Late-stage Breast Cancer Patients Research Articles

SMARCD1 is an essential expression-restricted metastasis modifier

Breast cancer is the most frequently diagnosed cancer worldwide, constituting 15% of cases in 2023. The predominant cause of breast cancer-related mortality is metastasis, and a lack of metastasis-targeted therapies perpetuates dismal outcomes for late-stage patients. By using meiotic genetics to study inherited transcriptional network regulation, we have identified, to the best of our knowledge, a new class of “essential expression-restricted” genes as potential candidates for metastasis-targeted therapeutics. Building upon previous work implicating the CCR4-NOT RNA deadenylase complex in metastasis, we demonstrate that RNA-binding proteins NANOS1, PUM2, and CPSF4 also regulate metastatic potential. Using various models and clinical data, we pinpoint Smarcd1 mRNA as a target of all three RNA-BPs. Strikingly, both high and low expression of Smarcd1 correlate with positive clinical outcomes, while intermediate expression significantly reduces the probability of survival. Applying the theory of “essential genes” from evolution, we identify 50 additional genes that require precise expression levels for metastasis to occur. Specifically, small perturbations in Smarcd1 expression significantly reduce metastasis in mouse models and alter splicing programs relevant to the ER+/HER2-enriched breast cancer. Identification subtype-specific essential expression-restricted metastasis modifiers introduces a novel class of genes that, when therapeutically “nudged” in either direction, may significantly improve late-stage breast cancer patients.

Analysis of Presentation Delay in Patients with Advanced Breast Cancer Visiting Dharmais National Cancer Hospital

Background: Breast cancer is still one of the significant causes of the increasing mortality rate due to cancer. Treatment of breast cancer patients was delayed by 66.34%. The majority of patients arrive at stages IIIB and IV and are therefore treated at an advanced stage. This study aimed to analyze the factors causing the arrival delay of breast cancer patients to Dharmais Cancer Hospital with advanced stages. Methods: A descriptive study with a qualitative approach was used as the research design. Indepth interviews and document reviews were used to collect data. In-depth interviews were conducted with 12 advanced-stage breast cancer patients. Thematic data analysis was performed by tracing and identifying the main topics of the entire interview process. The triangulation method was used to validate the data by conducting interviews with 4 patient families and 2 oncology surgeons, as well as document review. Results: The presentation delay of advanced breast cancer patients was analyzed based on the interpretation and recognition of symptoms. There was an attitude of ignoring when there were symptoms and not seeking treatment. On psychological factors, there was a feeling of fear after being diagnosed and a sense of worry in the treatment process. On socioeconomic factors, there was no early detection, lack of information on early detection, waiting time constraints, and the selection of alternative treatments. Conclusions: Late-stage breast cancer patients were drawn to Dharmais Cancer Hospital due to issues with the interpretation and experience of symptoms, psychological issues patients reported, and socioeconomic difficulties during treatment. As a result, patients must be given additional care to deal with late arrivals, especially in cases of advanced breast cancer.

Abstract 3692: Genomic profiling with the novel single-cell IMPACT assay reveals evidence of genetic heterogeneity among CTCs in late-stage breast cancer patients

Abstract Introduction: Liquid biopsy enables minimally invasive molecular profiling of systemic cancer for diagnosis, therapy selection and longitudinal monitoring of minimal residual disease. Cell-based liquid biopsy methods are particularly well suited to examine the extent of genetic heterogeneity and clonal diversity among metastatic cancer cells, which are believed to be the major determinants of therapeutic failure. However, isolation and molecular profiling of circulating tumor cells (CTCs) from body fluids is challenging due to methodological constrains. Methods: To overcome these challenges, we have developed a workflow for detection, isolation, and mutational profiling of CTCs. The workflow utilizes the FDA-approved CellSearch System® and a novel method for single-cell Integrated Mutation Profiling of Actionable Cancer Targets (scIMPACT). Our workflow was optimized for reliable detection of copy number variations (CNVs) and single nucleotide variations (SNVs) in single CTCs. The method was then applied to 31 CTCs alongside with 15 matched germline samples. Results: After implementation of a customized Bayesian neural network based algorithm, accounting for biases introduced during amplification of single-cell DNA and prioritizing the detected mutations, we achieved a sensitivity of 91% and a specificity of 96%, AUC = 0.98. The CTC analysis revealed presence of CNVs affecting known oncogenes and tumor suppressor genes (including MYC, CCND1, FGF4, TP53 and RB1) as well as SNVs classified as pathogenic (e.g., ARID1A Q775*). Notably, in the examined specimens most of the chromosomal aberrations (51-82%) were clonal across all cells of a given patient. In contrast, most of the detected somatic SNVs (67-96%) were sub-clonal and thus present only in a subset of CTCs. Conclusion: In summary, the novel scIMPACT workflow enabled reliable and accurate genomic profiling of patient-derived CTCs in late-stage breast cancer patients. Our proof-of-concept study revealed evidence of genetic diversity of SNVs among the examined CTCs, which might have been acquired late in progression but relevant for therapy escape in the studied cases. Further analyses are still needed for assessing the generalizability of our finding as well as for deeper understanding of the molecular basis orchestrating the establishment and maintenance of heterogeneous genotypes of CTCs in breast cancer. Citation Format: Zbigniew T. Czyż, Clara Chaiban, Adithi Ravikumar Varadarajan, Cäcilia Köstler, Vadim Dechand, Jonas Grote, Thomas Ragg, Bernhard Polzer, Jens Warfsmann, Christoph A. Klein. Genomic profiling with the novel single-cell IMPACT assay reveals evidence of genetic heterogeneity among CTCs in late-stage breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3692.

