Benign Breast Disease Research Articles

Non-invasive screening and subtyping for breast cancer by serum SERS combined with LGB-DNN algorithms

Early detection of breast cancer and its molecular subtyping is crucial for guiding clinical treatment and improving survival rate. Current diagnostic methods for breast cancer are invasive, time consuming and complicated. In this work, an optical detection method integrating surface-enhanced Raman spectroscopy (SERS) technology with feature selection and deep learning algorithm was developed for identifying serum components and building diagnostic model, with the aim of efficient and accurate noninvasive screening of breast cancer. First, the high quality of serum SERS spectra from breast cancer (BC), breast benign disease (BBD) patients and healthy controls (HC) were obtained. Chi-square tests were conducted to exclude confounding factors, enhancing the reliability of the study. Then, LightGBM (LGB) algorithm was used as the base model to retain useful features to significantly improve classification performance. The DNN algorithm was trained through backpropagation, adjusting the weights and biases between neurons to improve the network's predictive ability. In comparison to traditional machine learning algorithms, this method provided more accurate information for breast cancer classification, with classification accuracies of 91.38 % for BC and BBD, and 96.40 % for BC, BBD, and HC. Furthermore, the accuracies of 90.11 % for HR+/HR- and 88.89 % for HER2+/HER2- can be reached when evaluating BC patients' molecular subtypes. These results demonstrate that serum SERS combined with powerful LGB-DNN algorithm would provide a supplementary method for clinical breast cancer screening.

Coupling of the expression of proliferation and epithelialmesenchymal transition markers with the histidine-rich glycoprotein HRG mRNA expression in breast diseases

Non-malignant breast diseases (NMBD) may increase the risk of developing a malignant neoplasm. Therefore, it seems relevant to search for criteria for cell malignancy in NMBD. Aim of the study was to investigate the relationship between expression of proliferation and epithelial-mesenchymal transition (EMT) markers and histidine-rich glycoprotein (HRG) mRNA in breast diseases. Material and methods. In breast biopsy specimens of 37 patients with invasive carcinoma of a non-specific type (ICNT) and 17 patients with NMBD expression of proliferation markers (Ki-67, cyclin D1 (CCND1)) and EMT markers (E-cadherin (CDH1), type II collagen (CII) and β1-integrin (CD29)) was determined immunohistochemically. HRG mRNA expression was estimated using real time PCR. Results. HRG mRNA expression was detected in 91.9 % cases (34 of 37) in ICNT, 82.4 % (14 of 17) in NMBD and in the latter case was inversely related to the expression of CDH1, CD29 and Ki-67. A direct relationship has been established between the presence of Ki-67 and CCND1, CII, between CCND1 and CD29 in NMBD. In patients with ICNT, a direct correlation was found between the HRG mRNA expression and the presence of CII, and an inverse correlation between the number of cells containing CII and CD29. It was found that in ICNT and NMBD with the presence of HRG mRNA expression, the CDH1 expression is less than in its absence. Conclusions. Indicators of HRG mRNA expression in NMBD, combined with the assessment of proliferation and EMT markers, can be useful in developing criteria for cell malignancy in benign breast diseases.

Role of Sonoelastography in Differentiating Benign and Malignant Breast Lesions: A Prospective Study Comparing Elasticity Contrast Index and Tsukuba Score

Introduction: Breast cancer is the most common cancer occurring in women globally which is potentially curable if detected early. Ultrasound elastography is a dynamic technique that estimates tissue stiffness to differentiate between benign and malignant masses. Our study aimed to determine and compare the diagnostic accuracy of B mode Sonography, Elasticity contrast index, and Tsukuba score in differentiating malignant and benign breast masses. Methods: This was a prospective cross-sectional study done including 110 lesions in 102 patients in the age group of 15-73 years. The solid breast lesions seen on sonography were categorized according to the American College of Radiology Breast Imaging Reporting and Data System (ACR BI-RADS) and further evaluated with elastography using both Elasticity Contrast Index (ECI) and Tsukuba score with pathological diagnosis taken as the gold standard. The cut-off value of ECI was obtained. The diagnostic accuracy of B-mode sonography, ECI, and Tsukuba score was compared. Results: We found that B-mode sonography had a sensitivity of 85.7%, specificity of 100%, and accuracy of 96.9 %. The accuracy of Tsukuba scores for differentiating benign and malignant lesions was 81.2%. The cut-off value of ECI obtained was 2.8 and the accuracy was 81.8%. A statistically significant correlation (p < 0.05) existed between sonographic diagnosis, ECI, and Tsukuba score. Conclusions: B mode sonography had the highest diagnostic accuracy while ECI and Tsukuba scores were comparable. ECI can be used for breast masses using a cut-off of 2.8 to differentiate benign from malignant.

