Detection Of Breast Cancer Metastasis Research Articles

Abstract PS03-06: Clinical implementation of artificial-intelligence-assisted detection of breast cancer metastases in sentinel lymph nodes: saving costs and time (the CONFIDENT-B trial)

Abstract Background Metastases in sentinel lymph nodes (SN) of breast cancer (BC) patients are strongly associated with a worse survival and consequently guide treatment. If metastases are absent upon pathologist’ assessment of the regular hematoxylin and eosin (HE)-slide, additional immunohistochemistry (IHC)-stains are performed to ensure that no metastases are missed. However, these stains come at high additional costs, which may exceed specimen reimbursement. Fortunately, digital pathology is becoming more common, thereby creating an avenue of opportunities for artificial intelligence (AI) assistance. Although the number of studies on (promising) AI-algorithms increases exponentially, studies on actual clinical implementation are lacking. In this single-center prospective trial, we investigated to which extent an artificial intelligence (AI)-assisted clinical workflow for the detection of SN-metastases reduces IHC-use, while maintaining current diagnostic safety standards. Methods We enrolled 190 SN-specimens of 182 patients with invasive or in situ BC from September 2022 to May 2023. SN-specimens were allocated bi-weekly to either the control-arm (n=90) or the intervention-arm (n=100). In the control-arm, SN-specimens were digitally assessed according to the current clinical workflow, while pathologists in the intervention-arm assessed the SN-specimen with the output the ‘Metastasis-Detection-App’ (Visiopharm©) available. In both groups, IHC was performed in all morphologically negative cases. Main outcome was the relative risk (RR) of IHC-use per detected case of SN-metastases. Case-mix adjustment was performed by log-binomial regression. Results Overall, 59 SN-specimens contained metastases (31.1%). AI-assistance resulted in a significantly lower risk of IHC-use per detected case of SN-metastases (adjusted RR: 0.680, 95% CI: 0.347-0.878). Besides preventing IHC-use, thereby reducing costs, AI-assisted pathologists also spent significantly less time on their assessment of the SN-specimen (3m:41s vs. 6m:04s, p = 0.028). Furthermore, the sensitivity of AI-assisted pathologists was up to 30% higher. The AI-assisted pathologists missed two cases of micro-metastases in the intervention arm, one of which was in retrospect highlighted by the algorithm, while in the other case the tumor cells were located in a heavily cauterized area of the HE-slide and therefore only visible on the (serial) IHC-slides. In the control arm, the algorithm in retrospect picked up all micro- and macro-metastases and nearly half of the isolated tumor cells (ITC). In addition, all participating pathologists stated that AI was easy to use, that they felt confident using AI, and that besides saving them time, AI made their work more enjoyable. Cost reductions on IHC by AI-assistance depend on laboratory policy (i.e. when and on how many levels IHC is performed), but, at a cost of €25,- per IHC-stain, range from €1.500 to €3.500 per 100 SN’s in a scenario where IHC is performed in all morphologically negative cases. In a scenario where IHC is only performed in patients in whom finding ITC has clinical consequences (i.e. patients who received neoadjuvant treatment), cost savings on IHC range from €7.500-€12.500 per 100 SNs, depending on laboratory policy. Conclusion AI-implementation for the detection of SN-metastases in BC-patients leads to a significant reduction of IHC-use and subsequent costs, while saving pathologists time and making their work more enjoyable. Importantly, AI-implementation during this trial was safe and patients were not at risk of an inferior diagnosis. By doing this trial alone, an estimated €3,000 on IHC-use was saved. Such tangible cost savings are crucial to build a viable business case for AI implementation in diagnostic pathology. Citation Format: Carmen van Dooijeweert, Rachel Flach, Natalie ter Hoeve, Celien Vreuls, Roel Goldschmeding, Jan-Erik Freund, Paul Pham, Tri Nguyen, Elsken Van der Wall, Geert Frederix, Nikolas Stathonikos, Paul Van Diest. Clinical implementation of artificial-intelligence-assisted detection of breast cancer metastases in sentinel lymph nodes: saving costs and time (the CONFIDENT-B trial) [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 PS03-06.

