Early-stage Breast Tumors Research Articles

Abstract PO4-22-02: pSmad2 as a marker of radiotherapy benefit in breast cancer- results from the randomized SweBCG91RT trial

Abstract Background: The TGF-beta pathway can have both pro- and antitumoral effects in breast cancer. Experimental studies further suggest that TGF-beta can mediate radioresistance, either through direct effects on tumor cells or indirectly by its immunomodulatory properties. However, the effect of TGF-beta signaling is highly depending on tumor stage, mutational status and microenvironmental cell composition. In order to explore TGF-beta signaling as a potential biomarker or therapeutical target it is therefore important to perform analyses in well-defined study populations. The purpose of this study was to study the effect of active TGF beta signaling in tumor cells, on the risk of ipsilateral breast tumor recurrence (IBTR) and benefit from radiotherapy (RT) in specifically early-stage breast cancer using nuclear, phosphorylated Smad2 (pSmad2) as a marker. Methods: The SweBCG91RT cohort included 1179 patients with stage I-IIA tumors randomized to breast-conserving surgery (BCS) with or without postoperative RT. Nuclear pSmad2 expression was assessed as for 987 available tumors on TMAs as the mean proportion of tumor cells with positive nuclear staining. Based on the distributions, tumors were categorized as having high (≥80%, n=344), medium (21-79%, n=428) or low (≤20%, n=161) pSmad2 staining. The primary endpoint was time to ipsilateral breast tumor recurrence as the first event within 10 years. Results: pSmad2Low tumors were more likely to be of grade III (p< 0.001) and tended to be larger than pSmad2Medium (p=0.002, median 15 vs 12 mm) and pSmad2High (p< 0.001, median 15 vs 12 mm) tumors. No associations between pSmad2 and subtype or age were found. Numerically, tumors with pSmad2low (HR 1.55, CI 95% 0.80-3.02, p=0.19) or pSmad2Medium (HR 2.53, CI 95% 1.52-4.22, p< 0.001) had a higher risk of IBTR than pSmad2high tumors (reference). These findings remained significant after adjustment for age, histological grade, and RT. The benefit from RT was similar for pSmad2High (HR0.41, CI 95% 0.15-1.07, p=0.068), pSmad2Medium (HR 0.36, CI 95% 0.21-0.63, p< 0.001), and pSmad2Low (HR 0.51, CI 95% 0.18-1.46, p=0.21) tumors (pinteraction=0.86). Conclusions: Early-stage breast tumors with high nuclear pSmad2 staining may be at a reduced risk of an IBTR. However, the relative benefit from RT does not seem to differ from tumors with medium or low pSmad2 staining. These results need to be confirmed in additional studies, but they provide initial evidence that tumoral TGF-beta signaling has rather tumor restraining functions in early-stage breast cancer. Next analytical steps should include the tumor mutation status (particularly for TGF-beta signaling components TGFBR2, SMAD4 and TP53), as well as the TGF-beta signaling status within the tumor microenvironment. Citation Format: Per Karlsson, Viktoria Thurfjell, Anikó Kovács, Fredrika Killander, Emma Niméus, Erik Holmberg, Carina Strell, Axel Stenmark Tullberg. pSmad2 as a marker of radiotherapy benefit in breast cancer- results from the randomized SweBCG91RT 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 PO4-22-02.

Abstract PO2-13-08: Cell-free orphan noncoding RNAs and AI enable early detection of invasive breast cancer and ductal carcinoma in-situ