Shear wave elastography of the breast‐histopathological comparisons

AbstractIntroductionThis study aimed to compare shear wave elastography (SWE) of benign and malignant breast lesions, evaluate the sensitivity and specificity of previously suggested thresholds for identifying malignant breast lesions, and compare SWE measures across histopathological type and grade.MethodsThis single‐centre study included 303 patients, and 405 solid breast lesions were biopsied and evaluated by mammography, which may have included contrast‐enhanced mammography, conventional B‐mode ultrasound, and SWE. Following this, elastography mean (Emean), maximum (Emax), and ratio (Eratio) variables were calculated for elasticity in kilopascals (kPa) and speed in metres per second (m/s).ResultsMalignant (n = 113) samples were significantly higher than benign (n = 267) across all SWE variables [median (interquartile range)]; Emean (kPa) [132.8 (34), vs. 41.9 (53.6), p < .001], Emean (m/s) [6.9 (1), vs. 3.7 (2.2), p < .001]. The highest combined sensitivity and specificity were for Emax (97%, 73%) at >80 kPa, and Emean (96%, 76%) at >5.2 m/s. Histopathological grade 3 (kPa), Emean [145.8 (22.5), p = .012], Emax [147.8 (19.4), p = .009], and Eratio [11.7 (6.6), p = .006] were significantly higher than grades 1 [129.2 (37.3), 133.2 (35.9), 7.53 (4.7)] and 2 [127.3 (36.4), 133.3 (34.2), 8.3 (6.1)].ConclusionBreast elastography is a sensitive complementary technique that can distinguish between malignant and benign lesions and help characterise histological profile and grade.

OA15 A case series of idiopathic granulomatous mastitis patients and their immunosuppressive management

Abstract Background/Aims Rheumatologists are experts in managing immunosuppression and are asked to supervise immunosuppression for non-rheumatic conditions. Thus, it is important to be aware of these conditions and the regimes required. Idiopathic granulomatosis mastitis (IGM) is a rare inflammatory breast condition which can have systemic manifestations including arthritis. Methods Our index case presented with peripheral ankle synovitis/tenosynovitis, struggling to weight bear, whilst being investigated for a breast mass. Breast US, mammogram and biopsy (acute and chronic inflammatory cells including multi-nucleated giant cells) were completed and there was little response to 3 courses of antibiotics. No mycobacteria were seen on microscopy, ZN stain negative, CXR normal, T spot negative and serum ACE normal. There was no rash and no conditions associated with a seronegative spondyloarthropathy. RF, anti-CCP and ANA were negative. A moderate course of prednisolone immediately resolved the peripheral arthritis, but high-dose steroids were required to reduce the breast mass. The mass recurred upon weaning prednisolone requiring the addition of azathioprine as a steroid-sparing agent. Results Our case series includes the immunosuppressive management of 3 other IGM patients: 1) 43-year-old referred after an 8 year history of alternating breast ‘abscesses’ and sinuses treated with courses of antibiotics to little avail. High dose prednisolone and methotrexate resolved the breast mass within a month. 2) 37-year-old referred with a 14 year history of breast masses and fistulae previously treated with surgery and antibiotics. Currently asymptomatic so commenced on methotrexate. 3) 39-year-old referred with a 3 year history of recurrent sterile breast abscesses previously managed with antibiotics, incision and drainage. Recent small masses had settled with NSAIDs. Conclusion IGM is a rare benign inflammatory breast disease of unknown aetiology, commonest in Middle Eastern populations, median age 30s. The most common presentation is a unilateral painful mass, which can ulcerate, fistulate and form abscesses. Management of IGM varies from expectant/topical steroids/NSAIDs to high dose steroids and DMARDs. A literature review shows that the usual starting dose of prednisolone is 40-60mg; the addition of methotrexate and azathioprine increases likelihood of successful treatment and allows for faster steroid tapering. Surgery for specific types of IGM can also be useful; for example, where there is duct granuloma, surgical excision can be curative. IGM can associate with arthritis, erythema nodosum, and episcleritis. One case series found that 14% of patients with IGM had concomitant arthritis. Key points: In our local experience we have had four patients with IGM, all previously treated with antibiotics or surgical methods but subsequently diagnosed with IGM and referred for immunosuppressive therapy. Being a rare condition, we feel it’s important for clinicians to be aware of IGM and its link with inflammatory arthritis. Disclosure S. Islam: None. P. Stonelake: None. H. John: None.

Associations of Early-Life and Adult Anthropometric Measures with the Expression of Stem Cell Markers CD44, CD24, and ALDH1A1 in Women with Benign Breast Biopsies.