Abstract 3789: Discovery of liquid biopsy-based novel protein biomarkers for early detection of breast cancer metastasis

Abstract Introduction and Objectives: Despite recent advances in screening and therapeutic technology, breast cancer is still the first leading cause of cancer death among females. When detected early (i.e., SEER Stage Localized), the 5-yr survival rate is 99%. The 5-yr survival rate is still 86% when cancer cells have spread beyond the breast, but this spread is limited to nearby lymph nodes (SEER Stage Regional). However, when the disease is noted to be SEER Stage Distant or metastasis, the 5-yr survival rate is significantly reduced to 31%. The evidence suggests that breast cancer is manageable if distant metastasis could be prevented. Thus, the prevailing idea is that early detection of breast cancer metastasis in surveillance will likely be the best modality to address advanced breast cancer’s dismal outcomes. Thus, liquid biopsy may be suitable for “early detection” of pre- and/or micro-metastasis because of its ease of obtaining biological material. In this pilot study, we explored the potential biomarkers for the early detection of breast cancer metastasis using publicly available datasets. Methods: We obtained 2 publicly available datasets from GEO (GSE102484 and GSE234114) containing primary tumor samples taken from subjects with and without metastatic disease. Both datasets were generated by mRNA microarray (Affymetrix Human Genome U133 Plus 2.0 Array). With these datasets, we performed the differentially expressed gene (DEG) analysis using GEO2R based on GEOquery and limma (Linear Models for Microarray Analysis) with a threshold criterion of Log 2 fold change >0 and p-value <0.05 as the cut-off point. The significantly upregulated genes were further screened using VerSeDa (Vertebrate Secretome Database) to select secreted proteins. Results: In the GSE102484 (N = 683, 582 non-metastasis and 101 metastasis), DEG identified 1798 upregulated and 1472 downregulated genes. Venn diagram analysis between the upregulated genes and VerSeDa identified 337 upregulated genes coding secreted proteins. In the GSE234114 (N = 120, 60 non-metastasis and 60 metastasis), DEG identified 2873 upregulated and 931 downregulated genes. Venn diagram analysis between the upregulated genes and VerSeDa identified 563 upregulated genes coding secreted proteins. By integrating the genes from GSE102484 and GSE234114, 82 commonly upregulated genes that code secreted proteins were identified. Conclusions: In this study, we identified 82 candidate mRNA biomarkers that are upregulated in breast tumor with metastasis. The panel of biomarkers will be validated in blood-based liquid biopsy samples in the next study. Citation Format: Cynthia Jinno, Sunao Tanaka, Takeo Fujii, Hideki Furuya. Discovery of liquid biopsy-based novel protein biomarkers for early detection of breast cancer metastasis [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 3789.

New advances in circulating tumor cell‑mediated metastasis of breast cancer (Review).

Breast cancer stands as the most prevalent form of cancer affecting women, with metastasis serving as a leading cause of mortality among patients with breast cancer. Gaining a comprehensive understanding of the metastatic mechanism in breast cancer is essential for early detection and precision treatment of the disease. Circulating tumor cells (CTCs) play a vital role in this context, representing cancer cells that detach from tumor tissues and enter the bloodstream of cancer patients. These cells travel in the blood circulation as single cells or clusters. Recent research has shed light on the enhanced metastatic potential of CTC clusters compared to single CTCs, despite their limited occurrence. The aim of the present review was to explore recent findings on CTCs with a particular focus on the clustering phenomenon of CTCs observed in breast cancer. Additionally, the present review delved into the comparison between single CTCs and CTC clusters regarding their implications for the treatment and prognosis of patients diagnosed with metastatic breast cancer. By examining the role and mechanisms of CTCs in breast cancer metastasis, the present review provided an improved understanding of CTCs and their significance in early detection of breast cancer metastasis through peripheral blood analysis. Moreover, it contributed to the comprehension of cancer prognosis and prediction by highlighting the implications of CTCs in these aspects. Ultimately, the present study seeks to advance knowledge in the field and pave the way for improved approaches to breast cancer management.

S100A14 serum level and its correlation with prognostic factors in breast cancer

BackgroundBreast cancer is the most commonly occurring cancer in women worldwide. S100A14 is a novel important member of S100 proteins family. Its importance is due to its role in tumorigenesis and metastasis process. In this study, we aimed to determine serum levels of S100A14 protein in breast cancer patients and healthy individuals to know if it can be suggested as a new biomarker for breast cancer and to reveal whether it is correlated with cancer pathological features.MethodsThis cross-sectional study was performed in two groups: study group contains 46 breast cancer patients (29 metastatic and 17 non-metastatic) and control group contains 22 healthy women. Enzyme-linked immunoabsorbent assay was performed to determine S100A14 protein levels in samples. Pathological data were obtained for each patient. The data were statistically analyzed using Kruskal-Wallis H, Mann-Whitney U, and Spearman correlation tests.ResultsS100A14 serum levels were elevated in study group compared with control group (P < 0.05). S100A14 serum levels were significantly increased in distant breast cancer patients compared with regional breast cancer patients (P = 0.001). There was a strong positive correlation between serum S100A14 level and tumor grade (rs = 0.713, P < 0.001).ConclusionOur study indicated that S100A14 serum levels are elevated in breast cancer patients compared with control individuals. High S100A14 serum levels were correlated with poor tumor differentiation so it might have a prognostic significance for breast cancer tumors. The elevation of S100A14 levels in distant breast cancer patients suggests the ability of using serum S100A14 as a biomarker for detection of breast cancer metastasis.

