Low Hormone Receptor Expression Research Articles

Clinical significance of circulating tumor DNA in neoadjuvant-treated HER2-negative luminal B breast cancer.

e12616 Background: Circulating tumor DNA (ctDNA) holds significant value in the comprehensive management of breast cancer (BC) patients, whereas long-standing questions have been raised concerning the practical utility of ctDNA in HR+/HER2- subtype. Herein, we evaluated the clinical activity of ctDNA in luminal B (HR+/HER2-, with either Ki67≥20% and/or PR<20%) BC patients receiving neoadjuvant therapy. Methods: Patients received neoadjuvant chemotherapy (NCT) or neoadjuvant endocrine therapy combined with CDK4/6 inhibitors (NET) were enrolled. Tumor tissue and plasma samples were collected and subjected to sequencing. Blood was collected for ctDNA analysis before treatment (T0), 2-3 weeks after treatment (T1), at the midpoint of neoadjuvant treatment (T2), prior to surgery (S), and postoperatively (PO). The tumor tissues were biopsied or surgically excised at T0, T1, and S to assess somatic mutation profiling and pathological markers. NGS assays were performed with a custom-designed panel that covered 769 cancer-related genes. Results: As of Jan 7, 2024, 448 tumor tissue and plasma samples from 80 BC patients undergoing NCT (n = 57) or NET (n = 23) were sequenced and analyzed. At T0, positive ctDNA was detected in 62.8% (49/78, another two failed quality control) of the patients. Furthermore, ctDNA detection and the higher concentration had a significant correlation with clinicopathologic characteristics that confer aggressiveness, including larger tumor size, more lymph node metastases, and lower hormone receptor expression. There was also a certain relativity between ctDNA positivity and tumor grading as well as MammaPrint index ( P < 0.1). With the progression of neoadjuvant therapy, the ctDNA positive rate decreased over time (T1: 32/80, 40.0%; T2: 11/61, 18.0%; S: 7/65, 10.8%; and PO: 6/64, 9.4%). Although no correlation was found between pretherapeutic Ki67 expression and T0 ctDNA status ( P = 0.871), the Ki67 expression in early cycles of neoadjuvant therapy (T1) was significantly higher in patients with inter-therapeutic detectable ctDNA. Moreover, the expression and decline of Ki67 after neoadjuvant therapy were significantly correlated with ctDNA positivity at T2, S, and PO. Among the patients who had undergone surgery, 8 patients were classified as RCB 0/1 and the rest 57 as RCB 2/3. All patients who exhibited RCB 0/1 were ctDNA negative at S and PO. Compared to RCB 2/3, patients with RCB 0/1 tumors showed a significant increase in ctDNA concentration at T0 ( P = 0.027). Subgroup analysis revealed similar trends in ctDNA status and concentration changes in the NCT group and NET group. Conclusions: We reported an unprecedented baseline ctDNA positivity in luminal B BC, and the ctDNA levels were closely associated with the clinicopathologic characteristics of patients. As the research progresses, more predictive and prognostic information is expected to be discovered. Clinical trial information: NCT05649475 .

Clinical, pathological, and comprehensive molecular analysis of the uterine clear cell carcinoma: a retrospective national study from TMRG and GINECO network

BackgroundUterine clear cell carcinomas (CCC) represent less than 5% of uterine cancers. Their biological characteristics and clinical management remain uncertain. A multicenter study to explore both clinical and molecular features of these rare tumors was conducted.MethodsThis multicenter retrospective national study was performed within the French TMRG (Rare Gynecologic Malignant Tumors) network. Clinical data and, when available, FFPE blocks were collected. Clinical features, treatments, and outcome (progression-free survival (PFS) and overall survival (OS)) were analyzed and correlated to the protein (tissue micro-array), RNA (Nanostring nCounter® technology), and DNA (array-Comparative Genomic hybridization and target-next generation sequencing) levels using the tumor samples available.ResultsSixty-eight patients with uterine CCC were enrolled, 61 from endometrial localization and 5 with cervix localization. Median age at diagnosis was 68.9 years old (range 19–89.7). Most tumors were diagnosed at an early stage (78% FIGO stage I–II). Hysterectomy (performed in 90%) and lymph node dissection (80%) were the most frequent surgical treatment. More than 70% of patients received external beam radiotherapy and 57% received brachytherapy. Nearly half (46%) of the patients received chemotherapy. After a median follow-up of 24.7 months, median PFS was 64.8 months (95 CI [5.3–124.4]) and median OS was 79.7 (IC95 [31.0–128.4]). Low hormone receptor expression (13% estrogen-receptor positive), frequent PI3K pathway alterations (58% PTEN loss, 50% PIK3CA mutations), and P53 abnormalities (41%) were observed. Mismatch repair deficiency was identified in 20%. P16 expression was associated with shorter PFS (HR = 5.88, 95 CI [1.56–25], p = 0.009). Transcriptomic analyzes revealed a specific transcriptomic profile notably with a high expression of immune response-associated genes in uterine CCC displaying a very good overall prognosis.ConclusionsUterine CCC reported to be potentially MSI high, hormone receptors negative, and sometimes TP53 mutated. However, some patients with immune response-associated features and better prognosis may be candidate to treatment de-escalation and immunotherapy.