Abstract Background Earlier detection of breast cancer through mammography screening has reduced disease-specific mortality; however, confounding issues such as technical challenges, breast density, and tumor size can result in false negatives and ultimately later stage diagnosis. Next generation liquid biopsy has the potential to complement mammography and enable earlier detection for more women. We have previously demonstrated high sensitivity and specificity for early detection of invasive breast cancer (IBC) by utilizing a novel category of cancer-associated small RNAs, termed orphan noncoding RNAs (oncRNAs), through a liquid biopsy platform. Here, we further improve the ability to detect breast cancer in a larger, multi-source cohort through an AI-driven approach and demonstrate potential for detection of ductal carcinoma in-situ (DCIS). Methods We utilized The Cancer Genome Atlas (TCGA) small RNA-seq database to discover a library of 20,538 oncRNAs, through a female-specific analysis, that were significantly enriched among 1,103 breast tumors compared to 349 normal tissue samples spanning multiple tissue sites. The diagnostic performance of these oncRNAs were assessed in an independent cohort of archived serum samples from 663 female individuals, sourced from Indivumed (Hamburg, Germany), Proteogenex (Inglewood, CA), and MT Group (Los Angeles, CA), including 279 breast cancer patients of various stages (221 IBC and 58 DCIS; mean age: 57.0 ± 13.8 years; ever-smoker: 25.8%) and 304 age-matched controls (mean age: 58.5 ± 13.9 years; ever-smoker: 23.4%) without breast cancer. All samples were collected between 2010–2022 at time of diagnosis for breast cancer patients. We sequenced the small RNA content of these samples at an average depth of 25.28 ± 9.37 million 50-bp single-end reads. We detected 18,025 (87.8%) unique breast cancer-specific oncRNA species within at least one sample from the study cohort. We then trained a generative AI model using 5-fold cross-validation to predict cancer status for all samples. Results Our oncRNA-based model achieved an overall AUC of 0.95 (95% CI, 0.93–0.97) for prediction of IBC versus cancer-free controls with a sensitivity of 0.87 (0.82–0.91) at 90% specificity. We observed high sensitivities, also at 90% specificity, across all tumor stages and tumor sizes (Table 1). Sensitivities for the earliest stage and smallest tumor size were 0.87 (0.78–0.93) and 0.81 (0.61–0.93) for Stage I (n=83) and T1a–b ( >1mm to ≤10mm; n=26), respectively. Additionally, in a small single-source cohort, we also saw high model accuracy and sensitivity for DCIS, which we aim to confirm in additional cohorts. While our overall cancer cohort primarily consisted of individuals with luminal breast cancer, our model had high sensitivities across all breast cancer subtypes at 0.90 (0.84–0.94), 0.73 (0.59–0.85), and 0.86 (0.42–1.0) for luminal (n=181), HER2 positive (n=49), and triple negative (n=7), respectively. Conclusions We further demonstrate the potential utility of oncRNAs as a blood-based biomarker using an AI algorithm for sensitive and accurate early detection of breast cancer in a large cohort. Additionally, we have shown that this oncRNA-based assay performs well in detecting small, early-stage invasive breast tumors, with potential to detect precursors of breast cancer. Table 1: Model sensitivity in breast cancer by tumor stage and size For each tumor stage and size, as defined by the AJCC 7th Edition breast cancer staging system, sensitivity and 95% Pearson-Clopper confidence intervals (CI) are reported at 90% specificity for the number of samples (N). Citation Format: Noura Tbeileh, Taylor Cavazos, Mehran Karimzadeh, Jeffrey Wang, Alice Huang, Dung Ngoc Lam, Seda Kilinc, Jieyang Wang, Xuan Zhao, Andy Pohl, Helen Li, Lisa Fish, Kimberly Chau, Marra Francis, Lee Schwartzberg, Patrick Arensdorf, Hani Goodarzi, Fereydoun Hormozdiari, Babak Alipanahi. Cell-free orphan noncoding RNAs and AI enable early detection of invasive breast cancer and ductal carcinoma in-situ [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-13-08.

Perturbation and stability of PAM50 subtyping in population-based primary invasive breast cancer

PAM50 gene expression subtypes represent a cornerstone in the molecular classification of breast cancer and are included in risk prediction models to guide therapy. We aimed to illustrate the impact of included genes and biological processes on subtyping while considering a tumor’s underlying clinical subgroup defined by ER, PR, and HER2 status. To do this we used a population-representative and clinically annotated early-stage breast tumor cohort of 6233 samples profiled by RNA sequencing and applied a perturbation strategy of excluding co-expressed genes (gene sets). We demonstrate how PAM50 nearest-centroid classification depends on biological processes present across, but also within, ER/PR/HER2 subgroups and PAM50 subtypes themselves. Our analysis highlights several key aspects of PAM50 classification. Firstly, we demonstrate the tight connection between a tumor’s nearest and second-nearest PAM50 centroid. Additionally, we show that the second-best subtype is associated with overall survival in ER-positive, HER2-negative, and node-negative disease. We also note that ERBB2 expression has little impact on PAM50 classification in HER2-positive disease regardless of ER status and that the Basal subtype is highly stable in contrast to the Normal subtype. Improved consciousness of the commonly used PAM50 subtyping scheme will aid in our understanding and interpretation of breast tumors that have seemingly conflicting PAM50 classification when compared to clinical biomarkers. Finally, our study adds further support in challenging the common misconception that PAM50 subtypes are distinct classes by illustrating that PAM50 subtypes in tumors represent a continuum with prognostic implications.

Reduced racial disparity in receipt of optimal locoregional treatment for women with early-stage breast cancer.

Racial disparities in breast cancer treatment contribute to Black women having the worst breast cancer survival rates in the U.S. We investigated whether differences in receipt of optimal locoregional treatment (OLT), defined as receipt of mastectomy, breast-conserving surgery, or no surgery when contraindicated, existed between Black and White women with early-stage breast cancer from 2008-2018. In this retrospective cohort study, data from the Surveillance, Epidemiology, and End Results (SEER) Program Incidence Database was utilized to identify tumor cases from Black and White women aged 20-64 years old with stage I-II breast cancer. Logistic regression analyses were used to evaluate the associations between race and receipt of OLT as well as potential effect modification by tumor characteristics, and year of diagnosis. Among 177,234 women diagnosed with early-stage breast tumors, disparities in OLT between Black and White women were present from 2008-2010 (2008: 82.1% Black vs. 85.7% White, p<0.001; 2009: 82.1% Black vs. 85.8% White, p<0.001; 2010: 82.2% Black vs. 87.2% White, p<0.001). This disparity was eliminated between 2010-2011 (86.3% Black vs. 87.5% White, p = 0.15), and did not reoccur during the remainder of the study period. From 2010-2011, more Black women received radiation therapy following breast-conserving surgery (43.4% to 48.9%; p = 0.001), which accounted for an overall increased receipt of OLT. Increased receipt of radiation therapy with breast-conserving surgery appeared to drive a substantial increase in OLT for Black women from 2010-2011 that lasted throughout the study period. Further research on the underlying mechanisms that reduced this disparity is warranted.