Abstract 4796: Improving cancer screening performance for women with dense breast tissue

Abstract Breast cancer is the most common cancer in women in the United States. To reduce mortality, screening by mammography in females is recommended by the United States Preventive Services Task Force, successfully reducing mortality by 15 - 32% in women aged 40 - 69 years. This is partially due to the high compliance rate (60 - 80%) among the candidate population. Mammography increases early-stage disease detection, which is correlated with better outcomes. While mammography has been largely successful, this screening method has limitations. Overall, mammography has false-negative and false-positive rates estimated at 12 - 20% and 7 - 12%, respectively. Consequently, annual mammograms result in 50 - 60% of women having a false-positive result within a 10-year span. Additionally, high breast density, found in approximately 50% of females over the age of 40, increases susceptibility to false positive and negative results. For this reason, in 2023 the FDA required that dense breast tissue status, an independent risk factor for cancer, be disclosed to mammogram recipients. The subsequent diagnostic pathway for women with dense breast tissue is not well defined for the care provider. Alternative imaging options have performance issues or are less accessible. Here we describe a liquid biopsy test to complement mammography in women with dense breast tissue to help resolve ambiguity in results. Blood from early- and late-stage breast cancer patients was prospectively collected in Streck cfDNA BCT Devices (n=100) or retrospectively acquired (n=100); and blood from presumed normal samples was collected (n=200). cfDNA was extracted and converted into libraries for low-pass whole genome sequencing (LP-WGS). Sequenced reads were analyzed to generate fragment end-motif and size (FEMS) and fragment coverage dataframes for classification using Genece Health’s proprietary analysis pipeline and machine-learning (ML) algorithm. 150 breast cancer samples distributed across stages and 150 presumed normal samples were selected to create a training cohort. Performance was tested using a 5-fold cross validation strategy. The best performance was observed using a convolution neural network (CNN) ML model with >80% sensitivity when specificity was set at 85%. The remaining 50 breast cancer samples and 50 presumed normal samples were used as an external test set. The performance in this cohort was comparable to the training cohort, with both specificity and sensitivity >80%. Genece presents an early proof of concept breast cancer screening liquid biopsy assay that uses LP-WGS fragmentomics and ML to detect cancer from a single blood sample. Continuing development with additional cohorts and optimization will yield even greater performance. The Genece liquid biopsy test has the potential to be paired with mammography, especially in women with dense breast tissue, to improve screening sensitivity, specificity, and overall outcomes. Citation Format: Molly Smith, Mengchi Wang, Andrew Carson, Byung In Lee, Michael Salmans, Bryan Leatham. Improving cancer screening performance for women with dense breast tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4796.

Immune-nutritional indices in female Nigerian breast cancer patients with different tumour characteristics

Breast cancer presents varied and complex clinicopathological features which make its management individualistic, prolonged and expensive, thus the need to differentiate each type of breast cancer using cheap and easy to perform investigations. This study evaluated the differentiation potential of extensive immune-nutritional indices which combined serum C-reactive protein (CRP), haemoglobin, albumin levels, blood lymphocyte and platelet counts in breast cancer patients of different status. The aim of this study was to investigate whether pre-treatment Onodera’s prognostic nutritional index (OPNI), CRP-Albumin-Lymphocyte (CALLY) index and the Haemoglobin, Albumin, Lymphocyte and Platelet (HALP) scores differ with breast cancer characteristics. We enrolled a total of 135 consecutive newly diagnosed breast cancer patients who were not on any treatment from Radiation Oncology Department, University College Hospital, Ibadan, Nigeria. Lymphocyte count, platelet count and haemoglobin levels were determined using auto-analyser while serum CRP and albumin levels were measured using ELISA. Immune-nutritional indices (OPNI, CALLY and HALP) were calculated. Haemoglobin concentration, serum albumin level, OPNI, CALLY and HALP were significantly decreased while serum CRP was significantly raised in breast cancer patients compared with control. Mean values of OPNI, CALLY and HALP were significantly decreased in late stage breast cancer patients compared with early stage breast cancer patients, while only CALLY was significantly decreased in poorly differentiated and metastatic breast cancer patients compared with well differentiated and non-metastatic breast cancer patients respectively. This study confirmed the presence of malnutrition and inflammation in breast cancer patients, especially late-stage, poorly differentiated and metastatic groups. This calls for monitoring of liver functions in all cases of breast cancer.