According to the stem cell hypothesis, breast carcinogenesis may be related to the breast stem cell pool size. However, little is known about associations of breast cancer risk factors, such as anthropometric measures, with the expression of stem cell markers in noncancerous breast tissue. The analysis included 414 women with biopsy-confirmed benign breast disease in the Nurses' Health Study and Nurses' Health Study II. Birthweight, weight at age 18, current weight, and current height were reported via self-administered questionnaires. IHC staining of stem cell markers (CD44, CD24, and aldehyde dehydrogenase family 1 member A1) in histopathologically normal epithelial and stromal breast tissue was quantified using an automated computational image analysis system. Linear regression was used to examine the associations of early-life and adult anthropometric measures with log-transformed stem cell marker expression, adjusting for potential confounders. Birthweight [≥10.0 vs. <5.5 lbs: β (95% confidence interval) = 4.29 (1.02, 7.56); P trend = 0.001 in the stroma] and adult height [≥67.0 vs. <63.0 inch: 0.86 (0.14, 1.58); P trend = 0.02 in the epithelium and stroma combined] were positively associated with CD44 expression. Childhood body fatness was inversely associated (P trend = 0.03) whereas adult height was positively associated with CD24 expression in combined stroma and epithelium (P trend = 0.03). Our data suggest that anthropometric measures, such as birthweight, adult height, and childhood body fatness, may be associated with the stem cell expression among women with benign breast disease. Anthropometric measures, such as birthweight, height, and childhood body fatness, may have long-term impacts on stem cell population in the breast.

Multiplex Digital Spatial Profiling in Breast Cancer Research: State-of-the-Art Technologies and Applications across the Translational Science Spectrum.

While RNA sequencing and multi-omic approaches have significantly advanced cancer diagnosis and treatment, their limitation in preserving critical spatial information has been a notable drawback. This spatial context is essential for understanding cellular interactions and tissue dynamics. Multiplex digital spatial profiling (MDSP) technologies overcome this limitation by enabling the simultaneous analysis of transcriptome and proteome data within the intact spatial architecture of tissues. In breast cancer research, MDSP has emerged as a promising tool, revealing complex biological questions related to disease evolution, identifying biomarkers, and discovering drug targets. This review highlights the potential of MDSP to revolutionize clinical applications, ranging from risk assessment and diagnostics to prognostics, patient monitoring, and the customization of treatment strategies, including clinical trial guidance. We discuss the major MDSP techniques, their applications in breast cancer research, and their integration in clinical practice, addressing both their potential and current limitations. Emphasizing the strategic use of MDSP in risk stratification for women with benign breast disease, we also highlight its transformative potential in reshaping the landscape of breast cancer research and treatment.

A prospective cross-sectional observational comparative study: evaluating palpable breast lumps through comparative analysis of sonomammography breast imaging reporting and data system scores and fine needle aspiration cytology

This study protocol outlines a prospective cross-sectional observational investigation aimed at assessing the correlation between Sonomammography Breast Imaging Reporting and Data System (BI-RADS) scores and Fine-Needle Aspiration Cytology (FNAC) findings in diagnosing benign and malignant palpable breast lumps among patients admitted to AVBRH. The primary objective is to understand the diagnostic accuracy of FNAC and Sonomammography BI-RADS scoring in identifying breast lumps’ nature. Four specific objectives guide this study: 1) To determine the diagnostic accuracy of FNAC; 2) To evaluate the diagnostic precision of Sonomammography BI-RADS scores; 3) To compare the diagnostic outcomes of sonomammography and FNAC with HPE, and 4) To examine the age-wise distribution of benign and malignant palpable breast diseases. The study will be conducted at AVBRH, Sawangi (Meghe), Wardha, over three years, commencing in June 2022. The anticipated findings from this study will significantly contribute to refining the diagnostic strategies for palpable breast lumps, enhancing patient care and decision-making.

Involvement of FAM170B-AS1, hsa-miR-1202, and hsa-miR-146a-5p in breast cancer.

FAM170B-AS1 is usually expressed low in all organs except for testicular tissues. No study was performed to explore its role in breast cancer (BC). Contradictory results were reported about hsa-miR-1202 and hsa-miR-146a-5p in BC. The present study aimed to explore the involvement of FAM170B-AS1 in BC using bioinformatics predictive tools, followed by a practical validation besides exploring the impact of hsa-miR-1202 and hsa-miR-146a-5p in BC. This study enrolled 96 female patients with BC, 30 patients with benign breast diseases (BBD), and 25 control subjects. The expressions of circulating FAM170B-AS1, hsa-miR-1202, and hsa-miR-146a-5p were quantified using qRT-PCR. These ncRNAs' associations, predictive, and diagnostic roles in BC were statistically tested. The underlying miRNA/mRNA targets of FAM170B-AS1 in BC were bioinformatically predicted followed by confirmation based on the GEPIA and TCGA databases. The expression of FAM170B-AS1 was upregulated in sera of BC patients and hsa-miR-1202 was upregulated in sera of BBD and BC patients while that of hsa-miR-146a-5p was downregulated in BC. These FAM170B-AS1 was significantly associated with BC when compared to BBD. FAM170B-AS1 and hsa-miR-1202 were statistically associated with the BC's stage, grade, and LN metastasis. FAM170B-AS1 and hsa-miR-146a-5p gave the highest specificity and sensitivity for BC. KRAS and EGFR were predicted to be targeted by FAM170B-AS1 through interaction with hsa-miR-143-3p and hsa-miR-7-5p, respectively. Based on the TCGA database, cancer patients having mutations in FAM170B show good overall survival. The present study reported that for the first time, FAM170B-AS1 may be a potential risk factor, predictive, and diagnostic marker for BC. In addition, FAM170B-AS1 might be involved in BC by interacting with hsa-miR-143-3p/KRAS and hsa-miR-7-5p/EGFR through enhancement or repression that may present a new therapeutic option for BC.