Abstract P4-02-17: Impact of low hormone receptor expression on neoadjuvant chemotherapy response and patterns of care in early-stage HER2-negative breast cancer: a US National Cancer Database analysis

Abstract Background: Hormone receptor (HR) low (1-10%) HER2-negative breast cancer (BC) is emerging as a distinct subtype with similarities in clinical outcomes to triple-negative BC. However, there is a lack of consensus on treatment recommendations for chemo-immunotherapy and endocrine therapy in this subset. Here, we present results from a US National Cancer Database (NCDB) analyses of patients with HER2-negative BC evaluating response to neoadjuvant chemotherapy (NAC) and patterns of care by HR expression. Methods: Patients with stage I-III HER2-negative BC diagnosed in 2018 were identified in NCDB, a nationwide oncology outcomes database in the US. Quantitative HR expression was unavailable prior to 2018. Data were categorized into four groups by estrogen receptor (ER) and progesterone receptor (PR) expression: ER< 1% & PR< 1% (HR-Neg), ER 1-10% and/or PR 1-10% (HR-Low), ER >11-30% and/or PR>11-30% (HR-Int), ER> 30% and/or PR > 30% (HR-High). Those with undocumented HR status (3%) or without curative intent surgery (5%) were excluded. The primary outcome was pathologic complete response (pCR) by HR expression in those undergoing neoadjuvant chemotherapy. Key secondary objectives included assessment of clinicopathologic characteristics and practice patterns. The categorical variables were compared between the four groups using a Chi-square test. Age was compared using a Kruskal-Wallis test. Results: Out of 104,205 incident cases, 2541 (2.4%) were HR-Low and 1241 (1.2%) were HR-Int. Significant differences were found between HR groups with higher grade, clinical stage, and Ki-67 in HR-Low vs. HR-Int or HR-High groups (Table 1). Patients with HR-Low and HR-Int BC were more likely to receive chemotherapy than HR-High (74%, 70% vs. 20%; p < 0.001) and the use of adjuvant endocrine therapy correlated with HR expression. Only half of patients in the HR-Low group received any endocrine therapy compared to higher rates in the HR-Int and HR-High groups (52% vs. 74%, 92%; p< 0.001). pCR rates in those receiving neoadjuvant chemotherapy were significantly different by HR status, with higher pCR rates in HR-Low vs. HR-High groups (p< 0.001) (Table 2). NAC utilization significantly differed between groups. A higher proportion of patients with HR-Low BC received NAC than other HR-positive groups (p < 0.001). Additionally, there was an increased use of NAC in patients with HR-Low BC treated at academic vs. community cancer centers (p< 0.001). Conclusions: This is one of the largest real-world analyses comparing key differences in biology and practice patterns of HR-Low, HER2-negative BC. Consistent with prior studies, we report HR-Low BC to be a rare and distinct subtype with higher pCR rates compared to HR-High BC. Practice patterns show wide variability in utilization of neoadjuvant chemotherapy and endocrine therapy for these patients. Future studies should address this disparity and enhance representation of patients with HR-Low BC in clinical trials to improve long-term outcomes. Table 1: Patient demographics Table 2: Neoadjuvant chemotherapy response amongst differing HR expression levels Citation Format: Dionisia Quiroga, Michael Grimm, Julie Stephens, Kai Johnson, Nicole Williams, Preeti K. Sudheendra, Mathew A. Cherian, Daniel Stover, Ashley C. Pariser, Margaret Gatti-Mays, Robert Wesolowski, Jose G. Bazan, Sasha Beyer, Ko Un Park, Bridget Oppong, Bhuvaneswari Ramaswamy, Julia White, Sachin R. Jhawar, Sagar Sardesai. Impact of low hormone receptor expression on neoadjuvant chemotherapy response and patterns of care in early-stage HER2-negative breast cancer: a US National Cancer Database analysis [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P4-02-17.

Impact of Detection Mode in a Large Cohort of Women Taking Part in a Breast Screening Program.

The aim of this study was to evaluate the existing survival rate and clinical-pathological differences among patients with breast cancer detected by mammographic screening. This multicenter cohort study examined 1,248 patients who took part in a national screening program for the early detection of breast cancer over an eight-year period. Of the two patient subgroups (interval and screening), we found significant differences in the distribution of prognostic factors, with interval cases presenting at a lower mean age (p = 0.002), with higher percentages of human epidermal growth factor receptor 2 (HER-2) or triple negative and lower percentages of luminal A or luminal B carcinomas (p = 0.001), advanced stages (p<0.001), lower hormone receptor expression (p<0.001), poorer differentiation (p<0.001) and lower survival (p<0.001). Among the screening group, patients with tumors detected during the first screening round had a significantly lower mean age (p<0.001), a lower frequency of comorbidities (p = 0.038) and a lower tendency (p<0.1) to be diagnosed as triple negative breast carcinomas than incident cases. Our results highlight that breast tumors detected during the first screening round are frequently characterized by a more benign phenotype than the rest of the screening subgroups, which could be of help when stratifying the risk of death and selecting the best treatment option for each patient.