Abstract PD18-04: Prognostic implications of PIK3CA mutation by hormone receptor status and intrinsic subtype in early stage HER2-positive breast cancer: a correlative analysis from CALGB 40601

Abstract Background PIK3CA mutations have been described in 20-25% of early-stage HER2-positive breast tumors [1], and are associated with reduced pathologic complete response (pCR) rate after chemotherapy and anti-HER2 agents [2]. However, the independence of this finding and association with long-term outcomes within HER2+ patients is still largely unknown. Here, we studied the prognostic implications of PIK3CA mutations by hormone receptor (HR) status and intrinsic subtype in patients with early stage HER2+ breast cancer enrolled in CALGB 40601. Method In CALGB 40601, gene expression profiling by RNA sequencing (RNAseq) with PAM50-determined intrinsic subtype and PIK3CA mutations from whole exome sequencing (WES) were obtained from 184/305 (60%) pretreatment core biopsies. We examined the association of PIK3CA mutations with pCR and event free survival (EFS) by HR status and intrinsic subtype using logistic and Cox regression analyses. Results PIK3CA mutations were found in 32 patients (32/184, 17%). The most frequent mutation was H1047R (12/32,38%), followed by E545K (7/32,22%) and E542K (5/32,16%). PIK3CA mutations were present in 20% and 15% of HR-positive and HR-negative BC subpopulations, respectively. Within Luminal-B, Luminal-A and HER2-Enriched breast tumors, PIK3CA mutations occurred in 36%, 10% and 19% respectively. In the overall population there was lower rate of pCR in mutated-PIK3CA patients than wild-type (34% vs 49%). Using only the subset of patients treated with neoadjuvant trastuzumab-based therapy as standard of care (excluding the lapatinib plus paclitaxel arm), we found a statistically significant lower pCR rate among PIK3CA-mutated tumors using logistic regression model (30% vs 54%, OR=0.30, p=0.045). At a median follow-up of 9.1 years, the presence of PIK3CA mutation was significantly associated with worse EFS in the overall study population (HR 2.58, 95% CI 1.24- 5.35, p=0.011). In a multivariable model including pCR status, HR status and intrinsic subtype (HER2-E vs. not), PIK3CA mutation was independently and significantly associated with worse EFS (HR 2.18, 95% CI 1.04- 4.56, p=0.039). The negative impact of PIK3CA mutation on EFS was statistically significant only in patients with HR-positive (HR 3.6, 95% CI 1.45-8.96, p=0.06) and luminal breast tumors (HR 4.84, 95% CI 1.08-21.7, p=0.039), but not in HR-negative and non-luminal subtypes. Conclusion In our study, the presence of PIK3CA mutation was significantly associated with lower pCR rates in patients treated with chemotherapy plus trastuzumab. Moreover, in uni- and multivariable Cox models, PIK3CA mutations were associated with worse long-term survival, which appeared to be driven by HR-positive and luminal HER2-positive breast tumors.

Identifying distinctive tissue and fecal microbial signatures and the tumor-promoting effects of deoxycholic acid on breast cancer.

A growing body of evidence indicates that the dysbiosis of both mammary and intestinal microbiota is associated with the initiation and progression of breast tumors. However, the microbial characteristics of patients with breast tumors vary widely across studies, and replicable biomarkers for early-stage breast tumor diagnosis remain elusive. We demonstrate a machine learning-based method for the analysis of breast tissue and gut microbial differences among patients with benign breast disease, patients with breast cancer (BC), and healthy individuals using 16S rRNA sequence data retrieved from eight studies. QIIME 2.0 and R software (version 3.6.1) were used for consistent processing. A naive Bayes classifier was trained on the RDP v16 reference database to assign taxonomy using the Vsearch software. After re-analyzing with a total of 768 breast tissue samples and 1,311 fecal samples, we confirmed that Halomonas and Shewanella were the most representative genera of BC tissue. Bacteroides are frequently and significantly enriched in the intestines of patients with breast tumor. The areas under the curve (AUCs) of random forest models were 74.27% and 68.08% for breast carcinoma tissues and stool samples, respectively. The model was validated for effectiveness via cohort-to-cohort transfer (average AUC =0.65) and leave-one-cohort-out (average AUC = 0.66). The same BC-associated biomarker Clostridium_XlVa exists in the tissues and the gut. The results of the in-vitro experiments showed that the Clostridium-specific-related metabolite deoxycholic acid (DCA) promotes the proliferation of HER2-positive BC cells and stimulates G0/G1 phase cells to enter the S phase, which may be related to the activation of peptide-O-fucosyltransferase activity functions and the neuroactive ligand-receptor interaction pathway. The results of this study will improve our understanding of the microbial profile of breast tumors. Changes in the microbial population may be present in both the tissues and the gut of patients with BC, and specific markers could aid in the early diagnosis of BC. The findings from in-vitro experiments confirmed that Clostridium-specific metabolite DCA promotes the proliferation of BC cells. We propose the use of stool-based biomarkers in clinical application as a non-invasive and convenient diagnostic method.

Low-molecular-weight cyclin E deregulates DNA replication and damage repair to promote genomic instability in breast cancer.

