Purpose
This study aimed to elucidate the clinicopathological characteristics of breast tumors with homologous recombination deficiency (HRD) and the sensitivity to neoadjuvant paclitaxel followed by ...fluorouracil, epirubicin, and cyclophosphamide (P-FEC).
Methods
Tumor biopsy samples obtained before P-FEC from 141 patients with stages II–III breast cancer including the luminal (
n
= 76), luminal-HER2 (
n
= 13), HER2 (
n
= 17), and triple-negative (TNBC,
n
= 35) subtypes were subjected to assay for HRD score using the OncoScan CNV FFPE Assay Kit. HRD score was a simple sum of NtAI, LOH, and LST (cutoff, 42). TNBCs were also subjected to the gene expression assay using the Affymetrix microarray (U133 plus 2.0) and to the
BRCA1
promoter methylation assay using the methylation-specific real-time PCR.
Results
Of the 141 breast tumors, 45 samples (32%) had high HRD scores and were associated with high histological grade (
P
= 0.001), negative progesterone receptor (
P
= 0.018), high Ki67 index (
P
= 0.032), and
BRCA1
promoter methylation (
P
= 3.6e−07). The proportion of tumors with high HRD scores was significantly higher in the TNBC subtype than the others (
P
= 0.006). In the TNBC subtype, but not the others, high HRD scores were significantly (
P
= 0.001) associated with a low pathological complete response rate to P-FEC. Among the molecular TNBC subtypes, a majority of tumors belonging to the basal-like 1, immunomodulatory, mesenchymal, mesenchymal stem-like, but not luminal androgen receptor (LAR), subtypes had high HRD scores.
Conclusions
Approximately one-third of sporadic breast tumors show a high HRD score, indicating the presence of homologous recombination dysfunction, and they are characterized by biologically aggressive phenotypes, most commonly in the TNBC subtype, and less sensitive to P-FEC.
Abstract
AIM: We have previously reported that the serum concentration of anti-HER2 autoantibodies (HER2-AAb) is associated with favorable outcomes in patients with invasive breast cancer (Tabuchi et ...al., Breast Cancer Res Treat 157:55-63, 2016). This study aimed to investigate the prognostic impact of HER2-AAb by examining the tumor microenvironments for humoral immunity.
PATIENTS AND METHODS: From 500 consecutive patients with invasive breast cancer (Ibid.), we selected those whose log-transformed HER2-AAb values were above mean + 2SD (high HER2-AAb group, N = 33) or below mean − 2SD (low HER2-AAb group, N = 20). Tumor formalin-fixed paraffin-embedded (FFPE) samples and regional lymph node FFPE samples prepared from the patients’ surgical specimens were subjected to immunohistochemistry. Tumor-infiltrating immune cells (ICs) localized inside the tumor, in the adjacent stroma, and in the tumor stroma were separately counted. Expression of some proteins in tumor cells (TCs) was evaluated by the percentages of tumor cells showing presence of those proteins.
RESULTS: We confirmed in the selected patients that the recurrence-free interval of the high HER2-AAb group was significantly longer than that of the low HER2-AAb group (log-rank P = 0.017; hazard ratio = 0.12). Tumor-infiltrating CD20-positive ICs (intratumoral + adjacent stromal ICs, P < 0.001), IGKC-positive ICs (intratumoral + adjacent stromal ICs, P = 0.023), and CXCL13-positive ICs (total ICs, P = 0.044) were significantly greater in the high HER2-AAb group than in the low HER2-AAb group. CD4-positive ICs in B-cell follicles of the regional lymph nodes were also significantly greater in the high HER2-AAb group than in the low HER2-AAb group (P = 0.026). Tumor-infiltrating PD-L1-positive ICs, CD8-positive ICs, FOXP3-positive ICs, as well as CD8/FOXP3 ratios, were not significantly different between the two groups. The expression of CXCL13, HLA-A/B/C, and PD-L1 in TCs was not significantly different between the two groups. The repertoire of B-cell receptors in ICs was not significantly different between the two groups but was considered to be skewed. HER2 expression and gene amplification and the existence of HER2 missense mutations were similar between the two groups.