DNA methylation profiling identifies two distinct subgroups in breast cancers with low hormone receptor expression, mainly associated with HER2 amplification status

BackgroundCurrent clinical guidelines suggest that breast cancers with low hormone receptor expression (LowHR) in 1–10% of tumor cells should be regarded as hormone receptor positive. However, clinical data show that these patients have worse outcome compared to patients with hormone receptor expression above 10%. We performed DNA methylation profiling on 23 LowHR breast cancer specimens, including 13 samples with HER2 amplification and compared our results with a reference breast cancer cohort from The Cancer Genome Atlas to clarify the status for this infrequent but important patient subgroup.ResultsIn unsupervised clustering and dimensionality reduction, breast cancers with low hormone receptor expression that lacked HER2 amplification usually clustered with triple negative breast cancer (TNBC) reference samples (8/10; “LowHR TNBC-like”). In contrast, most specimens with low hormone receptor expression and HER2 amplification grouped with hormone receptor positive cancers (11/13; “LowHR HRpos-like”). We observed highly similar DNA methylation patterns of LowHR TNBC-like samples and true TNBCs. Furthermore, the Ki67 proliferation index of LowHR TNBC-like samples and clinical outcome parameters were more similar to TNBCs and differed from LowHR HRpos-like cases.ConclusionsWe here demonstrate that LowHR breast cancer comprises two epigenetically distinct groups. Our data strongly suggest that LowHR TNBC-like samples are molecularly, histologically and clinically closely related to TNBC, while LowHR HRpos-like specimens are closely related to hormone receptor positive tumors.

Outcome of breast cancer patients with low hormone receptor positivity: analysis of a 15-year population-based cohort

Guideline recommendations for the treatment of breast cancer with low hormone receptor (HR) expression (1%-9%) are ambiguous and several studies showed more similarities with HR-negative tumors than with HR strongly positive tumors (≥10%). We used a population-based 15-year cohort to compare patient characteristics and outcome of HR low positive tumors with HR-negative and HR strongly positive tumors, respectively. A total of 38 560 women diagnosed with early invasive breast cancer between 2004 and 2018 within the scope of the Munich Cancer Registry with 4.9 million inhabitants were included. Descriptive analyses of prognostic factors, treatment, and outcome analyses using the Kaplan-Meier method; cumulative incidence in consideration of competing risks; and multivariate analyses (Cox regression and Fine-Gray model) were conducted. Endpoints were time to local recurrence (TTLR), time to lymph node recurrence (TTLNR), time to metastasis (TTM), overall survival (OS), and relative survival (RS). A total of 861 patients (2%) had HR low positive, 4862 (13%) HR-negative, and 32 837 (85%) HR strongly positive tumors. Within the HER2-negative cohort (n= 33 366), survival of HR low positive tumors was significantly worse than that of HR strongly positive tumors [OS hazard ratio 0.66 (95% confidence interval 0.55-0.78)], whereas between HR low positive and HR-negative tumors no significant survival difference could be detected [OS hazard ratio 0.93 (95% confidence interval 0.78-1.11)]. TTLR, TTLNR, and TTM showed similar results. By contrast, within the HER2-positive cohort (n= 5194), no statistically significant differences between the three HR groups could be detected in multivariate analyses. Current definitions for HR positivity and its clinical relevance should be reconsidered. Patients with HR low positive/HER2-negative tumors could be regarded and treated similar to patients with triple-negative tumors.

Therapy response and prognosis of patients with early breast cancer with low positivity for hormone receptors – An analysis of 2765 patients from neoadjuvant clinical trials

AimTo evaluate HER2-negative breast cancer (BC) with a low hormone receptor (HR) expression, with regard to pathological complete response (pCR) and survival, in comparison to triple-negative BC (TNBC) and strong HR-positive BC. MethodsWe compared negative [oestrogen (ER) and progesterone receptor (PR) <1%], low-positive (ER and/or PR 1–9%) and strong-positive (ER or PR 10–100%) HR-expression in neoadjuvant clinical trial cohorts (n = 2765) of BC patients. End-points were disease-free survival (DFS), distant-disease free survival (DDFS) and overall survival (OS). We performed RNA sequencing on available tumour tissue samples from patients with low-HR expression (n = 38). ResultsNinety-four (3.4%) patients had low HR-positive tumours, 1769 (64.0%) had strong HR-positive tumours, and 902 (32.6%) had TNBC. There were no significant differences in pCR rates between women with low HR-positive tumours (27.7%) and women with TNBC (35.5%). DFS and DDFS were also not different [for DFS, hazard ratio 1.26, 95%-CI (confidence interval) : 0.87–1.83, log-rank test p = 0.951; for DDFS, hazard ratio 1.17, 95%-CI: 0.78–1.76, log-rank test p = 0.774]. Patients with strong HR-positive tumours had a significantly lower pCR rate (pCR 9.4%; odds ratio 0.38, 95%-CI: 0.23–0.63), but better DFS (hazard ratio 0.48, 95%-CI: 0.33–0.70) and DDFS (hazard ratio 0.49, 95%-CI: 0.33–0.74) than patients with low HR-positive tumours. Molecular subtyping (RNA sequencing) of low HR-positive tumours classified these predominantly into a basal subtype (86.8%). ConclusionLow HR-positive, HER2-negative tumours have a similar clinical behaviour to TNBC showing high pCR rates and poor survival and also a basal-like gene expression signature. Patients with low HR-positive tumours should be regarded as candidates for therapy strategies targeting TNBC.