Low-molecular-weight cyclin E (LMW-E) is an N-terminus deleted (40 amino acid) form of cyclin E detected in breast cancer, but not in normal cells or tissues. LMW-E overexpression predicts poor survival in breast cancer patients independent of tumor proliferation rate, but the oncogenic mechanism of LMW-E and its unique function(s) independent of full-length cyclin E (FL-cycE) remain unclear. In the current study, we found LMW-E was associated with genomic instability in early-stage breast tumors (n = 725) and promoted genomic instability in human mammary epithelial cells (hMECs). Mechanistically, FL-cycE overexpression inhibited the proliferation of hMECs by replication stress and DNA damage accumulation, but LMW-E facilitated replication stress tolerance by upregulating DNA replication and damage repair. Specifically, LMW-E interacted with chromatin and upregulated the loading of minichromosome maintenance complex proteins (MCMs) in a CDC6 dependent manner and promoted DNA repair in a RAD51- and C17orf53-dependent manner. Targeting the ATR-CHK1-RAD51 pathway with ATR inhibitor (ceralasertib), CHK1 inhibitor (rabusertib), or RAD51 inhibitor (B02) significantly decreased the viability of LMW-E-overexpressing hMECs and breast cancer cells. Collectively, our findings delineate a novel role for LMW-E in tumorigenesis mediated by replication stress tolerance and genomic instability, providing novel therapeutic strategies for LMW-E-overexpressing breast cancers.

Low-Cost Antenna-Array-Based Metamaterials for Non-Invasive Early-Stage Breast Tumor Detection in the Human Body.

Microstrip patch antennas have been used in many applications since their appearance. Despite their great promise, their narrow bandwidth and the loss at high-frequency bands have limited their usage in medical applications. This work proposes a developed low-cost microstrip patch antenna suitable for microwave imaging (MWI) applications within the wideband frequency range. The proposed antenna is loaded with an artificial magnetic conductor (AMC) to improve the antenna performance. The simulated results obtained using computer simulation technology (CST) indicate that the presence of the AMC has improved the frequency selectivity of the antenna at 8.6 GHz with a peak realized gain of 9.90443 dBi and 10.61 dBi for simulated and measured results, respectively. The proposed microstrip antenna has been fabricated to validate the simulated results, and its performance is tested experimentally. Additionally, the fidelity factor of face-to-face (FtF) and side-by-side (SbS) scenarios have been presented. The breast phantom models with a tumor and the antenna operating as a transceiver have been numerically simulated for the application of cancer tumor cell detection. The work will have a significant impact on the design of electromagnetic biosensors.

A compact ultrawideband slotted patch antenna for early stage breast tumor detection applications

AbstractThis paper presents a compact ultrawideband slotted patch antenna for early stage tumor detection applications using microwave breast imaging systems. In order to reduce any reflections between the antenna and breast, the antenna is designed and optimized inside a coupling medium with electrical properties similar to that of normal breast tissue as a coupling medium. The wideband performance of the antenna is obtained by adding a rectangular slit and L-shaped sleeves on the ground plane. While the added rectangular slit mainly improves the matching, the L-shaped sleeves improve both the matching and the bandwidth. The antenna operates from 0.9 to 9.6 GHz with a fractional bandwidth of ~165$\%$, providing a good penetration and resolution. In addition, the proposed antenna has a very compact size of 16 × 16 mm2. Furthermore, an antenna array consisting of 31 antennas is formed around a realistic breast phantom in the simulation environment to demonstrate the imaging of a spherical tumor with a diameter of 5 mm inside the breast. The results confirm that the proposed antenna can be used for early stage breast tumor detection purposes.

Abstract 5030: Intra-patient tumor evolution analysis in dogs with many mammary tumors identifies signatures of tumor aggression in early-stage breast cancers

Abstract Despite tremendous improvements in breast cancer detection, it remains unclear which early-stage tumors will later become aggressive. Dogs provide a unique opportunity to unravel early-stage tumor evolution, as dogs frequently develop multiple naturally occurring mammary tumors. To identify which early-stage human breast tumors will later become malignant, we analyzed mutational profiles of mammary tumors from two canine cohorts to identify patterns of tumor evolution. We compared our dog tumor signatures with human breast cancers. We found that dogs and humans share many known cancer driver mutations. Additionally, three of the five dogs with at least five mammary tumors had a single driver mutation present in most of their tumors. Despite sharing patient-level exposures and environment, passenger mutations significantly differ between these tumors, suggesting independent primaries initiated by a catalyzing factor causing the same driver mutation in each tumor. However, tumors from the same dog with the same driver gene mutations rarely exhibit the same expression-based breast cancer subtypes. Our analysis highlights the mutational similarities of dog and human breast cancers and provides insight into their development. Citation Format: Kiley Graim. Intra-patient tumor evolution analysis in dogs with many mammary tumors identifies signatures of tumor aggression in early-stage breast cancers [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 5030.

Abstract P3-20-04: High-resolution 18F-FDG PET-CT for peri-operative margin assessment in early-stage breast cancer: Results of an ongoing clinical study