Accurate Detection of Breast Cancer Metastasis Using a Hybrid Model of Artificial Intelligence Algorithm

Background: Breast cancer (BC) is a prevalent disease and a major cause of mortality among women worldwide. A substantial number of BC patients experience metastasis which in turn leads to treatment failure and death. The survival rate has been significantly increased due to more rapid detection and substantial improvements in adjuvant therapies including newer chemotherapeutic and targeted agents, and better radiotherapy techniques.Methods: In this study, we cross-compared the application of advanced artificial intelligence algorithms such as Logistic Regression, K-Nearest Neighbors, Discrete Cosine Transform, Random Forest Classifier, Support Vector Machines, Multilayer Perceptron, and Ensemble to diagnose BC metastasis. We further combined MLP with genetic algorithm (GA) as a hybrid method of intelligent analysis. The core data we used for comparison belonged to the images of both benign and malignant tumors collected from Wisconsin Breast Cancer dataset from the UCI repository.Results: The application of several different algorithms to the collection of BC data indicated that these algorithms have comparable accuracy rate in detecting and predicting cancer. However, our hybrid algorithm showed superior accuracy, sensitivity and specificity compared to the individual algorithms. Two methods of comparison (Cross-Validation and Holdout) were applied to this study which produced consistent results.Conclusion: Our findings indicate that our MLP-GA hybrid algorithm can speed up diagnosis with higher accuracy rate than the individual patterns of algorithm.

Ultra-Sensitive Detection and Inhibition of the Metastasis of Breast Cancer Cells to Adjacent Lymph Nodes and Distant Organs by Using Long-Persistent Luminescence Nanoparticles

The metastasis of breast cancer mainly occurs through the axillary lymph node and blood circulation systems. It is extremely difficult to know when the cancer cells start to metastasize; however, early detection of breast cancer metastasis is crucial and challenging to enable surgical removal of the primary tumor and perform a systematic lymphadenectomy to eliminate invasion of the tumor. Herein, we report real-time tracking of the metastasis of orthotopic breast cancer with background-free near-infrared long-persistent luminescence (NIR-PL) imaging, and its guidance for the surgical removal of lymph nodes. The NIR-PL imaging is based on Cr3+/Nd3+ codoped ZnGa2O4 (A-ZGCN) nanoparticles with a superlong afterglow time of more than 15 days. We show that the detection sensitivity of metastasis of cancer cells with the NIR-PL imaging is higher than the classic bioluminescence imaging. We find that the metastasis of breast cancer cells to lymph nodes occurred as early as on the third day after orthotopic inoculation of breast cancer cells and followed an order of the proper axillary lymph node (PALN, day 3) > accessory axillary lymph node (AALN, day 6) > accessory mandibular lymph node (AMLN, day 9) > mandibular lymph node (MLN, day 25). In addition, we show that the NIR-PL nanoprobes (i.e., A-ZGCN NPs) displayed 17% radioenhancement, which was used for radiotherapy of orthotopic breast cancer to further prevent and reduce its metastasis to other organs. The radiotherapy treatment is superior to surgery for removal of the tumor accompanied by a NIR-PL imaging-guided lymphadenectomy. Our work demonstrates the great potential of NIR-PL imaging and the corresponding nanoprobes for tracking metastasis of cancer cells and for radiotherapy.

LRP6 Ectodomain Prevents SDF-1/CXCR4-Induced Breast Cancer Metastasis to Lung.