Haematological and biochemical analysis of blood samples from early and late stage breast cancer patients in India.

Breast cancer is the most prevalent cancer with the maximum number of cases worldwide. Early diagnosis of the cancer is necessary for an effective treatment plan. Due to a lack of awareness, diagnosis of breast cancer at an early stage is difficult. The present study aims to evaluate and compare the haematological and biochemical profiles of the early and late-stage breast cancer patient's data records. A retrospective cohort study was conducted on 56 breast cancer patients at the Institute of Medical Sciences, Banaras Hindu University India. Patient data records were obtained and haematological and biochemical parameters were arranged on an Excel sheet and analyzed. Random blood sugar (RBS), alkaline phosphates (ALP) levels, and urea levels were significantly high in patients with late-stage breast cancer (Tumor stage III and IV). At the advanced stage of breast cancer hemoglobin level falls and patients became anemic. Further large-scale studies with a greater number of patient data can help establish these parameters individually or in combination as prognostic and diagnostic markers in breast cancer staging.

The intention of utilization and experience toward traditional Chinese medicine among breast cancer patients in the early and late stages: a qualitative study

BackgroundIn Taiwan, breast cancer patients usually take conventional medicine and traditional Chinese medicine simultaneously. The utilization of traditional Chinese medicine among breast cancer patients at various stages has not been examined. This study aims to compare the intention of utilization and experience toward traditional Chinese medicine among early- and late-stage breast cancer patients.MethodThis qualitative research collected data from breast cancer patients through focus groups interview by convenience sampling. Conducted in 2 branches of Taipei City Hospital, a public hospital managed by the Taipei City government. Breast cancer patients > 20 years old and had used TCM for breast cancer therapy for at least 3 months were included in the interview. A semi-structured interview guide was adopted in each focus group interview. In the following data analysis, stages I and II were considered early-stage, and stages III and IV were late-stage. For analyzing the data and reporting the results, we used qualitative content analysis as the approach for data analysis, assisted by NVivo 12. Categories and subcategories were identified through content analysis.ResultsTwelve and seven early- and late-stage breast cancer patients were included in this study, respectively. The side effects were the main intention of utilizing traditional Chinese medicine. Improving side effects and constitution was the main benefit for patients in both stages. Additionally, early-stage breast cancer patients used traditional Chinese medicine to prevent recurrence or metastasis. Late-stage breast cancer patients responded more frequently to the use of traditional Chinese medicine due to the side effects of western medicine. However, some of their symptoms were not fully relieved.ConclusionsBreast cancer staging may influence the intention and utilization of traditional Chinese medicine. Health policymakers should consider the results of this research and the evidence-based illustrations to establish guidelines for integrating traditional Chinese medicine among various stages of breast cancer to improve the outcome and quality of care for cancer patients.

PCR220 Symptom Burden in Late-Stage Breast Cancer Patients Treated with CDK Inhibitors Using the PRO-CTCAE

Cyclin-Dependent Kinase Inhibitors (CDKI) are an important advancement in the treatment of Metastatic Breast Cancer. Barriers to clinician adoption, such as tolerability concerns, may limit front line use. The objective of this study is to use electronic patient-reported outcome (ePRO) and remote symptom monitoring platform to evaluate symptom differences among late-stage breast cancer patients with and without CDKI treatment.

Predictive biomarker for cisplatin in prospective phase 2 of liposomal cisplatin in metastatic breast cancer.

