Arisa Nishimukai

High Ki-67 Expression and Low Progesterone Receptor Expression Could Independently Lead to a Worse Prognosis for Postmenopausal Patients With Estrogen Receptor-Positive and HER2-Negative Breast Cancer

UNLABELLED We examined the prognostic significance of progesterone receptor (PgR) expression in immunohistochemical-based luminal subtypes defined by Ki-67 expression, taking menopausal status into consideration. The study included 327 surgically removed estrogen receptor-positive and human epidermal growth factor receptor 2-negative breast cancers. High Ki-67 expression (> 15%) and low PgR expression (£ 20%) were significant independent factors resulting in worse distant relapse-free survival. This association was observed in postmenopausalwomen but not in premenopausal women. BACKGROUND Accurate classification of luminal A and luminal B characteristics of estrogen receptor (ER)-positive and human epidermal growth factor receptor 2 (HER2)-negative breast cancer is considered clinically important for determining effective adjuvant treatment. Although Ki-67 expression has been identified as an essential constituent for this classification, the role of progesterone receptor (PgR) expression has yet to be fully elucidated. Because PgR expression is influenced by the estrogen milieu, we examined the prognostic significance of PgR expression in immunohistochemical (IHC)-based luminal subtypes defined by Ki-67 expression, taking menopausal status into consideration. MATERIALS AND METHODS We examined 327 surgically removed ER(+) and HER2(-) breast cancer specimens. ER, PgR, and Ki67 expression was determined IHC for semiquantitative measurement. We used 1%, 20%, and 15% as the cutoff value for ER, PgR, and Ki-67, respectively. RESULTS Breast cancer with low PgR (≤ 20%) expression was significantly associated with postmenopausal status, a large tumor size, and low ER expression. The low PgR expression subset had significantly worse distant relapse-free survival (DRFS) than the high PgR expression subset (P = .0067). This association was observed consistently in postmenopausal women but not in the premenopausal women. Multivariate analysis demonstrated that high Ki-67 expression (hazard ratio [HR], 3.80; 95% confidence interval [CI], 1.57-10.58; P = .003) and low PgR expression (HR, 2.54; 95% CI, 1.08-6.40; P = .038) were significant independent factors affecting DRFS. CONCLUSION Low PgR expression was independently associated with a poorer prognosis for ER(+) and HER2(-) breast cancer. Determination of PgR expression combined with that of Ki-67 could thus improve the accuracy of IHC-based classification of luminal A and luminal B breast cancer, especially for postmenopausal women.

Two Giant Renal Aneurysms and Renal Arteriovenous Fistula Associated with Cardiac Insufficiency and a Sustained Elevation of Atrial Natriuretic Peptide and Brain Natriuretic Peptide

A 64-year-old man presented with chief complaints of exertional dyspnea and palpitation. He had previously undergone left nephrolithotomies twice. A chest roentgenogram showed pleural effusion on both sides with cardiac dilation, and electrocardiography showed a frequent occurrence of ventricular premature contractions. An echocardiogram showed diffuse hypokinesis of the left ventricular wall motion (ejection fraction, 45%) and dilation of the left ventricle (left ventricular end-diastolic dimension, 61 mm). We administered diuretics, ACE inhibitors and a β-adrenergic blocking agent after making a diagnosis of cardiac insufficiency. Because coronary angiography showed 90% stenosis of the left anterior descending coronary artery (No. 7), we performed coronary angioplasty in this locus. Though both the left ventricular wall motion and ejection fraction improved, and the clinical symptoms disappeared, the left ventricular end-diastolic dimension, and arrhythmia did not improve. Furthermore, the brain natriuretic peptide increased despite these treatments. Thereafter, a left renal artery aneurysm (extrarenal aneurysm measuring 5 cm in diameter and an intrarenal aneurysm measuring 3 cm in diameter) and a left renal arteriovenous fistula were discovered when abdominal echography was performed because of epigastric discomfort. As a result, a left total nephrectomy was performed. Subsequently, the left ventricular end-diastolic dimension and arrhythmia improved, and the brain natriuretic peptide returned to a normal value. We herein report a case that developed cardiac insufficiency due to a renal aneurysm and renal arteriovenous fistula after undergoing left nephrolithotomies twice.