Lung metastasis is an important cause of breast cancer-related deaths, in which SDF-1/CXCR4 signaling pathway plays a critical role. Single transmembrane protein LRP6 is viewed as an oncogene via activating the Wnt/β-catenin signaling pathway. Our work aims to investigate the relationship between SDF-1/CXCR4 and LRP6 in breast cancer lung metastasis. We examined the expressions and functions of SDF-1/CXCR4 and LRP6 as well as their relationship in breast cancer in vitro and in vivo. LRP6 ectodomain (LRP6N) directly bound to CXCR4 and competitively prevented SDF-1 binding to CXCR4. LRP6N prevented SDF-1/CXCR4-induced metastasis to lung and prolonged survival in mice bearing breast tumors, whereas LRP6 knockdown activated SDF-1/CXCR4 signal transduction and promoted lung metastasis and tumor death. Furthermore, patients with breast cancer with high CXCR4 expression had poor prognosis, which was exacerbated by low LRP6 expression but improved by high LRP6 expression. Interestingly, a secreted LRP6N was found in the serum of mice and humans, which was downregulated by the onset of cancer metastasis in both mice bearing breast cancer as well as in patients with breast cancer. LRP6N might be a promising diagnostic marker for the early detection of breast cancer metastasis as well as an inhibitor of SDF-1/CXCR4-induced breast cancer metastasis. LRP6N also provides an interesting link between Wnt signaling and SDF-1/CXCR4 signaling, the two key pathways involved in cancer development.

Abstract 3560: MicroRNA-665 promotes metastasis of breast cancer by targeting nuclear receptor subfamily 4 group A member 3

Abstract Background: It is the result of metastasis to the lymph nodes or other organs that leads breast cancer (BC) patients to death. Then the early detection of metastasis in BC is helpful to regulate progression of BC and then predict prognosis of BC patients. Since the abnormal level of miRNAs are bound up with BC and metastasis, of which may act as biomarkers to predict prognosis of BC, we find that expression level of miR-665 is ectopic up-regulated in BC patients and it may serve as an estimable biomarker of prognosis of BC patients. Methods: The expressions of miRNA were detected by our current microarray system and verified by quantitative real-time reverse transcription-PCR (qRT-PCR). In Student’s t-test and Kaplan-Meier analysis, the levels of miR-665 were positively correlated with poor survival in BC patients. Multivariate survival analysis identified either miR-665 expression or TNM stage as an independent prognostic factor of BC. Diverse roles of miR-665 played in various cellular functions were examined by loss-of-function and gain-of-function effects in vitro and in vivo. Dual-luciferase reporter gene experiment was then used to confirm the direct complementation between miR-665 and NR4A3, accompanied by activating the epithelial-mesenchymal transition (EMT). Furthermore, siRNAs targeting NR4A3 were used to rescue cell migration and invasion in miR-665 knockdown cells. Results: Expression of miR-665 was upregulated which predicted poor prognosis in BC patients. Since miR-665 may regulate key processes of tumorigenesis, targeting of NR4A3 was proved to promote cell migration and invasion by activating the EMT. Accordingly, either silencing of miR-665 or over-expressing of NR4A3 rescued the migration and invasion phenotype in vitro. Conclusions: Our study suggests that miR-665 plays an important part in progression of BC and promotes cell migration as well as invasion by down-regulation of NR4A3. The data collectively indicate that miR-665 is an onco-miRNA and may represents a potential predictor for diagnosis and prognosis of BC. Citation Format: Xinge Zhao. MicroRNA-665 promotes metastasis of breast cancer by targeting nuclear receptor subfamily 4 group A member 3 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3560.

Abstract 23: Chemotherapy and tau interplay facilitates breast-to-brain metastasis

Abstract Breast cancer represents the second most common cause of brain metastasis with the rate of 10-16% after lung cancer. The overall survival in brain metastatic patients is very short ranging from 2 to 25 months. Considering the high incidence of brain metastasis and the limitation of available therapies, developing preventive strategies and early detection methods are required to improve the survival rate in the patients. Many primary and brain metastatic breast cancer patients represent cognitive deficits. Besides, chemo-treated breast cancer patients show memory problems before brain metastases diagnosis. Although the cause of cognitive impairments after chemotherapy have not been explored, a recent study revealed that chemotherapy promotes breast cancer metastasis. Correspondingly, our mRNA expression analysis showed downregulation of the tight junction markers in chemo-treated choroid plexus cells (CPs), suggesting that chemotherapy is likely to cause an increase in the blood-cerebral spinal fluid barrier (BCSFB) permeability. Chemotherapy also induces upregulation of tau expression in CP cells. Tau is known as a pathological hallmark of Alzheimer's disease (AD) and neurodegenerative disorders. The abnormally phosphorylated and aggregated form of tau play critical role in neurodegenerative diseases as of AD. Tau expression, both at the mRNA and protein level, is significantly elevated in breast to brain metastases (BBMs). Therefore, we hypothesized that chemotherapy upregulates tau expression in breast cancer cells leading to the BCSFB and blood-brain barrier (BBB) permeability, and consequently, breast to brain metastasis and neurodegeneration. To test our hypothesis, the effect of chemotherapy on the BCSFB and BBB permeability were investigated using the trans endothelial electrical resistance (TEER) and migration assays in the in vitro models. Our results show an increase in abnormal tau expression in chemo-treated primary breast cancer cells. BBM-derived tau is abnormal and forms paired helical filaments (PHFs), similar to Alzheimer's. We conclude that breast to brain metastasis is facilitated through an interplay between chemotherapy, and tau expression and release from breast cancer cells. Additionally, we suggest that tau plays a key role in neurodegeneration defects in brain metastatic patients. This study opens a new path towards prevention and early detection of breast cancer metastasis to the brain, cognitive decline and neurodegeneration in metastatic patients. Citation Format: Behnaz Saatian, Krutika Deshpande, Alex Julian, Brooke Naomi Nakamura, Ling Shao, Josh Neman. Chemotherapy and tau interplay facilitates breast-to-brain metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 23.