CONCLUSIONS: Increased HER2-AAb concentration was associated with enhanced humoral immunity in tumor microenvironments. Our findings indicate that a patient subpopulation with enhanced humoral immunity protecting against breast cancer recurrence exists. The recognition site of HER2-AAb may not be located in the HER2 polypeptide, but may be located in modifying factors, such as sugar chains.
Citation Format: Masafumi Shimoda, Yasufumi Sato, Yoshiaki Sota, Tomohiro Miyake, Tomonori Tanei, Naofumi Kagara, Yasuto Naoi, Kenzo Shimazu, Seung Jin Kim, Shinzaburo Noguchi. Association of autoantibodies to HER2 with tumor microenvironments for humoral immunity and prognosis in patients with breast cancer abstract. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1604.
The pathologic feature of intraductal papillomas is defined as a papillary structure composed of a fibrovascular stromal core lined by luminal epithelial cells and myoepithelial cells. We used ...droplet digital PCR for the mutational analysis of AKT1 (E17K) and PIK3CA (H1047R, E542K, and E545K) in 60 papillomas. AKT1 and PIK3CA mutations were detected in 12 (20%) and 17 (28%) of the papillomas, respectively. In five tumors harboring mutations, mutational analysis of AKT1 or PIK3CA was performed separately using luminal epithelial cells and myoepithelial cells sorted using anti–cytokeratin 19 antibody and anti–α smooth muscle actin antibody. The two types of cells from a given papilloma had the identical mutation. Three patients with the PIK3CA mutation–positive papilloma developed breast cancers at the resection site of the papilloma, but none of these subsequent breast cancers had the PIK3CA mutation. These results indicate that a papilloma stems from a bipotent progenitor cell that contains the AKT1 or PIK3CA mutation and proliferates and differentiates to form the papilloma. Papilloma can be a risk factor for developing breast cancer but is unlikely to be its obligate precursor.
Developing novel strategies against treatment-resistant triple negative breast cancer (TNBC) cells remains a significant challenge. The ErbB family, including epidermal growth factor receptor (EGFR), ...plays key roles in metastasis, tumorigenesis, cell proliferation, and drug resistance. Recently, these characteristics have been linked to a small subpopulation of cells classified as cancer stem cells (CSC) which are believed to be responsible for tumor initiation and maintenance. Ixabepilone is a new generation microtubule-stabilizing agent, which has been expected to be more efficacious than conventional taxanes. Here we aim to investigate whether the EGFR monoclonal antibody Cetuximab, in combination with Ixabepilone, is more effective in eliminating CSC populations compared to chemotherapy alone in TNBC.
Representative TNBC cell lines (MDA-MB-231 and SUM159) were used to evaluate breast CSC populations. We used fluorescence-activated cell sorter analysis (CD44(+) and CD24(-/low), or Aldefluor(+)) and a self-renewal assay called mammosphere formation efficiency (MSFE) to measure CSC population size after treatment with Cetuximab, or Cetuximab plus Ixabepilone in vitro.
Although there was no significant decrease in cell viability, Cetuximab reduced MSFE and the CSC population in breast cancer cells in vitro and in vivo through inhibition of autophagy. Also, SUM159 and MDA-MB-231 orthotopic tumors demonstrated partial response to Centuximab or Ixabepilone monotherapy; however, the effect of the combination treatment was significant only in SUM159 tumors (p <0.0001), when compared to Ixabepilone alone.
Overall, our findings demonstrate that EGFR-targeted therapy by Cetuximab effectively reduces the CSC population in TNBC tumors. However, combination therapy with Ixabepilone may be effective only in a small subset of TNBCs, warranting further investigation of alternative approaches to target multiple pathways for TNBC treatment.