N-myristoyltransferase proteins in breast cancer: prognostic relevance and validation as a new drug target

PurposeN-myristoyltransferases 1 and 2 (NMT1 and NMT2) catalyze the addition of 14-carbon fatty acids to the N-terminus of proteins. Myristoylation regulates numerous membrane-bound signal transduction pathways important in cancer biology and the pan-NMT inhibitor PCLX-001 is approaching clinical development as a cancer therapy. The tissue distribution, relative abundances, and prognostic value of the two human NMTs remain poorly understood.MethodsWe generated and validated mutually exclusive monoclonal antibodies (mAbs) specific to human NMT1 and NMT2. These mAbs were used to perform immunohistochemical analysis of the abundance and distribution of NMT1 and NMT2 in normal breast epithelial samples and a large cohort of primary breast adenocarcinomas from the BCIRG001 clinical trial (n = 706).ResultsNMT1 protein was readily quantified in normal and most transformed breast epithelial tissue and was associated with higher overall histologic grade, higher Ki67, and lower hormone receptor expression. While NMT2 protein was readily detected in normal breast epithelial tissue, it was undetectable in the majority of breast cancers. Detectable NMT2 protein correlated with significantly poorer overall survival (hazard ratio 1.36; P = 0.029) and worse biological features including younger age, higher histologic grade, lower hormone receptor expression, higher Ki67, and p53 positivity. Treatment of cultured breast cancer cells with PCLX-001 reduced cell viability in vitro. Daily oral administration of PCLX-001 to immunodeficient mice bearing human MDA-MB-231 breast cancer xenografts produced significant dose-dependent tumor growth inhibition in vivo.ConclusionsThese results support further evaluation of NMT immunohistochemistry for patient selection and clinical trials of NMT inhibition in breast cancer patients.

Abstract 1782: N-myristoyltransferase proteins in breast cancer: Prognostic relevance and validation as a new drug target

Abstract N-myristoyltransferases (NMTs) catalyze the addition of 14-carbon fatty acids to the N-terminus of proteins. This activity regulates numerous membrane-bound signal transduction pathways important in cancer biology, and the pan-NMT inhibitor PCLX-001 is in clinical development as a cancer therapy. The physiologic distribution and relative contributions of the two human NMTs, NMT1 and NMT2, remain poorly understood as previous studies used polyclonal antibodies with potential cross-reactivity. We generated and validated mutually exclusive monoclonal antibodies (mAbs) specific to the human isotypes of NMT1 and NMT2. These mAbs were used to perform an immunohistochemical (IHC) analysis of the abundance and distribution of NMT1 and NMT2 in normal human breast epithelial samples and a large (n=703) cohort of primary breast adenocarcinomas from the BCIRG001 study. While NMT1 protein was readily identifiable in most normal and transformed breast epithelial tissue, NMT1 abundance was associated with higher overall histologic grade, higher Ki67, and lower hormone expression. While NMT2 protein was readily detected in normal breast epithelial tissue, NMT2 protein was undetectable in the majority of malignant breast cancers, but detectable NMT2 protein correlated with significantly poorer overall survival outcomes (hazard ratio for death 1.36; p &amp;lt; 0.029) and significantly worse biological features including younger age, higher histologic grade, lower hormone receptor expression, higher Ki67, and p53 positivity. NMT status was unrelated to HER2 status. NMT1 and NMT2 protein abundances were positively correlated with each other. Treatment of cultured breast cancer cells with the pan-NMT inhibitor PCLX-001 reduced cell viability in vitro. Daily oral administration of PCLX-001 to immunodeficient mice bearing human MDA-MB-231 breast cancer xenografts induced significant dose-dependent tumor growth inhibition in vivo. These results support the further evaluation of NMT immunohistochemistry for selection of patients for NMT inhibitor therapy, and clinical trials of NMT inhibition in breast cancer patients. Citation Format: John R. Mackey, Justine Lai, Utkarsh Chauhan, Wei-Feng Dong, Darryl Glubrect, Sunita Ghosh, Gilbert Bigras, Raymond Lai, Luc G. Berthiaume. N-myristoyltransferase proteins in breast cancer: Prognostic relevance and validation as a new drug target [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 1782.

Does dose-dense neoadjuvant chemotherapy have clinically significant prognostic value in breast cancer?: A meta-analysis of 3,724 patients.

PurposeNeoadjuvant chemotherapy (NCT) is typically the initial treatment for non-early breast cancer patients. We thereby conducted a meta-analysis to explore whether dose-dense neoadjuvant chemotherapy (ddNCT) improved the long-term prognosis of patients compared to the standard NCT regimen.MethodsWe compared the differences in efficacy and prognosis between patients receiving standard NCT and ddNCT. We also calculated the pooled odds ratio (OR) of pathological complete response (pCR) and the pooled hazard ratio (HR) of overall survival (OS) and disease-free survival (DFS).ResultsNine randomized controlled trials involving 3,724 patients from 10 published studies were included in the meta-analysis. The pooled OR for ddNCT was 1.18 (95% confidence interval (CI): 0.83–1.67, P = 0.356). A subgroup analysis in the cases with low hormone receptor expression levels showed the pCR in patients undergoing ddNCT was significantly higher than the pCR in patients undergoing standard NCT (OR = 1.36, 95% CI: 1.09‒1.69, P = 0.007). There was no significant difference in DFS and OS between ddNCT and standard NCT (DFS: HR = 0.90, 95% CI: 0.79‒1.02, P = 0.095; OS; HR = 0.91, 95% CI: 0.81‒1.04, P = 0.160), regardless of hormone receptor expression levels. These data suggested the higher pCR rate in patients receiving ddNCT did not result in a survival benefit.ConclusionsThe meta-analysis demonstrated that ddNCT can significantly improve the pCR rate in patients with low hormone receptor expression levels, although patient survival was not significantly improved. The ddNCT can increase the breast-conserving rate and reduced pre-operative waiting time without increasing adverse reactions. This regimen can be considered when developing an NCT plan.