Searching for a twinkling artefact in breast microcalcifications using B-flow ultrasonic non-doppler technology: a prospective study

INTRODUCTION: The rapid rejuvenation of breast cancer in recent years has made it necessary to look for non-X-ray methods of imaging microcalcifications in women not yet covered by mammography screening. Developing technologies to see these microcalcifications on ultrasound is a promising goal, potentially reducing the number of unnecessary biopsies.OBJECTIVE: Evaluation of factors influencing the ability of the B-flow technique to detect the presence of a flickering artifact in patients with microcalcifications detected by mammography.MATERIALS AND METHODS: A prospective study of 57 patients with microcalcifications detected on mammograms was conducted. The Nondoppler technology of B-flow ultrasound examination was used for targeted ultrasound with the search for these areas during ultrasound. The possibilities of visualization of the twinkle artifact were evaluated for various microcalcifications in size, distribution, their number, and for pathological processes of different nature. Images in B-flow mode were found to be positive when a bright local flash occurred against a dark background in B-mode. Verification of diagnostic findings was performed by ultrasound-guided core- biopsy from areas with a twinkle artifact (in the presence of an imaging area during ultrasound) or with stereotactic biopsy (in the absence of visualization of the suspicious area with ultrasound with B-flow).Statistics: The ANOVA test was used for triple pairwise comparisons between positive groups. The p&lt;0.05 value was considered statistically significant.RESULTS: According to the results of the study, out of 57 cases with microcalcifications, 34 (60%) recorded a twinkle artifact. The amount of microcalcifications with a visualized twinkle effect in B-flow was significantly higher in benign processes 34 (82.9%) out of 41, and less often in malignant 8 (50%) out of 16 (p=0.035). The effect of calcification morphology on the mammogram on their detection by ultrasound in the B-flow mode (p=0.035) was statistically significant. The most frequently manifested twinkle artifact were rounded, regular-shaped calcifications in 19 (95%) out of 20, which were found in granulomatous mastitis, fibrocystic changes, fibroadenomas, sclerosing adenosis, as well as polymorphic calcifications in 7 (63.6%) of 11. In the B-flow mode, it was not possible to obtain a twinkle artifact mainly with point small calcifications of 9 (69.2%) out of 13, and in all cases with linear or linear branching calcifications. In amorphous microcalcifications, the characteristics of background echogenicity and the presence or absence of a nodal component in the gray scale mode had a much greater influence on the appearance of the twinkle effect in the normal B-mode, whereas in the B-flow mode, these characteristics did not have a decisive effect on the appearance of a flickering artifact (p=0.8). The density of distribution of microcalcifications, the size of calcifications, the final pathomorphology, mammography characteristics, the background environment during ultrasound by the presence or absence of nodular formation and anechoic zones, did not affect the appearance of a bright flash in the B-flow mode.DISCUSSION: The density of calcification distribution was not decisive for the appearance of a shimmering artifact in the B-flow mode. The flicker artifact is recorded both in grouped microcalcifications and in single ones. B-flow can help to detect calcifications that are not visible in the usual B-flow mode due to the lack of pronounced tissue contrast. But this artifact cannot be used to differentiate between benign and malignant processes.CONCLUSION: The B-flow technique is promising for clarifying the twinkle artifact in the detected microcalcifications on the mammogram, which could potentially increase the incidence of ultrasound-guided trephine biopsies by detecting more areas with microcalcification accumulations.