10
Background: The IRSN-23 model uses DNA microarray analysis of tumor tissue to stratify patients into highly sensitive (Gp-R) or less sensitive (Gp-NR) chemotherapy groups based on immune-related ...gene expression. In a previous report (n=901), Gp-R showed a 39% pCR rate, while Gp-NR had 11% (P=4.98E-23; Sota, Y., et al., Annals of Oncology 25.1 (2014): 100-106). This study aimed to evaluate the reproducibility of the IRSN-23 model in prospective independent validation cohorts and investigate its clinical significance and impact on breast cancer subtype classification. Methods: Tumor tissues were obtained from 143 breast cancer patients undergoing preoperative chemotherapy at Osaka University Hospital (OUH). Patients were classified into Gp-R or Gp-NR using the IRSN-23 model, and its ability to predict pathological complete response (pCR) was assessed. Results were validated with independent public datasets (n=965; breast cancer (n=919) and rectal cancer (n=46)). Results: In the OUH prospective cohort, the pCR rate was significantly higher in the Gp-R group (29.3% (11/41)) than in the Gp-NR group (1.4% (1/71)) without trastuzumab (P=1.70E-5). In all prospective validation cohorts without anti-HER2 therapy, the pCR rate in the Gp-R group was significantly higher than in the Gp-NR group. Pooled analysis of the prospective validation set revealed higher pCR rates in the Gp-R group than in the Gp-NR group, both without (Gp-R 41.9% (104/248) vs. Gp-NR 13.6% (50/367), P=4.0E-15) and with (Gp-R 48.3% (57/118) vs. Gp-NR 35.5% (66/186), P=0.03) anti-HER2 therapy (Table). Collaborative analyses of IRSN-23 Immune Score (IS) and OncotypeDx Recurrence Score (RS) suggested the identification of a highly sensitive chemotherapy group (pCR%; IS>-25+RS>25: 21.6% vs. IS ≤ 25 + RS > 25: 2.9%, P=1.13E-04). PAM50 as an attack factor and IRSN-23 as a defense factor capture the tumor microenvironment, suggesting a new subtype classification of breast cancer. Conclusions: This study offers prospective validation of the IRSN-23 model in predicting chemotherapy efficacy, exhibiting high reproducibility. The findings indicate the IRSN-23 model is clinically valuable and propose new breast cancer subtype classification. These results hold significant implications for personalized treatment strategies.Table: see text
We have shown that “Click-to-sense” (CTS) assay based on the visualization of cancer cells by fluorescence probe targeted for acrolein is useful for differentiating between the malignant and benign ...lesions of the breast. In the present study, we aimed to apply CTS assay to the examination of the simulated surgical margins, being compared with frozen section (FS) analysis.
The simulated surgical margin samples (n = 300) were obtained from 1 to 2 cm distant sites from the tumor margin in the mastectomy specimens of breast cancer patients, and divided into the training (n = 150) and validation (n = 150) set. The samples were subjected to CTS assay, subsequently to FS analysis and finally to permanent section (PS) analysis.
Diagnostic accuracy of the CTS assay and FS analysis was evaluated in the examination of the simulated surgical margin status finally determined by the PS analysis. In the training set, sensitivity, specificity, and accuracy was 89.3%, 98.4%, and 96.7% for the CTS assay and 89.3%, 98.4%, and 96.7% for the FS analysis. In the validation set, sensitivity, specificity, and accuracy was 93.3%, 98.3%, and 97.3% for the CTS assay, and 93.3%, 99.2%, and 98.0% for the FS analysis.
The CTS assay is as accurate as the FS analysis in the examination of the simulated surgical margins in breast cancer patients, and it seems to have a potential to replace the FS analysis for the intra-operative examination of surgical margins in breast-conserving surgery since it is less labor-intensive and more time-saving than the FS analysis.
•Click-to-sense (CTS) assay can visualize cancer cells in live tissue in short time.•It is important to examine surgical margin in breast conserving surgery.•CTS assay was applied to examination of simulated surgical margin samples.•CTS assay was as accurate as frozen section (FS) analysis.