3130 Background: Heavily pretreated late-stage breast cancer patients of any subtype represent a huge treatment challenge. Cisplatin may work in selected patients but is often disqualified because of toxicity. LiPlaCis is a liposomal formulation of cisplatin in development utilizing the sPLA2-triggered targeted release of cisplatin. The prediction of cisplatin efficacy with a 205 mRNA profile has previously been demonstrated in a retrospective study in two NSCLC cancer patient cohorts receiving adjuvant cisplatin1. Methods: The dose-escalation part of a phase 1/2 study, of 20 patients with a variety of locally advanced or metastatic solid tumors, suggested a phase 2 dose of LiPlaCis to be 75mg on day 1 and 8 q 3 wks. In this phase 2 part, the cisplatin drug response predictor (DRP) was used to select metastatic breast cancer (mBC) patients with response likelihood scores from FFPE diagnostic or later biopsies. Patients were treated until progression or unacceptable toxicity. Halfway through phase 2 part, the dosing was changed to a per body surface schedule (40 mg/m2) after a fatal event with the flat dosing (in effect 51 mg/m2 for this patient). Among the 52 patients included in the phase 2 part, we here report data from the 37 patients with mBC platinum naïve, with DRP scores 34-100%, and with RECIST-defined response outcomes. Within this subgroup, we conducted a post hoc analysis of possible DRP cut-off values that could identify patients with a clinically meaningful response to treatment. Results: mBC patients had received a median of 7 previous treatment lines. A DRP score ≥80% (DRP80+) discriminated well between responders and non-responders to liposomal cisplatin. All 4 partial remissions in the study were in the DRP80+ group, and other key efficacy endpoints were in favor of the DRP80+ vs the lower scores (DRP80-). LiPlaCis was generally well tolerated, with a mean of 4.4 treatment cycles administered per patient. Only 1 SAE (pyelonephritis) was reported and considered related to treatment. One patient died whilst on treatment, but due to disease progression. From a total of 164 treatment cycles, 18 of the patients reported 1 grade 4 and 41 grade 3 AEs considered possibly related to treatment. Conclusions: LiPlaCis shows efficacy in heavily pretreated mBC patients, especially when using the DRP method as a companion diagnostic. Further, this is the first clinical study to prospectively validate DRP as a method to identify responders to chemotherapy cisplatin. 1) Buhl et al PLOS One doi: 10.1371/journal.pone0194609. Clinical trial information: NCT01861496 . [Table: see text]

Novel diagnostic biomarkers of T cell-mediated tumor killing characteristics for early-stage triple negative breast cancer: A SEER analysis and molecular portraits.

The primary objective was to investigate the epidemiology, molecular characteristics, and clinical survival to identify potential transcriptome biomarkers to promote early diagnosis and screening of triple-negative breast cancer patients. Early-stage triple-negative breast cancer patients (E-TNBC) and late-stage triple-negative breast cancer patients (L-TNBC) were identified from the Surveillance, Epidemiology, and End Results database from 2010 to 2019. The difference in cancer specific survival (CSS) and overall survival (OS) between E-TNBC and L-TNBC was analyzed via a Kaplan-Meier plotter. 118 triple-negative breast cancer samples and 114 normal samples with the RNA sequencing expression data were selected from the cohort of TCGA breast cancer from UCSC Xena Database. The study involved 13,690 patients with L-TNBC and 44,994 patients with E-TNBC. L-TNBC patients were more frequently to be ≤ 60 years old (54.9% vs 52.2%), multiple primary site (44.0% vs 30.1%), and were more likely to receive radiotherapy (49.6% vs 47.4%) and chemotherapy (81.1% vs 72.1%), while L-TNBC patients were less likely to be white (68.7% vs 73.0%), married or with domestic partner (46.7% vs 54.7%), poorly differentiated grade (54.0% vs 61.9%), < 3 months from diagnosis to treatment (91.6% vs 96.4%), and were less likely to receive surgery (72.3% vs 95.4%). Stage-stratified survival analysis revealed that the prognosis of L-TNBC was worse when compared to E-TNBC, Kaplan-Meier analysis demonstrated that there were striking differences in OS and CSS between E-TNBC and L-TNBC. In the multivariable regression models, L-TNBC was the single highest risk factor, with a death risk that was 4.741 and 6.074 times higher than E-TNBC in terms of OS and CSS, respectively. The results also showed that treatment with surgery, radiation, or chemotherapy was effective for a better prognosis. Transcriptome analyses revealed that the top 5 upregulated genes in L-TNBC were, respectively, ISX, ALOX15B, MADCAM1, TP63, and ARG1 compared with E-TNBC. And the top 5 downregulated genes were, respectively, CTAG1B, CT45A1, MAGEC2, TFF2, and TNFRSF11B. The L-TNBC exhibited a lower rate of survival than E-TNBC, and the 2 groups differed in terms of transcriptome characteristics. To date, the diagnostic value of T cell-mediated tumor killing portraits on E-TNBC may not be completely recognized.

Abstract 3678: Drivers of racial, regional, and socioeconomic disparities in metastatic breast cancer mortality