Activation of mTOR/S6K But Not MAPK Pathways Might Be Associated With High Ki-67, ER+, and HER2− Breast Cancer

UNLABELLED We determined the activation of the phosphoinositide 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling pathways in 108 cases of estrogen receptor-positive and human epidermal growth factor receptor 2-negative breast cancer with high and low Ki-67 expression. The expression levels of Ki-67, p53, phosphorylated MAPK (pMAPK), and protein S6 (pS6; downstream molecule of PI3K/Akt/mammalian target of rapamycin/S6 kinase pathway) were determined immunohistochemically. pS6 positivity, but not pMAPK positivity, was significantly associated with the high Ki-67 expression subset. BACKGROUND Evaluation of luminal A and luminal B characteristics of estrogen receptor (ER)-positive and human epidermal growth factor receptor 2 (HER2)-negative breast cancer is considered important. Although the phosphoinositide 3 kinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling pathways are thought to be involved in the luminal B subtype, the details of their contribution to breast cancer remain unclear. MATERIALS AND METHODS We determined the activation of these pathways (phosphorylated MAPK [pMAPK] and protein S6 [pS6; a downstream molecule of PI3K/Akt/mammalian target of rapamycin (mTOR)/S6 kinase (S6K)]) in 108 ER(+), HER2(-) breast cancer cases with high and low Ki-67 expression. The ER, progesterone receptor (PgR), Ki-67, p53 expression levels were also determined immunohistochemically. The cutoff value for Ki-67 was set at 15%. RESULTS A significantly greater percentage of cancer cases with high Ki-67 expression showed pS6 positivity than did those with low Ki-67 expression (53.2% vs. 19.7%; P = .0003). No significant differences were found between the cases with high and low expression levels were detected for p53 (23.4% vs. 11.5%; P = .12) or pMAPK (36.2% vs. 34.4%; P = .85) positivity. Multivariate analysis showed that pS6 positivity (odds ratio 5.16, 95% confidence interval 1.95-13.63; P = .0009), nuclear grade 2 and 3, and low PgR expression (≤ 20%) were independently associated with the high Ki-67 subset. CONCLUSION From our findings, we have concluded that the pS6 expression level is associated with the characteristics of breast cancer with high Ki-67 expression. Because these associations were observed, irrespective of menopausal status, the biologic difference seems to be less affected by estrogen signaling than by activation of S6 protein, especially in terms of proliferation. Our findings have also indicated that targeting the mTOR/S6K pathway might be a useful strategy for the treatment of ER(+)/HER2(-) breast cancer with high Ki-67 expression.

High levels of serum CA15-3 and residual invasive tumor size are associated with poor prognosis for breast cancer patients with non-pathological complete response after neoadjuvant chemotherapy: FUJIMOTO et al.

To identify surrogate markers for prognosis of breast cancer patients with non‐pathological complete response (non‐pCR) to neoadjuvant chemotherapy (NAC), our investigation focused on the serum levels of carcinoembryonic antigen (CEA) and carbohydrate antigen (CA15‐3) as well as clinicopathological factors both before and after NAC.

Significant Association Between Low Baseline Neutrophil-to-Lymphocyte Ratio and Improved Progression-free Survival of Patients With Locally Advanced or Metastatic Breast Cancer Treated With Eribulin But Not With Nab-Paclitaxel