Abstract Introduction Successful breast-conserving surgery (BCS) entails excision of the breast tumor with an adequate amount of surrounding healthy breast tissue, such that negative resection margins are obtained. Despite efforts to obtain tumor-free margins, approximately 20% of the breast cancer patients still require reoperation. In this study, we investigate high-resolution 18F-fluoro-deoxyglucose (FDG) positron emission tomography (PET) and X-ray computed tomography (CT) for peri-operative margin assessment. Methodology Patients who were scheduled for the resection of a confirmed early-stage breast tumor (Tis, T1, or T2) were eligible to participate in this study. Approximately two hours before standard-of-care BCS, patients were intravenously injected with 4 MBq/kg 18F-FDG. The resected specimens were subsequently imaged by a preclinical micro-PET-CT system (β and X cube; Molecubes, Ghent, Belgium). After imaging, the resected specimens were sent to the pathology department for histopathological analysis. Margin status was post-hoc assessed by eight independent image evaluators (n=2 breast surgeons, n=2 nuclear medicine physicians, n=2 radiologists, and n=2 clinical scientists). Margins of the invasive component (cases with invasive ductal carcinoma and invasive lobular carcinoma, IDC and ILC) or the in situ component (DCIS) were considered, and a positive margin was defined as ink on tumor. The diagnostic performance was calculated based on the results of histopathological analysis as a gold standard. Results Thirty patients were included in the study: n=24 with IDC, n=4 with ILC, and n=2 with DCIS. This diagnostic technique permitted high-resolution PET-CT imaging of the resected breast tumors, enabling the identification of regions with high 18F-FDG uptake. The overall diagnostic accuracy was 89%, with the highest accuracy for IDC tumors (92%), followed by ILC (81%) and DCIS (69%). During this study, we also observed that image evaluators with more experience in interpreting specimen PET-CT images demonstrated higher diagnostic accuracy (data not shown). Moreover, the results of the breast surgeons were comparable with the results of imaging experts (radiologists and nuclear medicine physicians, data not shown). Conclusion The results of this interim analysis demonstrate that high-resolution 18F-FDG-PET-CT can facilitate peri-operative margin assessment during BCS. This technique achieves high diagnostic accuracy for IDC tumors, and may therefore reduce reoperation rates in the future. Results for ILC and DCIS are also promising. Optimization of specimen PET-CT imaging for these tumor types could further improve the diagnostic performance. Furthermore, we hypothesize that the diagnostic performance of specimen PET-CT image interpretation will further enhance by training and increasing experience of the image evaluators. Study registration This ongoing clinical study was registered in the www.clinicaltrials.gov database as NCT04343079. Table: Diagnostic performance of high-resolution 18F-FDG-PET-CT specimen imaging for margin assessment in breast malignancies.tumor typediagnostic performance (%)accuracyNPVPPVspecificitysensitivityoverall (n=30)8991899382IDC (n=24)9293939684ILC (n=4)8185908875DCIS (n=2)6980756375Abbreviations: NPV, negative predicted value; PPV, positive predicted value; IDC, invasive ductal carcinoma; ILC, invasive lobular carcinoma; DCIS, ductal carcinoma in situ. Citation Format: Menekse Göker. High-resolution 18F-FDG PET-CT for peri-operative margin assessment in early-stage breast cancer: Results of an ongoing clinical study [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-20-04.

An Investigation Using Specific Absorption Rate Analysis to Diagnose Early- stage Breast Tumor using UWB Antenna.

This work was focused on the detection of early-stage breast tumors and their location. The most frequently seen disease progression and mortality in women's lives is Breast Cancer. Because of breast cancer/tumors, the risk of mortality for women has risen exponentially. From the 2020 deadline for breast cancer, women who died from carcinoma were 123.8 cases per lac women between 2006-2010. This problem is also overcome by the early identification of the tumor using different detection procedures like X-ray mammography, computerized tomography, ultrasound imaging technique, Positron Emission Tomography (PET), Magnetic Resonance Imaging (MRI), microwave imaging. Researchers carried out multiple studies in these areas. To detect early-stage breast tumors and their location using SAR analysis. The major dielectrical difference between cancerous breast tissues and normal tissues in this technique is the microwave frequency range. The term Specific Absorption Rate (SAR) describes the amount of energy which is absorbed (W/kg) in the breast tissue. This segment illustrates the usefulness to diagnose the tumor position in the breast by means of maximum SAR value coordinates. Changes in breast and tumor size are important for the risk of diagnosis. The power absorbed in connecting with a normal breast and a tumor breast is measured and equivalent for different breast masses. The maximum SAR is also analyzed at distinct tumor locations at various frequency ranges. It is observed that max SAR coordinates are very close to the actual tumor location. So, the maximal value of SAR coordinates indicates the existence of a tumor in the breast phantom. The simulated data above strongly suggests that the Max SAR values were higher in the breast phantom with tumor as compared to the breast without tumor. With different tumor radius (3 mm and 5 mm) analyzed with different resonant frequencies like 3GHz, 4GHz and 5GHz at the actual tumor location of (0, 0, 35). Even though a model representing the real properties of breast tissue is required to assess the validation of any imaging process, so the real-time development of an equivalent breast phantom and its execution is needed.