MicroRNA-200c and microRNA- 141 are regulated by a FOXP3-KAT2B axis and associated with tumor metastasis in breast cancer

BackgroundMembers of the microRNA (miR)-200 family, which are involved in tumor metastasis, have potential as cancer biomarkers, but their regulatory mechanisms remain elusive.MethodsWe investigated FOXP3-inducible breast cancer cells, Foxp3 heterozygous Scurfy mutant (Foxp3sf/+) female mice, and patients with breast cancer for characterization of the formation and regulation of the miR-200 family in breast cancer cells and circulation. Participants (259), including patients with breast cancer or benign breast tumors, members of breast cancer families, and healthy controls, were assessed for tumor and circulating levels of the miR-200 family.ResultsFirst, we identified a FOXP3-KAT2B-miR-200c/141 axis in breast cancer cells. Second, aging Foxp3sf/+ female mice developed spontaneous breast cancers and lung metastases. Levels of miR-200c and miR-141 were lower in Foxp3sf/+ tumor cells than in normal breast epithelial cells, but plasma levels of miR-200c and miR-141 in the Foxp3sf/+ mice increased during tumor progression and metastasis. Third, in patients with breast cancer, the levels of miR-200c and 141 were lower in FOXP3low relative to those with FOXP3high breast cancer cells, especially in late-stage and metastatic cancer cells. The levels of miR-200c and miR-141 were higher in plasma from patients with metastatic breast cancer than in plasma from those with localized breast cancer, with benign breast tumors, with a family history of breast cancer, or from healthy controls. Finally, in Foxp3sf/+ mice, plasma miR-200c and miR-141 appeared to be released from tumor cells.ConclusionsmiR-200c and miR-141 are regulated by a FOXP3-KAT2B axis in breast cancer cells, and circulating levels of miR-200c and miR-141 are potential biomarkers for early detection of breast cancer metastases.

First experience of lymphoscintigraphy use in early stages of breast cancer

Aim. To evaluate efficacy of radioisotopic methods in detection of metastatic involvement of sentinel lymph nodes for choosing further theurapeutic tactics. &#x0D; Methods. The study group included 23 females with confirmed breast cancer. Sentinel lymph nodes were detected by isotope 99mTc Nanocolloid and blue dye in General Electric gamma camera (USA). &#x0D; Results. In all cases patients’ age was 45 to 60 years. In 16 (69.5%) cases the tumor size was below 5 cm which corresponded to clinical stage 2, in other 7 (30.5%) patients the tumor size was below 2 cm. Positive results with the use of lymphoscintigraphy were revealed in 6 (26%) cases, all results were morphologically confirmed straight after lymph node removal. In 5 (83%) cases metastatic involvement of the node was observed in stage 2 cancer. Such patients were immediately referred to radical mastectomy with radical lymph node dissection. In other cases pathological accumulation of a radiopharmaceutical was not observed. Only in one case after removal of such node its metastatic involvement was confirmed histologically. So the frequency of false negative results was 4.3%. Respectively, status of sentinel lymph nodes in breast cancer reflects the status of lymphatic collector with 95.7% accuracy. &#x0D; Conclusion. The biopsy results demonstrated high diagnostic value of lymphoscintigraphy for detection of breast cancer metastases to sentinel lymph nodes (sensitivity 86%, specificity 100%): the method can be successfully used in early stages of breast cancer and also in case of negative results of conventional diagnostic methods (ultrasound, roentgen mammography).