Low Positivity for Hormone Receptors in Early Breast Cancer: Therapy-Response and Prognosis -A Systematic Analysis of a Total of 9027 Patients

Low Positivity for Hormone Receptors in Early Breast Cancer: Therapy-Response and Prognosis -A Systematic Analysis of a Total of 9027 Patients

Abstract PD6-05: A prospective evaluation of clinical outcomes in women with pregnancy-associated breast cancer (PABC)

Abstract Background: Many studies suggest that women with PABC- breast cancer (BC) diagnosed during pregnancy or within 12 months post partum- have adverse outcomes compared to age matched women whose BC is not associated with pregnancy (non-PABC). However, it is unclear whether this is due to diagnostic delay alone or biological differences. Hence, we investigated whether PABC is independently prognostic for disease-free survival (DFS) and overall survival (OS) in a prospective database of young BC patients. Methods: A prospective database of women ≤40 years of age diagnosed with BC between February 2008 and January 2015 was analyzed. Data regarding, stage at diagnosis, pathology, treatment, and clinical outcomes were available. Statistically significant differences in baseline characteristics and administered therapies in women with and without PABC were evaluated using the chi-square or Fisher's Exact tests. Kaplan-Meier curves for DFS and OS in the PABC and non-PABC cohorts were compared using the log-rank test. A multivariate Cox proportional hazards model adjusted for age, nodal involvement and tumor size. Results: Of 224 women in the database who provided consent for research, 32 (12%) had PABC. Mean age of the PABC and non-PABC patients respectively was 34 (range 27 to 39) and 37 (range 21 to 40) and the median follow-up was 40 months in both groups. PABC was more likely to be locally advanced at diagnosis (44% vs. 22%, p&amp;lt;0.01) and less likely to be hormone receptor positive (75% versus 85%; p &amp;lt;0.01). There was no significant difference in age at diagnosis, tumor grade, lymphovascular invasion, HER2 expression or administered treatments between the two groups. Among the 166 women with early stage BC (not locally advanced), PABC was associated with positive lymph node status in a univariate model [OR 3.2 (95%CI 1.2-8.4), p=0.02] but just missed significance in a multivariate analysis that adjusted for age and tumor size (p=0.06). Eight patients (22%) in the PABC group and 19 (10%) in the non-PABC group experienced local or distant disease recurrence; 3 patients (8%) in the PABC group and 11 (6%) in the non-PABC group died. The 3-year DFS in the PABC and non-PABC cohorts was 79% vs. 90% (p=0.22) and the 3-year OS was 97% in both groups. Conclusion: Diagnostic delay could account for the higher rate of locally advanced disease in the PABC group. However, the lower hormone receptor expression and strong trend toward greater lymph node involvement independent of size suggest that women with PABC may have intrinsically worse disease biology. Event rates may still be too low to detect a statistically significant difference in recurrence risk. Further research is necessary to identify unique molecular features of PABC that may be amenable to targeting. Citation Format: Jerzak KJ, Zhu S, Li N, Mandel R, Warner E. A prospective evaluation of clinical outcomes in women with pregnancy-associated breast cancer (PABC) [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr PD6-05.

Hormone Receptor Expression Analyses in Neoplastic and Non-Neoplastic Canine Mammary Tissue by a Bead Based Multiplex Branched DNA Assay: A Gene Expression Study in Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Samples.

Immunohistochemistry (IHC) is currently considered the method of choice for steroid hormone receptor status evaluation in human breast cancer and, therefore, it is commonly utilized for assessing canine mammary tumors. In case of low hormone receptor expression, IHC is limited and thus is complemented by molecular analyses. In the present study, a multiplex bDNA assay was evaluated as a method for hormone receptor gene expression detection in canine mammary tissues. Estrogen receptor (ESR1), progesterone receptor (PGR), prolactin receptor (PRLR) and growth hormone receptor (GHR) gene expressions were evaluated in neoplastic and non-neoplastic canine mammary tissues. A set of 119 fresh frozen and 180 formalin-fixed, paraffin-embedded (FFPE) was comparatively analyzed and used for assay evaluation. Furthermore, a possible association between the hormone receptor expression in different histological subtypes of canine malignant mammary tumors and the castration status, breed and invasive growth of the tumor were analyzed. The multiplex bDNA assay proved to be more sensitive for fresh frozen specimens. Hormone receptor expression found was significantly decreased in malignant mammary tumors in comparison to non-neoplastic tissue and benign mammary tumors. Among the histological subtypes the lowest gene expression levels of ESR1, PGR and PRLR were found in solid, anaplastic and ductal carcinomas. In summary, the evaluation showed that the measurement of hormone receptors with the multiplex bDNA assay represents a practicable method for obtaining detailed quantitative information about gene expression in canine mammary tissue for future studies. Still, comparison with IHC or quantitative real-time PCR is needed for further validation of the present method.