Abstract Despite an overall decline in breast cancer (BC) mortality due to advancements in cancer therapy, mortality disparities by race, rurality, and socioeconomic status (SES) persist among women diagnosed with metastatic disease in the US. Women residing in high-poverty or rural areas or who are non-Hispanic Black (NHB) experience higher rates of BC mortality relative to their counterparts. Although mortality disparities among late-stage BC patients are well-documented, few studies have examined the drivers of these disparities, which are likely multifactorial. We sought to identify tumor, treatment, and patient characteristics that may contribute to differences in BC mortality by race, rurality, and SES among women diagnosed with a first primary stage IIIB - IV BC in Georgia.Using the Georgia Cancer Registry, we identified 3085 patients with an initial diagnosis of stage IIIB-IV primary BC between January 2013 and December 2017. Cox proportional hazards regression was used to calculate the hazard ratios (HRs) and 95% confidence intervals (CIs) to compare NHB vs. non-Hispanic White (NHW), rural vs. urban residents, and residents of low- vs. high-SES neighborhoods by tumor (stage, grade, ER status, and molecular subtype), treatment (surgery type, receipt of chemotherapy, radiation, hormonal, neoadjuvant, and immunotherapy), and patient (race, insurance, age group, marital status, region, SES) characteristics. Using an extension of the counterfactual framework, we estimated the mediating effects of subtype, stage, SES, rurality, and insurance on the association between race and BC mortality. Among the study population, 41% were NHB, 21% resided in rural counties, and 72% resided in low SES neighborhoods. Overall, we observed mortality disparities by race (HR=1.27, 95% CI: 1.13, 1.41) and rurality (HR=1.14, 95% CI: 1.00, 1.30), but not by SES (HR=1.04, 95% CI: 0.91, 1.19). In the stratified analyses, racial disparities were the most pronounced among women with HER2 overexpressing tumors (HR=2.30, 95% CI: 1.53, 3.45). Residing in a rural neighborhood was associated with increased mortality among uninsured women (HR=2.25, 95% CI: 1.31, 3.86) or receipt of breast-conserving surgery (HR=2.21, 95 CI%: 1.32, 3.71). The most pronounced socioeconomic disparities were among younger women (&amp;lt;40 years: HR=1.46, 95% CI: 0.88, 2.42) and patients who received neoadjuvant therapy (HR=1.44, 95% CI: 1.01, 2.05). The mediation analysis demonstrated that 48% of the effect between race and BC mortality was mediated by subtype.There is considerable variation in racial, regional, and socioeconomic disparities in metastatic BC mortality by tumor, treatment, and patient characteristics. For each, we’ve identified patient groups where disparities are most pronounced. Understanding specific barriers within these patient groups will inform future interventions aimed at reducing disparities in metastatic BC mortality. Citation Format: Leah Moubadder, Lindsay J. Collin, Rebecca Nash, Jeffrey Switchenko, Jasmine Miller-Kleinhenz, Keerthi Gogineni, Kevin C. Ward, Lauren E. McCullough. Drivers of racial, regional, and socioeconomic disparities in metastatic breast cancer mortality [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3678.

Breast Cancer in Indonesia in 2022: 30 Years of Marching in Place

Breast Cancer in Indonesia in 2022: 30 Years of Marching in Place

Liquid Biopsy and Tissue Biopsy Comparison with Digital PCR and IHC/FISH for HER2 Amplification Detection in Breast Cancer Patients.

Two hundred twenty-four breast cancer patients with paired tissue and plasma samples were enrolled from 3 clinical centers to evaluate sensitivity and specificity of a digital PCR HER2 amplification assay. All patients were histologically confirmed diagnosis of locally advanced and recurrent or metastatic breast cancer with stage III/IV and had tissue HER2 status determinations using IHC/FISH. For the whole 224 advanced breast cancer patients, the sensitivity between dPCR in plasma and IHC/FISH in tissue samples is 43.75% (42/96), the specificity is 84.38% (108/128) and the overall concordance is 66.96% (150/224). Interestingly, when we looked at stage III, stage IV and recurrent or metastatic breast cancer separately, compared with IHC/FISH in tissue samples, the sensitivity of dPCR in plasma increases from 37.93% (11/29) for stage III to 41.67% (15/36) for stage IV cancer. Recurrent breast cancer patient had an increased sensitivity of 51.61% (16/31). This is consistent with our expectation sensitivity would increase concordantly as tumor burden goes up. On the other hand, specificity decreased from 92.68% (38/41) for stage III to 86.44% (51/59) for stage IV cancer. Recurrent breast cancer patient had a specificity of only 67.86% (19/28). This is, in part, due to inter- and intra-tumor heterogeneity. Many patients determined to be negative for HER2 amplification in tissue biopsy could have HER2 positive tumors at other sites, which was detected by the liquid biopsy. This study suggested the necessity of liquid biopsy for HER2 amplification detection and demonstrated digital PCR can be used as a companion diagnostic tool to determine HER2 amplification status. It also suggested that a liquid biopsy should follow a negative result from tissue biopsy to avoid false negative results especially for late-stage breast cancer patients and ones who experienced relapse or became resistant to current therapy. Future studies should focus on therapeutic effects on patients determined to be HER2 positive through liquid biopsy and collecting additional tissue biopsies to identify HER2 positive tumor when the original tissue biopsy and liquid biopsy don't agree.

Quality of Life in Palliative Post-mastectomy Reconstruction: Keystone versus Rotational Flap.