Introduction Although eribulin and nab‐paclitaxel are chemotherapy agents widely used for locally advanced or metastatic breast cancer (MBC), their predictive factors remain unknown. Because the absolute neutrophil‐to‐lymphocyte ratio (NLR) is a significant prognostic factor for early‐stage breast cancer, we investigated its usefulness in terms of the eribulin or nab‐paclitaxel treatment efficacy for MBC. Patients and Methods A total of 85 patients with MBC treated with eribulin (n = 59) or nab‐paclitaxel (n = 26) were recruited. NLR values were collected at baseline, after 1 cycle, after 2 cycles, and at the end of treatment. The NLR cutoff value was set at 3. Results The progression‐free survival (PFS) of patients with an NLR < 3 at baseline (median, 242 days; n = 24) was significantly better than that of patients with an NLR of ≥ 3 (median, 98 days; n = 35; hazard ratio, 0.37, 95% confidence interval, 0.18‐0.71; P = .0032). Similarly, the overall survival was marginally significantly better in patients with an NLR < 3 who were treated with eribulin (P = .058). However, the NLR was not significantly associated with PFS or overall survival for patients treated with nab‐paclitaxel. No significant association was found between the NLR during treatment and PFS in the eribulin group. The significance of the NLR for the efficacy of eribulin was consistent, irrespective of estrogen receptor status, previous anthracycline or endocrine use, and the number of previous chemotherapy regimens. Conclusion A low NLR at baseline was significantly associated with improved PFS in patients treated with eribulin but not in those treated with nab‐paclitaxel. Therefore, the baseline NLR might be clinically useful for selecting patients who would benefit from eribulin. Micro‐Abstract Both eribulin and nab‐paclitaxel are widely used and effective chemotherapy agents for metastatic breast cancer; however, their predictive factors remain unknown. The usefulness of the neutrophil‐to‐lymphocyte ratio (NLR) in terms of treatment efficacy was investigated. We observed that a low NLR at baseline might be a significant indicator of improved outcomes for patients treated with eribulin but not with nab‐paclitaxel.

Tumor size and proliferative marker geminin rather than Ki67 expression levels significantly associated with maximum uptake of 18F-deoxyglucose levels on positron emission tomography for breast cancers

It has been well established that maximum standardized uptake value (SUVmax) for 18F-fluorodeoxyglucose positron-emission tomography/computed tomography (FDG PET/CT) is clinically useful for evaluating treatment efficacy as well as predicting prognosis of breast cancer patients. Although SUVmax reflects increased glucose uptake and metabolism possibly induced by activation of growth factor signaling or TP53 dysfunction, tumor characteristics of SUVmax-high breast cancers remain to be elucidated. For the present study, we used immunohistochemical staining to investigate expressions of phospho-ribosomal protein S6 (pS6, downstream molecule of phosphatidyl inositol 3-kinase/Akt/mammalian target of the rapamycin/S6K pathway) and phosphor-p44/42 mitogen-activated protein kinase (pMAPK). Expression levels of TP53 and proliferative marker geminin as well as Ki67 were also examined by means of immunostaining in 163 invasive breast cancers. Cutoff values were set at 10% for pS6, 20% for pMAPK and TP53, and 4% for geminin. The SUVmax levels were significantly higher in the pS6-positive (p = 0.0173), TP53-positive (p = 0.0207) and geminin-high cancers (p<0.0001), but there was no significant association between pMAPK expression levels and SUVmax levels. Multivariable analysis showed that a high geminin level (odds ratio: 6.497, 95% confidence interval: 2.427–19.202, p = 0.0001) and large tumor size (6.438, 2.224–20.946, p = 0.0005) were significantly and independently associated with SUVmax-high. Univariable but not multivariable analysis indicated that Ki67-high significantly correlated with SUVmax-high. Twenty of 23 (87.0%) breast cancers with tumor size >2cm and geminin-high showed SUVmax-high, while only 6 of 49 (12.2%) breast cancers ≤2cm in size and with low geminin levels were SUVmax-high. In conclusion, we could determine that breast cancers with a large tumor and a geminin-high rather than Ki67-high proliferative marker were significantly associated with high levels of SUVmax. These findings may signify that SUVmax reflects tumor characteristics with high proliferative activity but not activation of mTOR/S6K and MAPK pathways or increased glucose metabolism due to dysfunction of TP53.