Clinicopathological Features and Prognosis of Metaplastic Breast Carcinoma: Experience of a Major Chinese Cancer Center.

Metaplastic breast carcinoma (MBC) is a rare heterogeneous group of primary breast malignancies, with low hormone receptor expression and poor outcomes. To date, no prognostic markers for this tumor have been validated. The current study was undertaken to evaluate the clinicopathologic characteristics, the response to various therapeutic regimens and the prognosis of MBCs in a large cohort of patients from Tianjin Medical University Cancer Hospital in China. Ninety cases of MBCs diagnosed in our hospital between January 2000 and September 2014 were retrieved from the archives. In general, MBCs presented with larger size, a lower rate of lymph node metastasis, and demonstrated more frequent local recurrence/distant metastasis than 1,090 stage-matched cases of invasive carcinoma of no specific type (IDC-NST), independent of the status of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 expressions. The five-year disease-free survival (DFS) of MBC was significantly worse than IDC-NST. Using univariate analysis, lymph node metastasis, advanced clinical stage at diagnosis, high tumor proliferation rate assessed by Ki-67 labeling, and epidermal growth factor receptor (EGFR) overexpression/gene amplification were associated significantly with reduced DFS, while decreased OS was associated significantly with lymph node metastasis and EGFR overexpression/gene amplification. With multivariate analysis, lymph node status was an independent predictor for DFS, and lymph node status and EGFR overexpression/gene amplification were independent predictors for OS. Histologic subtyping and molecular subgrouping of MBCs were not significant factors in prognosis. We also found that MBCs were insensitive to neoadjuvant chemotherapy, routine chemotherapy, and radiation therapy. This study indicates that MBC is an aggressive type of breast cancer with poor prognosis, and that identification and optimization of an effective comprehensive therapeutic regimen is needed.

Abstract P3-14-20: Neoadjuvant chemotherapy in young age breast cancer: Survival benefit over adjuvant chemotherapy in clinically T2 node positive patients

Abstract Background: The downstaging of the primary tumor and the increase in breast conservation rates seems to be the only clinical benefit of Neoadjuvant systemic therapy(NST) in breast cancer treatment, given that several studies failed to demonstrate an improvement of overall survival compared with postoperative adjuvant chemotherapy. In Europe, S6 trial showed better early outcome in survival in favour of the neoadjuvant chemotherapy group compared to adjuvant chemotherapy group in premenopausal patients without significantly modifying long-term event rates. The aim of this study was to assess a potential advantage in survival by neoadjuvant as compared to adjuvant chemotherapy in young age breast cancer patients. Methods: Between January 2001 and December 2008, 1169 consecutive patients with breast cancer aged under 40 underwent adjuvant chemotherapy before or after surgery. Prospectively collected medical records for all patients were reviewed retrospectively. For the comparison of survival between neoadjuvant versus adjuvant chemotherapy group, cinically T2 and node positive patients were retrieved. Survival curves were derived from Kaplan-Meier estimates and compared by log-rank test. Results: Of the 1169 patients, 203(17.3%) patients were treated with neoadjuvant chemotherapy, and they were grouped as ‘NST’ and ‘non-NST’ according to initial treatment. About 47% patients in each group were clinically T2 patients. (99(47.8%) in NST group, 453(46.9%) in non-NST group) Among them, clinically T2 and node positive patients were 188, 97 patients in NST group, 91 patients in non-NST group each. The median age was 35.11±3.9 years old and HER2 amplification was observed as 23.5%, and they were not different between two groups.(p = 0.146 and 0.941 each) Significant lower hormone receptor expression rate and higher Ki-67 level were observed in NST group(p = 0.03 and &amp;lt;0.0001 each) Breast conservation surgery rate was also significantly different between two groups, more favorable results in NST group.(67% in NST group, 37.4% in non-NST group, p&amp;lt;0.0001) During median follow-up period of 61 months (range 44 to 148 months), we observed a statistically significant difference (p = 0.011) in survival in favour of the NST group. This benefit of survival was presented consistently regardless of hormone receptor expression. A similar trend was seen when the time to distant disease recurrence was evaluated (p = 0.176). And this trend was more prominent in hormone receptor negative patients, but still not statistically significant. (p = 0.144) The mean total dose of chemotherapy administered was similar in both groups. Improved survival figures in the NST group could be the result of the early initiation of systemic treatment, but the trend in favour of decreased metastases was not statistically significant. Conclusion: A potential advantage of primary over adjuvant chemotherapy in young age breast cancer patients’ survival might be proposed by this results. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-14-20.

Patients with nipple-areola Paget's disease and underlying invasive breast carcinoma have very poor survival: a matched cohort study.