Mathematical and Statistic Method to Diagnostic of Metastasis based on Radiology Images for Breast Cancer

: One out of eight women face with the problem of breast cancer in her life. It is the second causes of women death in the world. Detection of breast cancer metastasis at the earliest stage is important to predict and control of breast cancer progression. It can spread to the different parts of the body, such as are the bone, liver, and lung and etc. How and where does the breast metastasis grow and migrate may open many questions to the radiologist and medical Scientifics. In this study, we discussed about the possibility of the breast metastasis to the other parts of the body and using statistic method we are able to estimate the frequency of metastasis from breast to the other parts. We have selected statistic method to this research and we studied over 200 patients (effected by breast cancer) using radiology image data. We save the history of the patient and gather all patient data in a table. Result and discussion: the result show that the most metastasis of breast occur in the brain and the less occur in liver. The result may help radiologist to understand better origin of metastasis and in case the possibility is high, the radiologist may ask for breast imaging. A New discussion comes to the mind that, do we need to perform breast imaging if radiologist faced with metastasis on brain?

Encapsulated Conjugated Oligomer Nanoparticles for Real-Time Photoacoustic Sentinel Lymph Node Imaging and Targeted Photothermal Therapy.

Noninvasive and nonionizing imaging of sentinel lymph nodes (SLN) is highly desirable for the detection of breast cancer metastasis through sentinel lymph node biopsy. Photoacoustic (PA) imaging is an emerging imaging technique that can serve as a suitable approach for SLN imaging. Herein, novel conjugated oligomer based nanoparticles (NPs) with strong NIR absorption, good biocompatibility, excellent PA contrast, and good photothermal conversion efficiency are reported. Real-time PA imaging of SLN reveals high resolution of the NPs via injection from the left forepaw pad. In addition, the surface functionalized NPs can target breast cancer cells and kill them efficiently and specifically through photothermal therapy upon 808 nm laser irradiation. This work shows great potential of the nanoparticle PA contrast agent to serve as a multifunctional probe for photothermal therapy at SLNs to achieve the inhibition of cancer cell metastasis in the near future.

Possibilities of positron emission tomography in the metastatic breast cancer diagnosis

An analysis of published data on the possibilities of the positron emission tomography method application in complex diagnostic and treatment activities in metastatic breast cancer was performed. The possibilities are described and the method characteristics according to series of studies, in particular with regard to the various localization metastases identification are given. It is noted that, compared with conventional imaging techniques, positron emission tomography is more sensitive and specific method of distant metastases diagnosis. The results of clinical studies on assessment of the method application possibilities in the detection of breast cancer metastases in lymph nodes, various organs and bone metastases are given. It has been shown that the introduction of sodium fluoride into practice allowed to use this agent as bone tissue biomarker, which can be used in positron emission tomography performing. The data that the simultaneous use of positron emission tomography with 18F-fluorodeoxyglucose and computed tomography has a high diagnostic value in relation to the breast cancer distant metastases detection are presented. The method possibilities for the various forms of primary invasive breast cancer identification, including invasive ductal and lobular cancer are considered. The prospects analysis for a further increase of opportunities for positron emission tomography application is performed, in particular, reports on the possibility of so-called hybrid scanning using positron emission tomography and magnetic resonance imaging are considered. It is highlighted that the most important aim of the method use in clinical practice in the prospect may be local-regional recurrence and distant metastasis monitoring after primary treatment for breast cancer.

Abstract 3973: Comparison of different highly sensitive miRNA array platforms for the investigation of circulating cell-free microRNAs in blood

Abstract Introduction: Circulating free (cf) microRNAs hold great promises in early detection and prognosis of human diseases such as cancer. However due to the very low amount of cf miRNA in plasma and serum, highly informative miRNAs are lost. Thus, there is an urgent need for highly efficient, sensitive, cost efficient technologies to measure low abundant miRNA in a quantitative manner. Previously, we have successfully measured and validated miRNA profiles of early onset breast cancer cases, healthy controls and breast cancer patients who had developed breast cancer metastasis during follow-up by applying TaqMan Low density arrays. We established and validated miRNA signatures for the early detection of early stage breast cancer, and prognosis and early detection of breast cancer metastasis. However more sensitive microarray platforms with highly dynamic ranges could even improve identified diagnostic signatures especially in respect of early detection of diseases and relapse. Here, we compared the sensitivity of the previously applied array platform TaqMan Low density array (TLDA) with Toray innovative 3D-Gene microarray technology by measuring plasma cf miRNA profiles of 40 individuals (10 healthy controls, 10 early stage breast cancer, 20 metastatic breast cancer patients). Experimental Procedures: Plasma samples were processed within 2 h after blood collection. EDTA blood was centrifuged for 15 min at 1200 g and the supernatant centrifuged again at 12.000 g for 10 min in order to remove any cell debris. Cf RNA was isolated from plasma. Cf miRNA profiles of plasma samples of 40 individuals (10 healthy controls, 10 early stage breast cancer, 20 metastatic breast cancer patients) were investigated A: via TaqMan low density arrays (TLDA) and B: via 3D-Gene microarray technology. Expression profiles between both array types and between cases, early stage breast cancer and metastatic breast cancer patients were compared. In addition selected miRNAs were measured with single TaqMan assays. These results were compared with the array based results. Conclusion: The result will be of importance for all scientists who intend to develop and improve diagnostic miRNA panels. Based on our results recommendations for a microarray platform for the investigation of cf miRNA will be given. Citation Format: Melanie Maierthaler, Eric Mennesson, Katarina Cuk, Dharanija Madhavan, Manuela Zucknick, Isabelle Fixe, Alexandra Foucher, Markus Wallwiener, Andreas Schneeweiss, Nadia Normand, Barbara Burwinkel. Comparison of different highly sensitive miRNA array platforms for the investigation of circulating cell-free microRNAs in blood. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3973. doi:10.1158/1538-7445.AM2015-3973