Background:Late stage breast cancer presents with malignant wound causing skin infiltration, pain, bleeding, and malodour, which affect quality of life (QoL). Palliative mastectomy aims to eliminate wound symptoms and requires prolonged wound care to improve QoL. This study aimed to prospectively investigate QoL differences in 2 alternative reconstructive methods: keystone flap and rotational flap.Methods:Twenty-four late stage breast cancer patients with symptoms of cancer wounds were included in this study. They were divided into 2 groups: keystone flap and rotational flap. Each patient’s QoL was evaluated using EORTC QLQ-C30 and QLQ-BR23 before and 3 weeks after surgery.Results:Global health post-surgery was significantly improved compared with pre-surgery in all patients (P < 0.001), across both the keystone (P = 0.018) and rotational groups (P = 0.007). Breast symptoms post-surgery were also improved compared with pre-surgery in all patients (P = 0.035). However, when analyzed per group, breast symptoms were only improved significantly in the keystone group (P = 0.013) but not in the rotational group (P = 0.575). When compared between 2 groups, future perspective post-surgery in the keystone group [100 (0–100)] was better than the rotational group [66.7 (0–100)], (P = 0.020).Conclusions:Reconstructive surgery after mastectomy improves QoL in late stage breast cancer patients. The keystone flap is superior to the rotational flap in improving global health and breast symptoms.

Analysis of CK5/6 and EGFR and Its Effect on Prognosis of Triple Negative Breast Cancer.

BackgroundTriple-negative breast cancer (TNBC) is considered to be higher grade, more aggressive and have a poorer prognosis than other types of breast cancer. Discover biomarkers in TNBC for risk stratification and treatments that improve prognosis are in dire need.MethodsClinical data of 195 patients with triple negative breast cancer confirmed by pathological examination and received neoadjuvant chemotherapy (NAC) were collected. The expression levels of EGFR and CK5/6 were measured before and after NAC, and the relationship between EGFR and CK5/6 expression and its effect on prognosis of chemotherapy was analyzed.ResultsThe overall response rate (ORR) was 86.2% and the pathological complete remission rate (pCR) was 29.2%. Univariate and multivariate logistic regression analysis showed that cT (clinical Tumor stages) stage was an independent factor affecting chemotherapy outcome. Multivariate Cox regression analysis showed pCR, chemotherapy effect, ypT, ypN, histological grades, and post- NAC expression of CK5/6 significantly affected prognosis. The prognosis of CK5/6-positive patients after NAC was worse than that of CK5/6-negative patients (p=0.036). Changes in CK5/6 and EGFR expression did not significantly affect the effect of chemotherapy, but changes from positive to negative expression of these two markers are associated with a tendency to improve prognosis.ConclusionFor late-stage triple negative breast cancer patients receiving NAC, patients who achieved pCR had a better prognosis than those with non- pCR. Patients with the change in expression of EGFR and CK5/6 from positive to negative after neoadjuvant chemotherapy predicted a better prognosis than the change from negative to positive group.

High-Resolution Patterning of Hydrogel Sensing Motifs within Fibrous Substrates for Sensitive and Multiplexed Detection of Biomarkers.

There has been an increasing and urgent demand to develop nucleic acid bioassays which not only offer high analytical performance but which are also amenable with point-of-care testing. Hydrogels present a versatile class of materials with biocompatible antifouling properties and the ability to be engineered for a range of advanced sensing applications. Fibrous substrates like nitrocellulose offer low-cost and durable platforms to run complex bioassays while enabling portability and ease of handling. We demonstrate herein the ability to synergistically combine these two materials into a portable biosensing platform by leveraging projection lithography. We demonstrate the direct polymerization of hydrogel sensing motifs within a range of fibrous substrates with precise control over their shape, size, location, and functionality. Spatial encoding of the hydrogel motifs enables the multiplex detection of multiple biomarkers on the same test. As a proof-of-concept, we apply the platform to the detection of microRNA, an emerging class of circulating biomarkers with promising potential for early diagnosis and monitoring of cancer. The assay offers a large dynamic range (over three orders of magnitude), high sensitivity (limit of detection of 2.5 amol), as well as versatility and ease of handling. Finally, the bioassay is validated using real biological samples, namely, total RNA extracted from the sera of late-stage breast cancer patients, demonstrating its utility and compatibility with clinical biosensing applications.

High PKCλ expression is required for ALDH1-positive cancer stem cell function and indicates a poor clinical outcome in late-stage breast cancer patients.