Paget’s disease (PD) of the breast is a rare disease. The survival rate of PD was reported to depend on the characteristics of the underlying carcinoma. This study aimed to investigate the characteristics and survival rate of PD patients with underlying invasive breast carcinoma (IBC). Fifty-two patients were diagnosed with PD and an associated IBC from 2001 to 2005 in Fudan University Shanghai Cancer Center. Twenty-four (46.2%) had no clinical manifestation of PD and were diagnosed unexpectedly by a histologic examination. The 52 patients were all recruited in this study as the PD group. They tended to have greater chances of lymph node involvement (53.8% vs. 35.7%), lower hormone receptor expression (34.6% vs. 69.7%), higher human epidermal growth factor receptor 2 (HER2) expression (76.9% vs. 21.3%), and worse survival (5-year relapse-free survival (RFS) 52.2% vs. 86.7%, P<0.01; breast cancer-specific overall survival (OS) 62.1% vs. 91.8%, P<0.01) when compared with patients diagnosed with IBC. A matched study was then performed to investigate whether the poor survival of patients in the PD group was due to the unfavorable prognosis of the underlying IBC. One hundred and fifty-six (3∶1 ratio of controls to PD patients) patients diagnosed with IBC only were recruited into the matched group. The match was conducted according to four variables: dimension of IBC, lymph node status, hormone receptor status and HER2 status. The 5-year RFS (52.2% vs. 81.4%, P<0.01) and OS (62.1% vs. 85.9%, P<0.01) were both lower for patients in the PD group than those in the matched group. Patients with PD and underlying IBC had poor survival. Their survival was worse than that of patients with IBC of similar stage and characteristics. For patients with no clinical PD manifestation who were histologically diagnosed as PD, survival might be worse compared to patients with clinically diagnosed PD.

Abstract P6-07-42: Patients with Nipple-areola Paget's Disease and Underlying Invasive Breast Carcinoma had a Very Poor Survival: a Matched Cohort Study

Abstract Purpose: Nipple-areola Paget's disease together with invasive breast cancer is a rare kind of disease. This study is to investigate the characteristic and survival of it. Patients: From 2001 to 2005, all patients diagnosed as nipple-areola Paget's disease with underlying invasive breast carcinoma in Fudan University Shanghai Cancer Center (FUSCC) were recruited in this study (PD group). Patients diagnosed as invasive breast cancer without Paget's disease were recruited in control group for clinical and pathologic features comparison. Since tumors in PD group expressed more high risk factors (see results), a matched study was done to investigate whether the poor survival of PD group was by nature or due to the overexpression of high risk factors. The matched group was derived from all the patients who received mastectomy for invasive breast cancer (with no Paget's disease) during 2001 to 2005 in FUSCC. The match was conducted according to four variables: dimension of tumor, lymph node status, hormone receptor status and Her2 status. After matching, patients were randomly selected in a 3:1 ratio to patients in PD group. Results: 52 patients were recruited in PD group. 22 (42.3%) patients' primary symptom were nipple change, 25 (48.1%) patients' primary symptom were mass in breast. Physical examination found no nipple-areola change in 24 patients at diagnosis, their Paget's disease were found unexpectedly by routine nipple-areola section examination after mastectomy. More than 8000 invasive breast cancers were diagnosed during 2001 to 2005. 700 patients (140 consecutive patients per year, average monthly patient amount) were recruited in control group. Compared with control group, tumors in PD group had larger size, more axillary lymph node involvement, lower hormone receptor expression, higher HER2 expression and worse survival. 156 patients were recruited in the matched group. The 5-year relapse-free survival and overall survival were both lower for patients in PD group than those in the matched group (table1). Conclusions: Patients with invasive breast cancer and Paget's disease tended to have larger size, more lymph nodes involvement, lower hormone receptor expression, higher HER2 expression and worse survival than those without PD. The matched study confirmed the survival disadvantage of patients with Paget's disease and underlying invasive breast cancer. Thus, oncologists should regard such patients as having high risk for relapse. Pathologists should exam the nipple-areola section carefully after mastectomy although the patient has no macroscopic nipple-areola change. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-42.

Metaplastic breast carcinomas and their relationship with basal-like phenotype

Metaplastic carcinoma is a rare tumor showing high histological grade and low hormone receptor expression. It has pure epithelial and mixed types. Studies have suggested that metaplastic carcinomas may have a basal-like profile. Our aim was to evaluate the clinicopathological features of 11 metaplastic carcinomas and determine their resemblance to basal-like breast carcinomas regarding their morphological and immunohistochemical profile. Eleven metaplastic carcinoma cases were reviewed for their histopathological features. All tumors but one were evaluated for the immunohistochemical expressions of the cytokeratin 5/6, cytokeratin 14 and epidermal growth factor receptor; and hormonal status was assessed. Four of eleven cases were carcinoma with chondroid metaplasia, 3 were adenosquamous carcinoma, 2 were squamous cell carcinoma and 2 were carcinosarcoma. The mean patient age was 53 years and the mean tumor size was 5,1 cm. Histological grade was 3 for all with a nuclear grade of 3. Average mitotic count was 31/10 high power fields. Four cases had a central scar, 5 had central necrosis and 7 had geographic necrosis. Tumor growth pattern was pushing in 6 cases and no carcinoma in-situ was identified in 5 cases. Seven of 10 patients had axillary lymph node metastasis. Seven of 10 cases were triple-negative (estrogen receptor-, progesterone receptor-, HER2-) and 6 of them were positive for cytokeratin 5/6 and/or epidermal growth factor receptor, consistent with basal-like immunophenotype. Cytokeratin 14 was positive in 7 cases. Metaplastic carcinomas are large-sized, high-grade tumors with prominent nuclear pleomorphism and frequent mitosis. They rarely overexpress hormone receptors and HER2 and generally have basal-like immunophenotype.