Background/Aim: Magnetic resonance imaging (MRI)-guided breast biopsy is a complex and time-consuming procedure. This study aimed to clarify the factors that affect the duration of the procedure. ...Patients and Methods: Twenty-eight examinations performed at our institute for 27 lesions detected solely on MRI were analyzed. The correlations between the clinicopathological factors and duration of the procedure were estimated. Results: The needle guidance method was the only factor that significantly affected the duration of the MRI-guided vacuum-assisted breast biopsy (VAB) (p=0.012). The use of a computer-aided detection (CAD) system with grid breast compression plates had significantly shorter durations (62±12 min) than the manual calculation of coordinates with pillar-type compression plates (76±13 min). Conclusion: This preliminary study showed that the use of a CAD system might shorten the duration of MRI-guided VAB.
The restriction enzyme-based digital methylation-specific polymerase chain reaction (RE-dMSP) assay is useful for diagnosing sentinel lymph node (SN) metastasis in patients with breast cancer, by ...detecting tumor-derived methylated Ras association domain-containing protein 1 (RASSF1A). In addition, this assay has high concordance (95.0%) with one-step nucleic acid amplification (OSNA). The present study aimed to perform RE-dMSP using OSNA lysate from more patients and to re-evaluate its clinical usage. Overall, 418 SNs from 347 patients were evaluated using both OSNA and RE-dMSP. The concordance rate was 83.3% (348/418). RASSF1A methylation of the primary tumors was negative in 36 patients. When these patients were excluded, the concordance rate improved to 88.2% (330/374). Of the 79 OSNA-negative cases, 19 were RE-dMSP-positive, although all were positive for cytokeratin 19 expression in the primary tumor, suggesting that RE-dMSP can detect tumor-derived DNA with a higher sensitivity. The percent of methylated reference of the breast tumors showed a wide variety in the 16 OSNA-positive/RE-dMSP-negative cases, and such variability of methylation could have affected the results in these patients. In conclusion, although RE-dMSP can diagnose SN metastasis with high sensitivity and accuracy, and can be a supplementary tool to OSNA in breast cancer, RE-dMSP showed certain discordance with OSNA and critically depended on the absence or heterogeneity of DNA methylation in breast tumors. Further research is expected to develop an assay targeting other DNA alterations, such as mutations. Key words: SN, molecular diagnosis, RASSF1A promoter methylation, OSNA, breast cancer, dPCR
We aimed to validate the cosmetic utility of addition of nipple-areola recentralization (NAR) to rotation flap according to nipple tumor distance (NTD) as a volume displacement technique after breast ...conserving surgery (BCS) for lower-outer and upper-inner breast cancers. Twenty breast cancer patients who had been treated with rotation flap with (Group 1; n = 6) or without (Group 2; n = 14) NAR after BCS for lower-outer or upper-inner located tumors, and those who had undergone BCS without oncoplastic surgical technique for tumors in the same area (Control group; n = 43), were retrospectively investigated. Cosmetic outcome was evaluated using Harvard scale and/or BCCT.core. As a result, the ratio of patients categorized as excellent/good was 83% in Group 1 and 93% in Group 2, respectively, and there was no significant difference between them (P = 0.521). In addition, Group 1 + 2 showed a significantly higher ratio of patients classified as excellent/good than the control group (90% vs. 56%; P = 0.009). After adjustment of clinical background parameters using propensity score matching analysis between Group 1 + 2 and the control group, 12 pairs with similar background factors were matched. Among them, Group 1 + 2 showed a higher ratio of patients categorized as excellent/good than the control group (92% vs. 42%; P = 0.034). In conclusion, addition of NAR to rotation technique according to NTD may enable us to perform a volume displacement after BCS for lower-outer or upper-inner located tumors irrespective of NTD without sacrificing postoperative breast appearance.
Rapidly growing cancer cells have increased levels of intracellular polyamines compared to normal, healthy tissues. Based on the selective reactivity of glycine propargyl esters, probes were ...synthesized that show evidence for selective polyamine reactivity, which was then applied for selective cancer cell imaging studies.
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