ASPECTS EVOLUTIFS APRES TRAITEMENT CONSERVATEUR DES CANCERS DU SEIN A L’INSTITUT DU CANCER DE DAKAR. A PROPOS DE 72 CAS

Conservative treatment has become the standard treatment for early-stage breast tumors. It accounts for more than 75% of breast cancer treatments in developed countries and in our institution 6%. We studied the evolutionary aspects of breast cancer treated in a conservative modality at the Aristide Le Dantec Hospital Cancer Institute between 2000 and 2009. Objectives Describe the epidemiological profile, their evolutionary and therapeutic aspects. Methodology This work is a descriptive retrospective study of breast cancer patients selected over a 10-year period from January 2000 to December 2009 and treated conservatively at the Joliot Curie Institute in Dakar. Results We selected 72 patients out of 1100 cases treated for histologically confirmed breast cancer who had an initial conservative treatment representing 6.54%. The average age of our patients was 44.4 years. Quadrantectomy associated with axillary dissection was performed in 64 patients, or 89% of cases. Radiotherapy was performed in 53 patients, or 73.6% of cases. Twentytwo percent had locoregional recurrence (n = 16). Of the 16 patients who had recurrence, (87.5%) (n = 14) had local recurrence and 2 (12.5%) had lymph node recurrence in the internal breast chain. The average time to onset of recurrence was 17.44 months. Twenty-six percent (n =) had developed metachronous metastasis. The average time to onset of metastases was 22.63 months. The recurrences were treated by mastectomy followed by chemotherapy and radiotherapy in 8 patients, 5 patients underwent mastectomy followed by chemotherapy, 1 patient received chemotherapy alone, 1 patient received chemotherapy and palliative radiotherapy for recurrence at the the internal mammary chain and 1 patient lost to follow-up without treatment after recurrence. Metastases were treated according to different therapeutic strategies. Overall survival was 80% at 5 years and 10 years for patients with no recurrence and 37% at 5 years and 10 years for patients with recurrence. Conclusion Conservative treatment of breast cancer is possible. The results need to be improved by the generalization of mammography to discover early stages. The choice of a conservative treatment must take into account local relapse factors and diagnostic, therapeutic and follow-up means to avoid the occurrence of recurrence in order to improve patient survival.

Breast Tumor Malignancy Classification using Smartphone Compression-induced Sensing System and Deformation Index Ratio.

Smartphone-based compression-induced sensing system uses the light diffusion pattern to characterize the early-stage breast tumor noninvasively. The system is built on a smartphone and cloud platform to capture, transfer, and interface with the user. The compressed tissue's deforming pattern creates distinctive tactile images due to the size and hardness of the tumor. From the compression-induced images, we estimate the size of the tumor using projection analysis and the tumor's malignancy using the tissue deformation index ratio. Deformation index ratio is based on the changes of a healthy region over the tumorous region. By using the projection analysis, the human patient tumor size estimation resulted in 52.3% of the average error. For a small number (seven) of the feasibility test, the tumor's malignancy was classified based on the deformation index ratio with 67.0% of sensitivity and 100% specificity.

Changes in black-white disparity in receipt of guideline concordant treatment for women with early-stage breast tumors.

e19089 Background: Racial disparities are well documented in the treatment of women with breast cancer. Whereas clinical treatment guidelines are established for early stage breast tumors, disparities in guideline-concordant (GC) treatment may influence variation in breast cancer outcomes. As such, we investigated differences in the receipt of GC treatment for black and white women with early-stage breast cancer between 2008 and 2016. Methods: We evaluated the Surveillance, Epidemiology, and End Results Registry (SEER) Incidence database for black and white women aged 20-64 years with stage I / stage II breast tumors. Primary analyses investigated associations between race and receipt of GC treatment in three chronological periods. Potential driving factors for trends in receipt of guideline-concordant treatment were also assessed. Results: Among 145,561 women diagnosed with early stage breast tumors, overall receipt of GC care decreased from 84.58% in 2008 to 83.89% in 2016. In period 1 (2008-2010), there was a prominent disparity between black and white women (81.89% black vs 86.24% white; p &lt; 0.001). By period 3 (2014-2016), GC care increased for black women (84.29%) but decreased for white women (84.82%), eliminating the disparity (p = 0.276). Multivariate logistic regression on changes in the black-white disparity across the study period showed increased receipt of GC treatment for black women relative to white women in period 2 (2011-2013, odds ratio [OR], 1.202; 95% confidence interval [CI], 1.075 – 1.344) and period 3 (OR, 1.376; 95% CI, 1.212 – 1.505). For black women undergoing breast conserving surgery, administration of radiation therapy increased from 44.01% in period 1 to 50.64% in period 3 (p &lt; 0.001) and was a prominent driver for increased GC care. Conclusions: Overall receipt of guideline-concordant treatment decreased from 2008-2016. Black women experienced a substantial increase in receipt of guideline-concordant care in period 2 (2011-2013) and period 3 (2014-2016) relative to white women. Delivery and completion of radiation therapy with breast conserving surgery increased the likelihood of receipt of guideline-concordant treatment.

Distinct genomic profiles of 589 Chinese early-stage breast cancer.

552 Background: Extensive efforts by The Cancer Genome Atlas (TCGA) Network had provided much of our current understanding of the molecular profile of various solid tumors including breast tumors; however, Asian patients were underrepresented in this cohort. In this study, we aimed to elucidate the comprehensive genetic alteration profile of early-stage breast tumors among Chinese patients. Methods: Surgical tissue samples from 589 Chinese women with stage I-III breast cancer with various histology and molecular subtype consecutively diagnosed at Guangdong Provincial People’s Hospital were sequenced using a panel targeting 520 cancer-related genes spanning 1.64Mb of the human genome. Clinical and genomic data from our cohort was compared with publicly-available data from 1,046 stage I-III breast cancer patients from the TCGA dataset. Results: Based on the analysis of the genetic alteration profile from our cohort, at least one genetic alteration was observed from 98% of the tumor samples, with TP53 (47%), PIK3CA (45%), ERBB2 (30%), and CDK12 (18%) as the most commonly altered genes. The most common genetic alteration types were copy number amplification (43.6%) and missense mutations (36.8%). As compared with the TCGA dataset, our cohort is mostly composed of women 50 years and younger (59.1% vs. 30.4%, P&lt; 0.001), with significantly fewer patients with lobular carcinoma histology (2.4% vs. 19.0%, P&lt; 0.001), and significantly more patients with pathologic stage I tumors (23.3% vs. 17.3%, P= 0.012). Consistently, genetic alterations detected from our cohort affected genes involved in PI3K (63% vs. 56%, P= 0.009) and cell cycle (23% vs. 35%, P&lt; 0.001) pathways, with statistically different genetic alteration rates as compared with the TCGA dataset. Comparison of genetic alteration profile between the two cohorts revealed that our cohort had more frequent genetic alterations in genes including PIK3CA ( P&lt; 0.001), TP53, particularly in hotspot mutations Q192* ( P&lt; 0.001) and A307V/del ( P= 0.02), and ERBB2 amplification ( P&lt; 0.001). Conclusions: Our study contributes to the understanding of the key pathways and specific genetic alterations harbored by Chinese patients with early-stage breast cancer that could potentially be developed as markers of treatment response to targeted therapeutics.