In vitro selection of DNA aptamers for metastatic breast cancer cell recognition and tissue imaging.

Cancer is a major public health issue, with metastatic cancer accounting for the overwhelming majority of cancer deaths. Early diagnosis and appropriate treatment of metastatic cancer may largely prolong the survival rate and improve the quality of life for patients. In this study, we have identified a panel of DNA aptamers specifically binding to MDA-MB-231 cells derived from metastatic site-pleural effusion, with high affinity after 15 rounds of selections using the cell-based systematic evolution of ligands by exponential enrichment (SELEX) method. The selected aptamers were subjected to flow cytometry and laser confocal fluorescence microscopy to evaluate their binding affinity and selectivity. The aptamer LXL-1 with the highest abundance in the enriched library demonstrated a low K(d) value and excellent selectivity for the recognition of the metastatic breast cancer cells. Tissue imaging results showed that truncated aptamer sequence LXL-1-A was highly specific to the corresponding tumor tissue and displayed 76% detection rate against breast cancer tissue with metastasis in regional lymph nodes. Therefore, on the basis of its excellent targeting properties and functional versatility, LXL-1-A holds great potential to be used as a molecular imaging probe for the detection of breast cancer metastasis. Our result clearly demonstrates that metastatic-cell-based SELEX can be used to generate DNA ligands specifically recognizing metastatic cancer cells, which is of great significance for metastatic cancer diagnosis and treatment.

SWIFT MRI enhances detection of breast cancer metastasis to the lung.

To evaluate the capability of longitudinal MR scans using sweep imaging with Fourier transformation (SWIFT) to detect breast cancer metastasis to the lung in mice. Mice with breast cancer metastatic to the lung were generated by tail vein injection of MDA-MB-231-LM2 cells. Thereafter, MR imaging was performed every week using three different pulse sequences: SWIFT [echo time (TE) ∼3 μs], concurrent dephasing and excitation (CODE; TE ∼300 μs), and three-dimensional (3D) gradient echo (GRE; TE = 2.2 ms). Motion during the long SWIFT MR scans was compensated for by rigid-body motion correction. Maximum intensity projection (MIP) images were generated to visualize changes in lung vascular structures during the development and growth of metastases. SWIFT MRI was more sensitive to signals from the lung parenchyma than CODE or 3D GRE MRI. Metastatic tumor growth in the lungs induced a progressive increase in intensity of parenchymal signals in SWIFT images. MIP images from SWIFT clearly visualized lung vascular structures and their disruption due to progression of breast cancer metastases in the lung. SWIFT MRI's sensitivity to fast-decaying signals and tolerance of magnetic susceptibility enhances its effectiveness at detecting structural changes in lung parenchyma and vasculature due to breast cancer metastases in the lung.