The prognoses of metaplastic breast cancer patients compared to those of triple-negative breast cancer patients

Metaplastic breast carcinoma (MBC) is a rare, heterogeneous breast cancer characterized by admixture of adenocarcinoma with metaplastic elements, low hormone receptor expression, and poor outcomes. The authors retrospectively reviewed the medical records of 47 MBC patients and 1,346 invasive ductal carcinoma (IDC) patients. Two hundred eighteen of the IDC patients were triple-negative (TN-IDC) for estrogen receptor, progesterone receptor, and human epidermal growth factor receptor-2 (ER-/PR-/HER2-). Patients were surgically treated at the Samsung Medical Center between 2005 and 2009. The MBC patients presented with a larger tumor size, lower lymph node involvement, higher histological and nuclear grades, higher triple negativity (ER-/PR-/HER2-) and higher p53, CK5/6, and EGFR expressions compared with those of the IDC group. However, there were no significant differences in clinicopathological characteristics between MBC and TN-IDC. During the follow-up period (median duration of 30.3 months, range 2.6-56.3 months), seven (14.9%) MBC patients, and 98 (7.1%) IDC patients had disease recurrence. The three-year disease-free survival (DFS) rate was 78.1% in the MBC group and 91.1% in IDC group (P < 0.001). The three-year DFS rate was not significantly different between the MBC and TN-IDC groups (78.1 vs. 84.9%, P = 0.114). However, in patients with lymph node metastasis who underwent adjuvant chemotherapy, the three-year DFS rate was 44.4% in the MBC group and 72.5% in the TN-IDC group (P = 0.025). The authors found that MBC had a poorer clinical outcome than did IDC. In breast cancer patients with nodal metastasis, MBC had a poorer prognosis than did TN-IDC, despite adjuvant chemotherapy.

Abstract P3-10-25: The Prognosis of Metaplastic Breast Cancer Patients Compare to Triple-Negative Breast Cancer Patients

Abstract Background: Metaplastic breeast cancer (MBC) is a rare, heterogenous cancer characterized by admixture of adenocarcinoma with metaplastic elements, low hormone receptor expression and poor outcome. This study was planned to assess the clinicopathological chacteristics and immunohistochemical findings of MBC compared to invasive ductal carcinoma (IDC) including the triple-negative subtype (TN-IDC). Material and Methods: We retrospectively reviewed the medical records of 47 MBC and 1,346 IDC patients. Two hundred eighteen TN-IDC patients were included in the 1,346 IDC patients. Between 2005 and 2009, these patients were undergone surgical treatment at the Samsung Medical Center. Patients were reviewed clinicopathologic factors, immunohistochemistry of biologic factors such as ER, PR, HER-2, p53, Ki67, cytokeratine (CK) 5/6, epidermal growth factor receptor (EGFR), and treatment modalities (type of operation, use of chemotherapy, radiotherapy and hormone therapy). Result: The MBC patients presented with a larger tumor size (&amp;gt;T1, 66.0% vs. 44.3.%, P = 0.008), lower lymph node involvement (N0, 73.3% vs. 55.6%, P = 0.03), higher histologic (HG) and nuclear grade (NG) (HG3, 70.0% vs. 41.5%, P = 0.001; NG3,82.6% vs. 46.9%, P &amp;lt; 0.001), fewer estrogen receptor (ER), progesterone receptor (PR) and HER2 positivity (ER+, 4.3% vs. 69.2%, P &amp;lt; 0.001; PR+, 6.4% vs. 63.5%, P &amp;lt; 0.001; HER2+, 0% vs. 27.6%, P &amp;lt; 0.001), higher p53, CK5/6 and EGFR expression (p53+, 63.8% vs.38.8%, P &amp;lt; 0.001; CK5/6+, 71.9% vs.21.5%, P &amp;lt; 0.001; EGFR+, 93.9% vs.21.6%, P &amp;lt;0.001) and more TN subtypes (93.6% vs. 16.2%, P &amp;lt; 0.001) compared to the IDC group. There was no significant difference in clinicopathological characteristics with MBC and TN-IDC except EGFR over expression (EGFR+, 93.9% vs.69.0%, P = 0.017). In follow-up duration (median 30 months, range 2-56 months), seven (14.9%) MBC patients and 98 (7.2%) IDC patients recurred. The 3-year disease-free survival (DFS) rate was 78.1% in the MBC group and 91.1% in IDC group (P &amp;lt;0.001). The 3-year DFS rate was not significantly difference between MBC group and TN-IDC group (78.1% vs. 84.9%, P = 0.114). However, in patients with lymph node metastasis who underwent adjuvant chemotherapy, the 3-year DFS rate was 44.4% in MBC group and 72.5% in TN-IDC group (P = 0.025). Discussion: In our result, MBC show poorer clinical outcome than IDC. It is not shown significant difference between MBC and TN-IDC. However, MBC patients with nodal metastasis have poorer prognosis than TN-IDC patients with metastasis despite adjuvant chemotherapy. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-25.