancer remains unclear. In this study, we aimed to investigate the prognostic importance of intratumoral stromal content in estrogen-receptor-positive lobular breast cancer. Our results show that high intratumoral stromal content is an easily assessed and clinically useful indicator of a good prognosis in lobular breast cancer.Previous studies have shown that high intratumoral stromal content is associated with a worse prognosis in breast cancer, especially in the triple-negative subtype. However, contradictory results have been reported for estrogen-receptor-positive (ER+) breast cancer, indicating that the prognostic role of intratumoral stromal content may be subtype-dependent. In this study, we investigated the importance of intratumoral stromal content for breast cancer-specific mortality (BCM) in a well-defined subgroup (n = 182) of ER+/human-epidermal growth-factor-receptor-2 negative (HER2−) invasive lobular breast cancer (ILC). The intratumoral stromal content was assessed on hematoxylin–eosin-stained whole sections and graded into high stroma (>50%) or low stroma (≤50%). A total of 82 (45%) patients had high-stroma tumors, and 100 (55%) had low-stroma tumors. High-stroma tumors were associated with a lower Nottingham histological grade, low Ki67, and a luminal A-like subtype. After a 10-year follow-up, the patients with high-stroma tumors had a lower BCM (HR: 0.43, 95% CI: 0.21–0.89, p = 0.023) in univariable analysis. Essentially the same effect was found in both the multivariable analysis (10-year follow-up) and univariable analysis (25-year follow-up), but these findings were not strictly significant. In ER+/HER2− ILC, high intratumoral stromal content is an easily assessable histological indicator of a good prognosis.

Identification of novel biomarkers in breast cancer via integrated bioinformatics analysis and experimental validation

ABSTRACT Breast cancer (BC), an extremely aggressive malignant tumor, causes a large number of deaths worldwide. In this study, we pooled profile datasets from three cohorts to illuminate the underlying key genes and pathways of BC. Expression profiles GSE42568, GSE45827, and GSE124646, including 244 BC tissues and 28 normal breast tissues, were integrated and analyzed. Differentially expressed genes (DEGs) were screened out based on these three datasets. Functional analysis including Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway were performed using The Database for Annotation, Visualization and Integrated Discovery (DAVID). Moreover, Cytoscape with Search Tool for the Retrieval of Interacting Genes (STRING) and Molecular Complex Detection (MCODE) plugin were utilized to visualize protein protein interaction (PPI) of these DEGs. The module with the highest connectivity of gene interactions was selected for further analysis. All of these hub genes had a significantly worse prognosis in BC by survival analysis. Additionally, four genes (CDK1, CDC20, AURKA, and MCM4) dramatically were enriched in oocyte meiosis and cell cycle pathways through re-analysis of DAVID. Moreover, the mRNA and protein levels of CDK1, CDC20, AURKA, and MCM4 were significantly increased in BC patients. In addition, knockdown of CDK1 and CDC20 by small interfering RNA remarkably suppressed cell migration and invasion in MCF-7 and MDA-MB-231 cells. In conclusion, our results suggested that CDK1, CDC20, AURKA, and MCM4 were reliable biomarkers of BC via bioinformatics analysis and experimental validation and may act as prospective targets for BC diagnosis and treatment.

Prognostic Value and Clinical Significance of LIPH in Breast Cancer.

BackgroundLipase member H (LIPH), a novel member of the mammalian triglyceride lipase family, is localized on human chromosome 3q27-q28. Exploration of the importance of the new cancer-related gene LIPH in several carcinomas has been reported in previous studies. Our study aims to systematically assess the expression pattern of LIPH in breast cancer.MethodsOur study explored 2994 breast cancer samples with transcriptome data from the Cancer Genome Atlas (TCGA) and the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) datasets. We systematically evaluated the mRNA expression of LIPH in breast cancer and the overall survival (OS) of patients. The protein expression of LIPH in breast cancer was evaluated with the Human Protein Atlas. We also explored the relationship between LIPH and the immune microenvironment in pan-cancer.ResultsBoth mRNA and protein expression LIPH were found to be upregulated in breast cancer tumors. The overall survival rate of patients with high LIPH expression was lower than those of patients with low LIPH expression in both the TCGA dataset (p=0.0067) and METABRIC dataset (p<0.0001). Outcomes of the multivariate analysis found that the level of LIPH expression was an independent prognostic factor in both TCGA (p=0.001) and METABRIC (p=0.019) databases. The outcomes of the univariate analysis showed that LIPH was an important prognostic factor (p=0.01 in TCGA dataset, p=0.001 in METABRIC dataset). In the TCGA dataset, outcomes showed that LIPH expression was negatively correlated with the AJCC (American Joint Committee on Cancer) stage (p=2.3e-05) and triple-negative breast cancer (TNBC) tissues (p=3.1e-10). High LIPH expression showed lower OS in the TNBC subtype (P=0.011).ConclusionCompared to normal tissues, the expression of LIPH was higher in breast cancer tissues in both mRNA and protein levels. This study showed that the high level of LIPH expression might be related to the worse prognosis of breast cancer.