Despite development of markers for identification of cancer stem cells, the mechanism underlying the survival and division of cancer stem cells in breast cancer remains unclear. Here we report that PKCλ expression was enriched in basal-like breast cancer, among breast cancer subtypes, and was correlated with ALDH1A3 expression (p = 0.016, χ2-test). Late stage breast cancer patients expressing PKCλhigh and ALDH1A3high had poorer disease-specific survival than those expressing PKCλlow and ALDH1A3low (p = 0.018, log rank test for Kaplan-Meier survival curves: hazard ratio 2.58, 95% CI 1.24–5.37, p = 0.011, multivariate Cox regression analysis). Functional inhibition of PKCλ through siRNA-mediated knockdown or CRISPR-Cas9-mediated knockout in ALDH1high MDA-MB 157 and MDA-MB 468 basal-like breast cancer cells led to increases in the numbers of trypan blue-positive and active-caspase 3-positive cells, as well as suppression of tumor-sphere formation and cell migration. Furthermore, the amount of CASP3 and PARP mRNA and the level of cleaved caspase-3 protein were enhanced in PKCλ-deficient ALDH1high cells. An Apoptosis inhibitor (z-VAD-FMK) suppressed the enhancement of cell death as well as the levels of cleaved caspase-3 protein in PKCλ deficient ALDH1high cells. It also altered the asymmetric/symmetric distribution ratio of ALDH1A3 protein. In addition, PKCλ knockdown led to increases in cellular ROS levels in ALDH1high cells. These results suggest that PKCλ is essential for cancer cell survival and migration, tumorigenesis, the asymmetric distribution of ALDH1A3 protein among cancer cells, and the maintenance of low ROS levels in ALDH1-positive breast cancer stem cells. This makes it a key contributor to the poorer prognosis seen in late-stage breast cancer patients.

Abstract P1-09-07: Sub-group of PIK3CA WT breast cancer patients have hyperactive S1P signaling tumor cells responsive to PIK3-inhibitors: Functional signaling profile test identifies new patient group who may benefit from PIK3-targeted therapies

Abstract Background: Less than 20% of PI3KCA mutated late stage breast cancer patients achieved an objective response in a Phase III clinical trial with alpelisib, a recently approved PIK3CA inhibitor. This suggests biological factors other than PIK3CA status, such as aberrant GPCR-linked signaling, may be important to measure when identifying patients eligible for PI3K inhibitors. A new assay using an impedance biosensor was developed to measure GPCR-initiated signaling activity and the involvement of the PI3K node in transducing this activity in live tumor cells. The CELx PI3K Signaling Function (CELx PI3K) Test measures ex vivo live tumor cell response in real-time to a specific S1P agonist and PI3K antagonists to diagnose breast tumors with PI3K-involved hyperactive signaling. This study set out to: 1) characterize the involvement of the PI3K node in hyperactive S1P-initiated signaling in patient breast cancer cells and cell lines; and 2) assess whether PI3K-involved hyperactive S1P signaling is limited to breast cancer cells with PI3KCA mutations. Methods: A panel of 18 fresh HER2-/PIK3CA WT tumor cells from breast cancer patients and five PIK3CA mutated breast tumor cell lines were obtained. Real-time live cell response to an S1P agonist, an alpha-specific PI3K antagonist (alpelisib), a gamma-specific PI3K antagonist (IPI-549), and a pan-PI3K inhibitor (taselisib) were measured using an xCELLigence RTCA impedance biosensor. From these responses, SIP-initiated signaling was quantified and a previously determined cutpoint was used to identify abnormal levels of signaling activity. The relative and net amount of PI3K isoform participation in S1P-initiated signaling was also quantified. Results: Five of the 18 PIK3CA WT primary patient breast tumor cells and two of the five PIK3CA mutated breast tumor cell lines (BT20 and HCC1954) had abnormal levels of S1P-initiated signaling activity. The mean reduction of abnormal SIP signaling in the five primary breast tumor cells with IPI-549 was two times greater than with alpelisib (60% vs. 30%). In the two PIK3CA mutated cell lines with abnormal S1P-initiated signaling, the mean S1P signaling reduction with IPI-549 and alpelisib was not significant (0% and 28%), but the mean S1P signaling reduction with taselisib was 82%. Conclusions: The CELx PI3K test found hyperactive S1P-initiated signaling involves the PI3K node in PI3KCA mutated and WT breast tumor cells. The abnormal levels of PI3K-involved signaling in PIK3CA WT patient breast cancer tumors were comparable in magnitude to levels found in well-characterized PI3KCA mutated breast tumor cell lines. Unlike the PIK3CA mutated cells lines, the bulk of the PIK3-involved signaling in the PIK3CA WT patient tumor cells was associated with the PI3K-gamma isoform rather than the PI3K-alpha isoform. The superior reduction of S1P signaling by taselisib vs. alpelisib in the HCC1954 cell line (74% vs. 21%) as determined with the CELx PI3K test is consistent with previously reported xenograft studies that found taselisib induced reduction of HCC1954 tumors whereas alpelisib had no significant effect. This study thus suggests that measurement of PI3K-involvement in hyperactive S1P signaling in live patient breast cancer cells may provide a means to identify breast cancer patients who may benefit from treatment with PI3K inhibitors. Citation Format: Lance Laing, Salmaan Khan, Ian MacNeil, Catherine Kuzmicki, Samantha Kharbush, Benjamin Rich, Abhay Shukla Shukla, Kelly Brass Brass, Brian Sullivan Sullivan. Sub-group of PIK3CA WT breast cancer patients have hyperactive S1P signaling tumor cells responsive to PIK3-inhibitors: Functional signaling profile test identifies new patient group who may benefit from PIK3-targeted therapies [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P1-09-07.