Targeting Pro-Tumoral Macrophages in Early Primary and Metastatic Breast Tumors with the CD206-Binding mUNO Peptide.

M2-like tumor-associated macrophages (M2 TAMs) play important roles in the resistance of tumors to immunotherapies. Selective depletion or reprogramming of M2 TAMs may sensitize the nonresponsive tumors for immune-mediated eradication. However, precision delivery of payloads to M2 TAMs, while sparing healthy tissues, has remained an unresolved challenge. Here, we studied the application of a short linear peptide (CSPGAK, "mUNO") for the delivery of molecular and nanoscale cargoes in M2 TAMs in vitro and the relevance of the peptide for in vivo targeting of early-stage primary breast tumors and metastatic lung foci. First, we performed in silico modeling and found that mUNO interacts with mouse CD206 via a binding site between lectin domains CTLD1 and CTLD2, the same site previously demonstrated to be involved in mUNO binding to human CD206. Second, we showed that cultured M2 macrophages take up fluorescein-labeled (FAM) polymersomes conjugated with mUNO using the sulfhydryl group of its N-terminal cysteine. Pulse/chase studies of FAM-mUNO in M2 macrophages suggested that the peptide avoided lysosomal entrapment and escaped from early endosomes. Third, our in vivo studies with FAM-mUNO demonstrated that intraperitoneal administration results in better pharmacokinetics and higher blood bioavailability than can be achieved with intravenous administration. Intraperitoneal FAM-mUNO, but not FAM-control, showed a robust accumulation in M2-skewed macrophages in mouse models of early primary breast tumor and lung metastasis. This targeting was specific, as no uptake was observed in nonmalignant control organs, including the liver, or other cell types in the tumor, including M1 macrophages. Collectively, our studies support the application of the CD206-binding mUNO peptide for delivery of molecular and nanoscale cargoes to M2 macrophages and manifest the relevance of this mode of targeting primary and metastatic breast tumors.

Abstract P4-04-04: Loss of epigenetic lineage fidelity with age primes breast epithelia for malignant transformation

Abstract Background: Aging is the largest risk factor for breast cancer, with 80% of new cases diagnosed in women over 50y. However, the molecular mechanism underlying age-associated cancer susceptibility is still not well understood. While aging-associated phenotypes in a number of tissues have been correlated with transcriptional and epigenetic changes, few studies have looked at epigenetic regulation at the resolution of individual cell populations. The bilayered epithelia of the mammary gland is composed of an apical layer of secretory luminal epithelial cells (LEP) surrounded by a basal layer of contractile and tumor suppressive myoepithelial cells (MEP). Breast cancer cells of origin are thought to reside mainly in the luminal or supra-basal regions directly adjacent to the apical surfaces of the MEP. Thus, an understanding of how LEP and MEP interact and maintain lineage-specific expression, and how this is dysregulated with age may elucidate a potential mechanism regulating increased breast cancer susceptibility with age. Previously, we showed that loss of lineage fidelity of LEP and MEP is a key feature of aging. Moreover, this loss of lineage fidelity can be imposed by old MEP onto young LEP in a cell non-autonomous manner (Miyano &amp; Sayaman, et al, Aging 2017). Methods: We present a detailed analysis of genome-wide DNA methylation (Infinium Human Methylation 450K) changes at lineage-specific resolution from FACS-sorted MEP and LEP populations isolated from finite lifespan human mammary epithelial cells (HMEC) derived from reduction mammoplasties of younger (&amp;lt;30y) and older women (&amp;gt;55y). Results: In younger epithelia, maintenance of lineage fidelity is associated with lineage-specific differential methylation (DM) of CpG sites at key regulatory regions. We find that loss of lineage fidelity is recapitulated in genome-wide DNA methylation, with loss of DM at canonical proximal regions associated with transcriptional regulation, specifically annotated CpG island groups, enhancer regions, and CTCF binding sites thought to be involved in chromatin remodeling. Strikingly, DM changes with age occur almost exclusively in LEP population, with older LEP acquiring more MEP-like DNA methylation patterns at the dysregulated sites. Aged LEP and MEP also exhibit significant variability in DNA methylation at both hypo- and hypermethylated regions that show little variance in young samples. Furthermore, predictions of physiologic age using Horvath’s clock show MEP to have older physiologic ages than their isogenic LEP counterparts. Titus et al, found that DM regions (DMRs) between early stage breast tumors and normal-adjacent breast samples in The Cancer Genome Atlas (TCGA) are significantly more enriched in the LUMA, LUMB and HER2 than the Basal molecular subtypes (Sci Reports, 2017). We show that CpG sites associated with the luminal-subtype DMRs are lineage specific and that a significant fraction of the epigenetic changes that occur in these early stage breast cancers are already present during aging. Conclusions: The lineage bias of age-specific of DNA methylation changes suggests that epithelial lineages age via different mechanisms. That LEP are significantly more differentially regulated via DNA methylation with age than MEP raises possibilities that DNA methylation affects aging-associated breast cancer risk in a lineage-specific manner. This lineage bias, along with age-specific changes in breast composition, may underlie the differences in incidence rates of breast cancer subtypes with age and the prevalence of ER+ luminal-subtype breast cancers in women post-menopause. We suggest that these epigenetic changes in the luminal lineage may be priming events that make breast cells more vulnerable to further dysregulation that leads to cancer. Citation Format: Rosalyn W. Sayaman, Masaru Miyano, Mark A. LaBarge. Loss of epigenetic lineage fidelity with age primes breast epithelia for malignant transformation [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 P4-04-04.