Serum protein profiling reveals an inflammation signature as a predictor of early breast cancer survival

BackgroundBreast cancers exhibit considerable heterogeneity in their biology, immunology, and prognosis. Currently, no validated, serum protein-based tools are available to evaluate the prognosis of patients with early breast cancer.MethodsThe study population consisted of 521 early-stage breast cancer patients with a median follow-up of 8.9 years. Additionally, 61 patients with breast fibroadenoma or atypical ductal hyperplasia were included as controls. We used a proximity extension assay to measure the preoperative serum levels of 92 proteins associated with inflammatory and immune response processes. The invasive cancers were randomly split into discovery (n = 413) and validation (n = 108) cohorts for the statistical analyses.ResultsUsing LASSO regression, we identified a nine-protein signature (CCL8, CCL23, CCL28, CSCL10, S100A12, IL10, IL10RB, STAMPB2, and TNFβ) that predicted various survival endpoints more accurately than traditional prognostic factors. In the time-dependent analyses, the prognostic power of the model remained rather stable over time. We also developed and validated a 17-protein model with the potential to differentiate benign breast lesions from malignant lesions (Wilcoxon p < 2.2*10− 16; AUC 0.94).ConclusionsInflammation and immunity-related serum proteins have the potential to rise above the classical prognostic factors of early-stage breast cancer. They may also help to distinguish benign from malignant breast lesions.

RAPID ON-SITE EVALUATION AND MICRONUCLEUS SCORING IN BREAST LESIONS FINE NEEDLE ASPIRATION CYTOPATHOLOGY BY NEW IAC YOKOHAMA SYSTEM OF REPORTING

Objective: The objective of the study is to report all fine needle aspiration cytopathology (FNAC) cases of lump breast as per New Yokohama classification 2016, to highlight the importance of rapid onsite evaluation (ROSE) in breast cytology at the time of FNAC, and to assess the relationship between micronucleus (MN) scoring and various breast lesions in cytology. Methods: The study was conducted on 70 patients with breast lumps in the Department of Pathology in Government Medical College, Patiala, and comprised evaluation of ROSE and MN scoring in breast FNAC by the use of New Yokohama system of reporting. Results: On ROSE, 11 out of 70 patients were categorized under category I according to the Yokohama system of reporting and on the same sitting with repeat FNAC after performing the ROSE; nine cases were upgraded for cytological reporting and two remained in category I. According to as per the International Academy of Cytology (IAC) Yokohama system of reporting (2016) cases were categorized as category I – 2 (2.9%), category II – 46 (65.7%), category III – 7 (10.1%), category IV – 5 (7.1%), and category V – 10 (14.2%). In MN scoring, MN with the highest score of 14–17/1000 epithelial cells was in category V (malignant), and the lowest score was 0–1/1000 epithelial cells in category II (benign). Conclusion: ROSE is an easy, safe, and cost-effective method. IAC Yokohama system for reporting (2016) provides a comprehensive way of categorizing various breast lesions on FNAC with clinical correlation. MN score is a good biomarker in differentiating benign, atypical, and malignant breast lesions.

Abstract LB328: Measurement of ER and Ki67 coexpression in cytokeratin positive epithelial cells in benign breast disease (BBD) percutaneous biopsies

Abstract Over 1 million women in the U.S. undergo radiologically guided percutaneous biopsies annually. Of these, 75% reveal benign breast disease (BBD), typically surrounded by normal breast lobules, which are proposed to reflect the biology of the breast parenchyma diffusely. With aging, normal lobules undergo involution, and delays in this process are linked to increased breast cancer (BC) risk. Because BBD patients are at increased risk of developing BC, and pathologic classification only stratifies risks among groups of patients, novel methods for individual BC risk prediction are needed to guide precision management. Prior literature has shown that most proliferating epithelial cells in normal lobules do not express estrogen receptor (i.e., ER-/Ki67+), whereas co-expression is frequent in BC precursors and in BCs (i.e., ER+/Ki67+). Thus we developed and applied a novel quantitative dual ER/Ki67 stain to BBD biopsies from the well-characterized contemporary Mayo Clinic BBD cohort (2002-2013) to assess differences by levels of lobular involution and tissue type (BBD vs. normal), as well as associations with BC risk. We identified 100 BBD biopsies preceding BC (cases) and 100 biopsies from cancer-free women (controls), frequency-matched on age and length of follow-up. A multiplex immunofluorescence assay was developed to detect ER/Ki67 coexpression in cytokeratin positive epithelial cells in normal lobules or BBD lesions in FFPE tissue sections. An inverse normal transformation was applied to cell counts and proportions and associations were assessed for levels of lobular involution, tissue type (normal lobule vs. BBD), and case status using t-tests and linear regression models. The final data set included 1152 normal lobules or BBD lesions in biopsies of 177 subjects (88 cases and 89 controls). In univariate analyses, normal lobules showing delayed age related involution included higher counts of ER-/Ki67+, ER+/Ki67- and ER+/Ki67+ cells, and the lowest proportion of ER+/Ki67- cells (p&amp;lt;0.03; all). In separate analyses comparing presence vs. absence, detection of any ER+/Ki67+ cells was suggestively more common in non-involuted vs. involuted lobules. Compared with normal lobules, BBD showed higher counts and proportions of ER+/Ki67- and ER+/Ki67+ cells and lower values for ER-/Ki67+ (all p&amp;lt;0.001). In these preliminary results, marker expression in normal lobules and in BBD did not differ by case-control status. Preliminarily, we find that ER+/Ki67+ cells are increased in normal lobules with delayed age-related involution and in BBD lesions, consistent with their associations with increased BC risk. These findings are consistent with the risk reducing effects of endocrine chemoprevention and may serve as useful biomarkers in prevention therapy. In this analysis, limited to only a few lobules and BBD lesions per biopsy, ER+/Ki67+ cell counts and proportions did not vary significantly by case-control status. Citation Format: Melody L. Stallings-Mann, Robert A. Vierkant, Stacey J. Winham, Bryan McCauley, Matt R. Jensen, Sophie Noel, Laura Pacheco-Spann, Celine M. Vachon, Amy C. Degnim, Derek C. Radisky, Mark E. Sherman. Measurement of ER and Ki67 coexpression in cytokeratin positive epithelial cells in benign breast disease (BBD) percutaneous biopsies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB328.