Abstract 2022: Palmitate promotes breast cancer progression in vitro through induction of a senescent-like phenotype in fibroblasts

Abstract Background: Obesity confers a worse breast cancer prognosis, including an increased risk of recurrence and mortality. While the causative mechanisms have yet to be fully uncovered, emerging evidence implicates palmitate, increased in the obese state, in development of cellular senescence, an inflammatory state associated with breast tumorigenesis in preclinical models. However, studies are warranted to corroborate the impact of palmitate on induction of a cohesive senescent-like phenotype as well as the extent to which palmitate-induced senescence impacts breast tumorigenesis either in vitro or in vivo. This said, we hypothesize that palmitate exposure induces a senescent-like phenotype in fibroblasts, contributing to measures of breast cancer progression. Methods: HCA2, IMR-90, and human mammary fibroblasts were exposed to bovine serum albumin or palmitate in media supplemented with 2% charcoal-stripped fetal bovine serum, after which the cells were measured through qPCR for expression of IL-1a, IL-6 and IL-8, some of the most prominent members of the senescence-associated secretory phenotype. Palmitate-exposed fibroblasts were also stained for senescence-associated beta-galactosidase and BrdU incorporation, well-established senescence markers. Experiments were then repeated with administration of eicosapentaenoic and docosahaexanoic acids to evaluate the potential of omega-3 fatty acids to limit the effects of palmitate on fibroblast senescence. Finally, we assessed the tumor-promoting potential of these palmitate-exposed fibroblasts by culturing MCF-7 and T47D breast cancer cells in their conditioned media and assessing changes in carcinogenic measures. Results and Conclusions: Palmitate induced pro-inflammatory gene expression and SA-beta-gal positivity and decreased proliferation in fibroblasts, while omega-3 fatty acid supplementation reversed these effects. These palmitate-exposed fibroblasts also appeared to be of pathological impact, as exposure to their CM increased proliferation in breast cancer cells. These findings are important in that they support emerging evidence implicating obesity-associated factors in the exacerbation of breast cancer progression as well as indicate the potential of omega-3 fatty acids to improve outcome. Citation Format: Brittany Susanne Harlow, Albert R. Davalos, Andrew J. Brenner, Christopher Jolly, Stefano Tiziani, Stephen D. Hursting, Linda A. deGraffenried. Palmitate promotes breast cancer progression in vitro through induction of a senescent-like phenotype in fibroblasts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2022.

Race, subjective social status and metabolic syndrome in women with breast cancer.

560 Background: Metabolic syndrome (MS) is associated with worse breast cancer prognosis. Black women have higher rates of advanced breast cancer, as well as MS, diabetes and obesity. As socioeconomic status is associated with MS, we asked whether the subjective perception of social status (SSS), might influence this association in black and white women with a new breast cancer (BC) diagnosis. Methods: We surveyed, obtained serum and conducted anthropometric measures of 1206 women with a new BC diagnosis. Triglycerides, systolic blood pressure (SBP), waist circumference (cm), HDL and glucose were used to calculate a severity index of MS (MS-Zscore: mean=0; sd=1). Women reported their SSS on the 10 rung McArthur US social status ladder, their household income, education attained, diet quality (5 point scale), and exercise measured with metabolic equivalents (METS). Data were analyzed with multivariable generalized linear models. Missing data were imputed with multiple imputation. Results: Average age was 58 yrs of 295 black and 911 white women. BC stage &gt; II was in 11.6% of black and 2.4% of white women. On average black women had higher BMI (31.5 vs 26.6; p&lt;0.001), waist circumference (103 vs 93; p&lt;0.001) glucose (96 vs 92; p&lt;0.001) and SBP (132 vs 126; p&lt;0.001); lower triglycerides (92 vs 104; p&lt;0.001) and HDL (59 vs 68; p&lt;0.001). Black women were more likely than white women to live in poverty (23.7% vs 4.6%; p&lt;.001); report poor diet (32.4% vs 10.4%; p&lt;.001) and less exercise (29.7% v 23.6% in the 25th %ile), and less likely to graduate college (30.3% vs 70.4%). MSZ-score was positively associated with age (.02 per year; p&lt;.001) and black race (0.35; &lt;.001) and negatively with better diet (-.20 per point in 4 point scale); p&lt;.001), exercise (-.11 per quartile increase in METS; p&lt;.001) and SSS (-.04 per ladder rung; p=.004). SSS was lower in black women within the same levels of income and education). Conclusions: Race, age, diet, exercise and subjective social status all impact metabolic syndrome, a risk factor for breast cancer. Of concern, among breast cancer patients, black women are more likely to rate their SSS below white women, within each education or income level. Subjective social status among women with a new breast cancer diagnosis is associated with MS and may be important to address as a risk factor among breast cancer patients.[Table: see text]