Abstract P6-04-17: Role of HOXB13 in modulating estrogen signaling in breast cancer cells

Abstract Background: The ratio of HOXB13 (H) and IL17BR (I) gene expression levels in hormone receptor-positive (HR+) primary early stage breast tumors define a predictive index (H/I) that, combined with the Molecular Grade Index (MGI), constitute the Breast Cancer Index (BCI). The BCI score (MGI+H/I prognostic model) provides individualized risk of overall (0-10 years) and specifically late (5-10 years) distant recurrence, and BCI (H/I) predicts the benefit of extended endocrine therapy (EET). Hormone receptor responses are dependent on genome-wide hormone receptor binding patterns. In hormone-dependent cancers, nuclear receptor binding patterns are frequently reprogrammed by other transcription factors. For example, in breast cancer cells, GATA3 has been shown to reprogram estrogen receptor (ER) binding and to alter gene expression. Similarly, HOXB13 was previously shown to reprogram genome-wide binding of the androgen receptor (AR) during prostate cancer tumorigenesis. To assess whether HOXB13 expression is associated with regulation of ER binding, global ER binding data in highly HOXB13-expressing, tamoxifen-resistant breast cancer cells to the ER binding pattern in HOXB13-low, tamoxifen-sensitive breast cancer cells were compared. The study objective was to understand whether HOXB13 contributes to ER reprogramming in early-stage breast cancer. Methods: Global ER, HOXB13 and FOXA1 DNA binding patterns and the enhancer activation mark histone H3 lysine 27 acetylation (H3K27ac) were characterized in BT474 cells by chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-Seq). To compare ER binding in highly HOXB13-expressing, tamoxifen-resistant BT474 cells to published ER binding data in HOXB13-low, tamoxifen-sensitive MCF-7 breast cancer cells, we downloaded published ER, FOXA1, GATA3 and H3K27ac ChIP-seq data from MCF-7 cells. After aligning reads with Bowtie2, peak calling and data integration was performed using HOMERv4.10. Results: Analysis of highly HOXB13-expressing BT474 cells identified a large number of HOXB13 binding sites, which overlapped with approximately half of the ER binding sites in these cells. Many of these binding sites were co-bound by FoxA1, a general pioneer factor that has been shown to also colocalize with HoxB13 at reprogrammed AR binding sites in human prostate tumors. Interestingly, most of these locations are distinct from the genomic binding sites of ER in HOXB13-low MCF-7 cells, where ER frequently binds to MCF-7-specific sites in conjunction with GATA3. Conclusion: Our preliminary evidence suggests that HOXB13 influences the estrogen receptor binding pattern in human breast cancer cells and that ER binding sites differ between HOXB13 high- and low-expressing cell lines, which may underlie their differential responses to endocrine therapy. These results will be compared to ongoing experiments assessing the effects of conditional expression or loss of HOXB13 on ER binding and function in HOXB13-negative and HOXB13-positive cells, respectively. Citation Format: Kai Treuner, Michael Hayes, Christopher Benner, Catherine Schnabel, Sven Heinz. Role of HOXB13 in modulating estrogen signaling in breast cancer cells [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 P6-04-17.

A thermoacoustic imaging system with variable curvature and multi-dimensional detection adapted to breast tumor screening

Microwave-induced thermoacoustic (TA) imaging is efficient in detecting anomalous absorption areas, especially breast tumors. The operation of previous imaging systems has generally suffered from complexity, and early-stage tumors could be missed because of signal loss. In this paper, we report a variable-curvature and multi-dimensional screening method realized by combining a redesigned adaptive algorithm and an integrated TA breast screening system (ITBCS) comprising a flexible detector and a traction device designed for multi-axis rotation. In contrast to conventional TA imaging systems, the proposed system improves the ability to identify early-stage breast tumors, with a simplified system and operational process. Variable curvature is achieved by the flexible characteristics of the detector, which achieves good shape adaptation between the tissue and the detector. Moreover, multi-dimensional scanning is achieved by the multi-axis traction device and meets the requirements for angle transformation and dynamic imaging. The experimental results indicate that ITBCS exhibits a good performance in detecting breast tumors. Faster screening makes inspection more efficient, thereby improving clinical applicability and providing a convenient facility for large-scale physical examination.