Role of P63 in Benign and Malignant Lesions of Breast

Introduction: Breast carcinoma is leading cause of cancer death in women. Breast lesions constitute heterogeneous group of diseases with wide variety of etiologies ranging from inflammatory-benign- malignant lesions. There are several reported markers for immunohistochemical detection of myoepithelial cells. Smooth muscle specific proteins, such as smooth muscle actin, smooth muscle myosin heavy chain, calponin and h-caldesmon are used to highlight myoepithelium. p63 antibody is myoepithelial cell marker that selectively stains nuclei. It is negative in stromal, myofibroblastic and adipocytic cells.Aims and Objectives: The aim of this study was to establish role of p63 expression in distinguishing benign breast lesions, premalignant lesions and malignant tumors of breast.Materials and Methods: 30 cases were selected from core biopsy, lumpectomy and mastectomy specimens ofbreast received at department of Pathology, Chalmeda Anand Rao Institute of Medical Sciences during the period of January 2021 to December 2021 and were studied prospectively. All specimens were processed according to CAP protocol and reported . Immunohistochemistry was performed to determine p63 expression in those specimens. p63 expression was evaluated as continuous positive/discontinuous positive/negative.Ethical Approval: This study was reviewed and approved by institute ethics committee, CAIMS, Karimnagar.Results: Among total 30 cases, 18 cases (60%) were benign lesions and all were positive for p63 expression. 3 cases (9.99%) were premalignant and were least positive for p63 expression. All malignant cases 9 cases (29.99%) were negative for p63 expression.Conclusion: The Positive correlation was seen between histomorphological features and p63 scoring in all the lesions, So p63 is good Immunohistochemical marker for evaluating breast lesions.

Abstract CT273: Unique multicenter trial design to investigate ARTIDIS nanomechanical generated measurements for early breast lesions (ANGEL)

Abstract Background: The conventional diagnosis of breast cancer followed by molecular subtyping for therapy selection is a lengthy process and associated with patient and clinician anxiety. Efficient, bedside approaches are desirable to drive fast diagnosis and early planning of clinical management. This clinical trial (NCT06085833) tackles the novel concept of automated and reliable tissue diagnostics (ARTIDIS) based on the measurement of nanomechanical signature of fresh clinical biopsies from patients with breast lesions. This study is based on our previous single center clinical study (Basel NANO) where tissue nanomechanical signature can: 1) reliably distinguish benign and malignant breast lesions, 2) predict the presence of metastatic disease based on imminent aggressiveness rating and 3) predict the response to neoadjuvant chemotherapy from baseline biopsies. The current study aims to prospectively validate the efficacy of ARTIDIS system as an aid to diagnosis, assessment of breast cancer prognosis and prediction of therapy response, particularly in the context of neoadjuvant therapy (Natira). Methods: A cohort of 2,705 patients with breast lesions, who are clinically indicated for core needle biopsy (CNB) are being enrolled, from diverse ethnic and socioeconomic backgrounds. Single fresh CNB will be transferred into tissue preservation buffer and measured by ARTIDIS device within 1-3 hours to collect thousands of nanomechanical measurements. Undisrupted CNB will be fixed and returned to standard diagnostic workflow, remaining unaltered for downstream assessments. Nanomechanical measurement data are prospectively analyzed in a blind manner before the collection of histopathology and follow up clinical data. The primary endpoint of the study is to validate ARTIDIS potential in diagnosis of breast lesions (benign vs malignant), secondary endpoints address whether nanomechanical signature can differentiate subtypes of breast cancer, associate with clinical outcome and guide treatment decisions in NAT setting. Follow up data will be collected regularly up to 10 years after patient enrollment. Results: The multicenter ANGEL study is a prospective validation of the NANO (University Hospital Basel, Switzerland) - first ever performed clinical study which investigated the diagnostic, prognostic, and predictive value of tissue nanomechanical signature in patients with breast cancer. The ANGEL study takes advantage of ARTIDIS non-disruptive technology and is fully integrated into routine clinical workflow. The study started enrollment in 11/02/2023, and is expected to complete by 2025. The seamless integration of ARTIDIS within the standard clinical settings, its automatization, bedside implementation, and the lack of additional burden to patients enables effective reach of a diverse population through local hospitals and communities. Conclusion: This study presents the first of its kind as a direct prospective, multicenter- based translation of the tissue nanomechanical signature into clinical settings. This unique trial design allows the prospective development of ARTIDIS as an aid to diagnosis, prognosis and therapy optimization in patients with breast cancer. The innovative trial design of this study may also further the development of strategies to increase reach to underrepresented communities. Citation Format: Karla A. Sepulveda, Ashley Roark, Sagar Dhamne, Chandandeep Nagi, Ivan Marin, Iqbal Tabish, Sara Nizzero, Mariam Gachechiladze, Tobias Appenzeller, Philipp Oertle, Marko Loparic, Susan G. Hilsenbeck, Marija Plodinec, Alastair Thompson. Unique multicenter trial design to investigate ARTIDIS nanomechanical generated measurements for early breast lesions (ANGEL) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT273.