Abstract P6-14-13: New approach to nipple reconstruction: In vivo evaluation of acellular nipple-areolar complex grafts

Abstract There are over 3M breast cancer survivors in the US, many of whom have undergone breast reconstructive surgery. With one-third of early stage and two-thirds of late-stage breast cancer patients opting for mastectomies without a standard, viable method to reliably reconstruct a permanent and natural nipple-areolar complex (NAC). NAC reconstruction is of key psychological importance for these patients, decreasing feelings of distress and increasing body image and self-esteem. Current NAC reconstruction options include rubber prostheses, 3D tattoos, and surgical techniques to create NAC-like structures from a patient’s own skin tissue. These approaches produce NACs that either lack physical depth or fail to maintain a protrusion. The objectives of the non-human primate 6 week engraftment studies with our decellularized human nipple areolar complex (hNAC) grafts were 1) to determine the biocompatibility of the hNAC graft and 2) to measure host-mediated recellularization of the hNAC graft on a nearest-to-human model species. If the hNAC graft maintains cell-derived insoluble matrix components yet is devoid of host cells, intracellular material, and pathogenic organisms, then the hNAC graft will not elicit an adverse humoral response and will show both good biocompatibility and recellularization. The methods employed for our studies involved surgical engraftment of the hNAC graft along the dorsal midline of three adult male non-human primates (rhesus macaque). The number of hNAC grafts engrafted for each NHP was 12 for one NHP and 18 for the other two. Gross animal weight, complete blood count data, and metabolite data were collected weekly from periods before, during (span of 6-weeks), and after hNAC engraftment for each animal. In addition, hNACs were surgically removed at defined weekly time points during the study for histological analysis of recellularization. All data was compiled for each study and statistically analyzed separately and combined. Statistical analyses included unpaired, parametric comparisons of systemic indicators (i.e. weight, complete blood count, metabolites) at each period of the study and of re-epithelialization and neovascular content for individual hNAC grafts during host-mediated recellularization over time. The results from this NHP study demonstrate good evidence of biocompatibility of the hNAC on non-human primates. Gross body weight remained unchanged for all NHPs and complete blood counts (e.g. red blood cells, white blood cells, platelets) and metabolites remained mostly within normal ranges for all NHPs. The hNAC grafts did not effect liver, kidney, or pancreas function. The hNAC grafts showed good recellularization during the study with significant increases measured for re-epithelialization and neovascularization. Histological analysis further revealed functioning microvasculature within grafts as early as 3-weeks post-engraftment. These data suggest that the hNAC graft is biocompatible and capable of recellularization. The presence of de novo epithelia and vasculature within the hNAC grafts collectively support recellularization and regeneration of a hNAC. Citation Format: David Graham, William Heim, Vincent Caronna, Abigail Chaffin, Brooke Grasperge, Scott Sullivan, Bruce A Bunnell, Nicholas C Pashos. New approach to nipple reconstruction: In vivo evaluation of acellular nipple-areolar complex grafts [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P6-14-13.

MicroRNA-429 inhibits bone metastasis in breast cancer by regulating CrkL and MMP-9.

Bone metastasis is common in late-stage breast cancer patients and leads to skeletal-related events that affect the quality of life and decrease survival. Numerous miRNAs have been confirmed to be involved in metastatic breast cancer, such as the miR200 family. Our previous study identified microRNA-429 (miR-429) as a regulatory molecule in breast cancer bone metastasis. However, the effects of miR-429 and its regulatory axis in the metastatic breast cancer bone microenvironment have not been thoroughly investigated. We observed a positive correlation between miR-429 expression in clinical tissues and the bone metastasis-free interval and a negative correlation between miR-429 expression and the degree of bone metastasis. We cultured bone metastatic MDA-MB-231 cells and used conditioned medium (CM) to detect the effect of miR-429 on osteoblast and osteoclast cells in vitro. We constructed an orthotopic bone destruction model and a left ventricle implantation model to examine the effect of miR-429 on the metastatic bone environment in vivo. The transfection experiments showed that the expression levels of V-crk sarcoma virus CT10 oncogene homolog-like (CrkL) and MMP-9 were negatively regulated by miR-429. The in vitro coculture experiments showed that miR-429 promoted osteoblast differentiation and that CrkL promoted osteoclast differentiation. The two animal models showed that miR-429 diminished local bone destruction and distant bone metastasis but CrkL enhanced these effects. Furthermore, CrkL and MMP-9 expression decreased simultaneously in response to increased miR-429 expression. These findings further reveal the possible mechanism and effect of the miR-429/CrkL/MMP-9 regulatory axis in the bone microenvironment in breast cancer bone metastasis.