Abstract PS19-13: Palmitate exacerbates breast tumorigenesis in vitro via induction of a senescent-like phenotype in fibroblasts

Abstract Background: Obesity is associated with a worse breast cancer prognosis, conferring an increased risk of recurrence and mortality. At the same time, recent evidence suggests that obesity is also correlated with development of cellular senescence, an inflammatory state associated with exacerbation of breast tumorigenesis in preclinical models. As obese individuals present greater levels of inflammation at baseline, research efforts are warranted to examine the means by which obesity promotes the development of a senescent phenotype, which may further exacerbate inflammation. Additionally, studies have yet to determine whether obesity-induced senescence modulates the tumorigenic process in the context of breast cancer specifically. Methods: To this end, we exposed fibroblasts to the obesity-associated circulating factor palmitate and used qPCR and western immunoblotting to assess the expression of genes and proteins involved in the senescent state, including interleukin (IL)-1a, IL-6, IL-8, senescence-associated beta-galactosidase (SA-beta-gal), and matrixmetalloproteinase (MMP)-9. As a mechanistic investigation, we next assessed the impact of palmitate on activation of NF-kB signaling using western immunoblotting, immunofluorescence, and NF-kB luciferase reporter assays. Finally, we utilized cell counting, MTT, wound healing, and colony formation assays to examine the impact of these palmitate-exposed fibroblasts on breast cancer cell proliferation, viability, motility, and survival, respectively. Results: We found that palmitate induced fibroblast gene expression of IL-1a, IL-6, and IL-8, major components of the senescent secretome, as well as expression of the senescent markers SA-beta-gal and MMP-9. The mechanism at least partially involved activation of NF-kB, responsible for production of about 75% of senescent secretome components. More importantly, these palmitate-exposed fibroblasts were of pathological impact, exacerbating in vitro measures of breast cancer cell aggressiveness. Conclusions: These findings contribute to our understanding of the impact of obesity-associated factors on breast tumorigenesis, demonstrating a mechanistic link between palmitate and the pro-tumorigenic effects of senescent cells. Our studies will ultimately aid in the identification of a therapeutic target that can be used to improve the comparably worse outcomes of the obese breast cancer patient. Citation Format: Brittany Susanne Harlow, Albert Davalos, Andrew Brenner, Christopher Jolly, Stefano Tiziani, Steve Hursting, Linda deGraffenried. Palmitate exacerbates breast tumorigenesis in vitro via induction of a senescent-like phenotype in fibroblasts [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 PS19-13.

ID1 and ID4 Are Biomarkers of Tumor Aggressiveness and Poor Outcome in Immunophenotypes of Breast Cancer.

Simple SummaryInhibitor of differentiation (ID) proteins are essential to promote proliferation during embryonic development, but they are silenced in most adult tissues. Evidence to date shows ID1 expression in many tumor types, including breast cancer. However, the role of the remaining ID family members, especially ID4, in breast cancer remains unclear. In this work, we aimed to assess the four ID genes expression in breast cancer cell lines and a long series of breast cancer samples and correlate them with clinicopathological features and patients’ survival. We observed a significantly higher expression of ID4 in tumor cell lines than the healthy breast epithelium cell line. We confirmed that the overexpression of ID1 and ID4 correlated with more aggressive phenotypes and poor survival in breast cancer patients’ samples. Our results support the importance of ID proteins as targets for the development of anti-cancer drugs.Inhibitor of differentiation (ID) proteins are a family of transcription factors that contribute to maintaining proliferation during embryogenesis as they avoid cell differentiation. Afterward, their expression is mainly silenced, but their reactivation and contribution to tumor development have been suggested. In breast cancer (BC), the overexpression of ID1 has been previously described. However, whether the remaining ID genes have a specific role in this neoplasia is still unclear. We studied the mRNA expression of all ID genes by q RT-PCR in BC cell lines and 307 breast carcinomas, including all BC subtypes. Our results showed that ID genes are highly expressed in all cell lines tested. However, ID4 presented higher expression in BC cell lines compared to a healthy breast epithelium cell line. In accordance, ID1 and ID4 were predominantly overexpressed in Triple-Negative and HER2-enriched samples. Moreover, high levels of both genes were associated with larger tumor size, histological grade 3, necrosis and vascular invasion, and poorer patients’ outcomes. In conclusion, ID1 and ID4 may act as biomarkers of tumor aggressiveness and worse prognosis in breast cancer, and they could be used as potential targets for new treatments discover.