Osseous Metaplasia of the Breast: A Series of Rare Cases

Osseous metaplasia is a reversible condition in which non-osseous soft tissue is replaced by bone. This is most commonly seen in musculoskeletal system and central nervous system. Osseous metaplasia in breast is a rare event. In this case series, we have reported 4 cases of osseous metaplasia of breast. 1 case of Osseous metaplasia in benign phyllodes tumor, 1 case of Osseous metaplasia in fibroadenoma, 1 case of Osseous metaplasia in invasive breast carcinoma and 1 extremely rare case of Osseous metaplasia of breast presenting as breast lump without any other associated pathology. Since osseous metaplasia of breast is a rare phenomenon, this entity specially in association with benign breast disease conditions pose a great challenge for accurate diagnosis.

Development and validation of a model and nomogram for breast cancer diagnosis based on quantitative analysis of serum disease-specific haptoglobin N-glycosylation

BackgroundA better diagnostic marker is in need to distinguish breast cancer from suspicious breast lesions. The abnormal glycosylation of haptoglobin has been documented to assist cancer diagnosis. This study aims to evaluate disease-specific haptoglobin (DSHp)-β N-glycosylation as a potential biomarker for breast cancer diagnosis.MethodsDSHp-β chains of 497 patients with suspicious breast lesions who underwent breast surgery were separated from serum immunoinflammatory-related protein complexes. DSHp-β N-glycosylation was quantified by mass spectrometric analysis. After missing data imputation and propensity score matching, patients were randomly assigned to the training set (n = 269) and validation set (n = 113). Logistic regression analysis was employed in model and nomogram construction. The diagnostic performance was analyzed with receiver operating characteristic and calibration curves.Results95 N-glycopeptides at glycosylation sites N207/N211, N241, and N184 were identified in 235 patients with benign breast diseases and 262 patients with breast cancer. DSHp-β N-tetrafucosyl and hexafucosyl were significantly increased in breast cancer compared with benign diseases (p < 0.001 and p = 0.001, respectively). The new diagnostic model and nomogram included GN2F2, G6N3F6, GN2FS at N184, G-N&G2S2, G2&G3NFS, G2N3F, GN3 at N207/N211, CEA, CA153, and could reliably distinguish breast cancer from benign diseases. For the training set, validation set, and training and validation sets, the area under the curves (AUCs) were 0.80 (95% CI: 0.75–0.86, specificity: 87%, sensitivity: 62%), 0.77 (95% CI:0.69–0.86, specificity: 75%, sensitivity: 69%), and 0.80 (95% CI:0.76–0.84, specificity: 77%, sensitivity: 68%), respectively. CEA, CA153, and their combination yielded AUCs of 0.62 (95% CI: 0.56–0.67, specificity: 29%, sensitivity: 90%), 0.65 (95% CI: 0.60–0.71, specificity: 74%, sensitivity: 51%), and 0.67 (95% CI: 0.62–0.73, specificity: 60%, sensitivity: 68%), respectively.ConclusionsThe combination of DSHp-β N-glycopeptides, CEA, and CA153 might be a better serologic marker to differentiate between breast cancer and benign breast diseases. The dysregulated N-glycosylation of serum DSHp-β could provide insights into breast tumorigenesis.

Study on the differential diagnosis of benign and malignant breast lesions using a deep learning model based on multimodal images.