Molecular detection of breast cancer metastasis in sentinel lymph nodes by reverse transcriptase polymerase chain reaction (RT-PCR): identifying, evaluating and establishing multi-marker panels

The potential advantage of using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) methodology to detect metastasis in sentinel lymph nodes (SLNs) of breast cancer (BC) patients was evaluated in this prospective study. We measured the expression of relevant gene transcripts in SLNs using an innovative algorithm and compared the results of single-marker assays versus multi-marker assays with conventional histological detection methods. SLNs from women aged ≥ 18 years diagnosed with unilateral BC were examined by haematoxylin-eosin staining and immunohistochemistry and analysed for transcripts of several relevant genes using qRT-PCR (learning group). Four candidate panels of expressed transcript combinations with high sensitivity and specificity were selected for further investigation. The candidate panels were then validated using SLNs from a second group of BC patients (validation group). In the learning group, 74/314 SLN sections from 150 patients were positive for metastasis by histology. The transcripts analysed showed the following individual sensitivities/specificities: cytokeratin 19 (CK19) 94.6%/97.9%; mammaglobin 1 (MGB1) 82.4%/91.7%; mammaglobin 2 (MGB2) 82.4%/96.7%; carcinoembryonic antigen (CEA) 71.6%/97.5%; EPCAM (epithelial cell adhesion molecule) 91.9%/97.1%; and NY-BR-1 82.4%/93.8%. The optimal panel based on the predefined criteria comprised four markers: CK19, MGB1, EPCAM, and NY-BR-1, of which ≥ 2 had to be positive (95.9% sensitivity, 95.0% specificity, 85.5% positive predictive value (PPV), and 98.7% negative predictive value (NPV)). Overall concordance with histology was 95.2%. In the validation group, 84/315 SLN sections from 235 patients were histologically positive, and panel sensitivity, specificity and overall accuracy were 88.1, 95.2 and 93.3%, respectively, at the SLN section level. In conclusion, molecular staging using expression patterns of relevant transcripts in SLNs could serve as a useful complement to standard diagnostic work-up in BC patients. The proposed flexible multi-parametric approach does not improve the overall accuracy compared with the single-marker approach. However, it overcomes several limitations of the previously reported molecular assays for SLN diagnosis.

A mammaglobin-A targeting agent for noninvasive detection of breast cancer metastasis in lymph nodes.

Pathologic axillary lymph node (ALN) status is an important prognostic factor for staging breast cancer. Currently, status is determined by histopathology following surgical excision of sentinel lymph node(s), which is an invasive, time consuming, and costly procedure with potential morbidity to the patient. Here, we describe an imaging platform for noninvasive assessment of ALN status, eliminating the need for surgical examination of patients to rule out nodal involvement. A targeted imaging probe (MamAb-680) was developed by conjugation of a mammaglobin-A-specific monoclonal antibody to a near-infrared fluorescent dye. Using DNA and tissue microarray, mammaglobin-A was validated as a cell-surface target that is expressed in ALN-positive patient samples but is not expressed in normal lymph nodes. In vivo selectivity was determined by i.v. injection of MamAb-680 into mice with mammaglobin-A-positive and -negative mammary fat pad (MFP) tumors; and by peritumoral MFP injection of the targeted imaging probe in mice with spontaneous ALN metastases. Fluorescence imaging showed that probe was only retained in positive tumors and metastases. As few as 1,000 cells that endogenously express mammaglobin-A were detected in ALN, indicating high sensitivity of this method. Translation of this approach offers considerable potential as a noninvasive clinical strategy to stage breast cancer.

A study of the assessment of axillary lymph nodes before surgery for breast cancer using multidetector-row computed tomography

Sentinel lymph node biopsy (SLNB) is widely used in the detection of breast cancer metastasis, and a retrospective study was conducted to determine whether the preoperative assessment of axillary lymph node metastasis using multidetector-row computed tomography (MDCT) images would contribute to the selection of patients who require SLNB. Seventy of the 164 patients who underwent surgery of the breast during the period of April 2006 to December 2008 were selected as subjects who: (1) did not undergo preoperative chemotherapy; (2) had undergone MDCT before the surgery; (3) had undergone either SLNB or axillary lymph node removal during the operation; and (4) whose T factor was pathologically T2 or less. The diameter and shape of the largest lymph node that was identified on an MDCT image of the axilla on the affected side were measured and compared with permanent pathological specimens. Size: The group with lymph node metastasis included 21 subjects with an average diameter of the largest lymph node of 10.3 mm (range, 6-23 mm), and the nonmetastasis group included 49 subjects with that of 7.1 mm (5-13 mm). Shape: Round lymph nodes that internally contained no fat concentration in 24 subjects, and cuneate or round lymph nodes that internally contained a fat concentration in 46 subjects. The observable lymph nodes that were round had a diameter of at least 10 mm, and internally contained no fat concentration (A) were regarded as positive, whereas the cuneate or round lymph nodes that measured less than 8 mm in diameter and internally contained fat concentration (B) were regarded as negative, and both positive predictive value and negative predictive value was 100%. Axilla removal was performed from the beginning in the case of (A) by assessing the size and shape for the presence or absence of axillary lymph node metastasis before surgery using MDCT.