To establish a multimodal model for distinguishing benign and malignant breast lesions. Clinical data, mammography, and MRI images (including T2WI, diffusion-weighted images (DWI), apparent diffusion coefficient (ADC), and DCE-MRI images) of 132 benign and breast cancer patients were analyzed retrospectively. The region of interest (ROI) in each image was marked and segmented using MATLAB software. The mammography, T2WI, DWI, ADC, and DCE-MRI models based on the ResNet34 network were trained. Using an integrated learning method, the five models were used as a basic model, and voting methods were used to construct a multimodal model. The dataset was divided into a training set and a prediction set. The accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of the model were calculated. The diagnostic efficacy of each model was analyzed using a receiver operating characteristic curve (ROC) and an area under the curve (AUC). The diagnostic value was determined by the DeLong test with statistically significant differences set at P < 0.05. We evaluated the ability of the model to classify benign and malignant tumors using the test set. The AUC values of the multimodal model, mammography model, T2WI model, DWI model, ADC model and DCE-MRI model were 0.943, 0.645, 0.595, 0.905, 0.900, and 0.865, respectively. The diagnostic ability of the multimodal model was significantly higher compared with that of the mammography and T2WI models. However, compared with the DWI, ADC, and DCE-MRI models, there was no significant difference in the diagnostic ability of these models. Our deep learning model based on multimodal image training has practical value for the diagnosis of benign and malignant breast lesions.

A Study Comparing Centchroman and Evening Primrose Oil in the Treatment of Benign Breast Disease

ABSTRACT Introduction: Females seeking medical attention at breast clinics commonly present with nonmalignant breast conditions, including nonspecific breast pain or distinct masses like fibroadenomas. Due to its variability, it may be difficult to quantify breast pain. The purpose of the research was to compare centchroman and evening primrose oil to identify a cost-effective, secure, and efficient treatment for benign breast disease. Material and Methods: In this prospective hospital-based observational study, 100 breast diseases with or without lumpiness for 1 year were included and divided into two groups with 50 cases each, Group-A (Centchroman) and Group-B (Evening primrose oil). Results: Centchroman exhibited a significantly greater treatment response for alleviating pain-free mastalgia compared to evening primrose oil. Additionally, centchroman showed an excellent response (P &lt; .05). Among participants with mastalgia, centchroman significantly reduced the number of mastalgia patients with tender nodularity post-treatment (P = .035) than evening primrose oil. On the basis of fibroadenoma, partial and complete response was significantly seen in higher number of cases in the centchroman group (P = .007). Conclusion: Centchroman therapy demonstrates that the treatment for benign breast disease is safe, effective, and economical.

396 Unraveling the Immunological Basis of Lobular Involution in Breast Cancer Development

OBJECTIVES/GOALS: Reveal common immune mechanisms in dysregulated age-related lobular involution (ARLI) and post-partum lobular involution (PPLI) to understand their link to increased breast cancer risk, challenging the traditional view of their distinctiveness. Ultimately, to improve breast cancer risk assessment and personalized prevention METHODS/STUDY POPULATION: The Mayo Clinic Benign Breast Disease (BBD) cohort comprises of ~20,000 women with benign biopsies, including ~1000 women with sequential benign biopsies. Lobular involution (LI) status was assessed by selecting perimenopausal women, ages 45-55, with sequential biopsies, comparing acini number and lobule size between initial and subsequent biopsies. NanoString IO360/ BC360 RNA profiling identified differentially expressed genes associated with dysregulated LI. Using multiplex immunofluorescence (mIF), I'll analyze and spatially map immune biomarkers related to dysregulated ARLI and PPLI in BBD tissue from perimenopausal women who did or did not go on to develop breast cancer, assessing the commonality of ARLI and PPLI markers and exploring their potential as risk biomarkers for breast cancer. RESULTS/ANTICIPATED RESULTS: Preliminary findings link patients who display dysregulated ARLI with an increased breast cancer risk and identify vital PPLI biomarkers in perimenopausal women. I expect the biopsies of women who developed post-menopausal breast cancer (PMBC) and post-partum breast cancer (PPBC) to exhibit elevated levels of dysregulated ARLI immune biomarkers and PPLI biomarkers. Spatially mapping these markers promises to provide a more comprehensive understanding of their interactions, potentially revealing common immunological pathways. These findings could transform our current paradigm of ARLI and PPLI as distinct processes and demonstrate their interconnection in shaping breast cancer risk. DISCUSSION/SIGNIFICANCE: PMBC and PPBC dominate majority of breast cancer cases. Both involve activation of the understudied process of lobular involution, which has been shown to have pro-tumorigenic traits. Elucidating these mechanisms will aid more efficient risk stratification and personalized prevention to reduce incidence and mortality of breast cancer.