Abstract P2-05-27: Baseline serum CA15-3 levels are associated with prognosis for breast cancer patients with non-complete pathological response to neoadjuvant chemotherapy

Background: It has been well demonstrated that patients who achieved pathological complete response (pCR) after neoadjuvant chemotherapy (NAC) had a favorable prognosis compared with patients who did not (non-pCR). Even though pCR was not attained, reduction in tumor volume after chemotherapy may be associated with improved prognosis for a certain number of patients. However, the association between residual tumor volume and prognosis is not necessarily consistent. In order to identify substitute markers for breast cancer patients with non-pCR after NAC, we investigated the impact of serum levels of carcinoembryonic antigen (CEA) and carbohydrate antigen (CA15-3) at baseline as well as post-NAC. Patients and Methods: Ninety-six breast cancer patients treated with NAC and operated on at the Hyogo College of Medicine were recruited for this study. Serum CEA and CA15-3 were measured prior to chemotherapy as well as at completion of pre-operative treatment. The optimal cutoff points for CEA (1.55ng/m, normal range: Results: pCR and non-pCR was attained by 21 and 75 patients, respectively. For the non-pCR patients, serum CEA levels at baseline were classified into high (n=35) and low (n=38) and serum CA15-3 levels at baseline into high (n=31) and low (n=43). RFS of non-pCR patients with high serum CA15-3 levels was significantly worse than of those with low levels (3-year RFS: 0.47 vs 0.93; p=0.0009). RFS for patients with high and low serum levels of CA15-3 after NAC was also significantly different (p=0.037). As for CEA, no significant association with RFS was observed either at baseline or post-NAC. Univariate analysis demonstrated that tumor size and baseline CA15-3 were significant prognostic factors for RFS. Multivariate analysis showed that both tumor size (hazard ratio (HR): 3.88, 95% confidence interval (CI): 1.21-12.35, p=0.023) and baseline CA15-3 (HR: 13.51, 95% CI: 1.74-105.08, p=0.013) were significant and independent risk factors for relapse. As for lymph node metastasis, tumor grade, residual tumor size and pre- and post-NAC Ki67 expression levels of patients with non-pCR showed no significant association with RFS. Conclusion and discussion: High levels of serum CA15-3 at baseline constituted a significantly worse prognosis for breast cancer patients with non-pCR. Tumor size at baseline but not residual size and baseline CA15-3 seems to suitable as a substitute for prediction of outcome for patients with non-pCR. Our findings suggest that these markers may be useful for identifying patients with poor prognosis who may be candidates for additional adjuvant treatment. Citation Format: Fujimoto Y, Imamura M, Higuchi T, Nishimukai A, Yanai A, Miyagawa Y, Murase K, Takatsuka Y, Miyoshi Y. Baseline serum CA15-3 levels are associated with prognosis for breast cancer patients with non-complete pathological response to neoadjuvant chemotherapy [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 P2-05-27.

Abstract P6-16-01: Differences in patterns of change of bone turnover markers during treatment with bone-modifying agents of breast cancer patients with bone metastases

Background: Bone-modifying agents have demonstrated their efficacy for treatment by suppressing osteoclast function. The activity of bone-modifying agents can be monitored by means of bone resorption markers such as c-terminal crosslinking telopeptide of type I collagen (1CTP) and N-telopeptide of type I collagen (NTX) as well as bone forming marker bone-specific alkaline phosphatase (BAP). In contrast to these markers which indirectly indicate bone turnover, tartrate-resistant acid phosphatase-5b (Tracp-5b) has been established as a direct marker showing osteoclast number and activity. The aim of this study was to identify the relative significance of these bone turnover markers as indicators of treatment efficacy induced by bone-modifying agents for breast cancer patients with bone metastases. Patients and Methods: For this study, 52 breast cancer patients with bone metastases treated with bone-modifying agents were recruited. Zoledronic acid and denosumab were administered as bone-modifying agents to 36 and 22 patients, respectively (for 6 patients, denosumab was used after zoledronic acid). Serum Tracp-5b, 1CTP, NTX and BAP were measured with, respectively, the EIA (enzyme immunoassay), RIA (two-antibody radioimmunoassay), ELISA (enzyme-linked immunosorbent assay) and CLEIA (chemiluminescent enzyme immunoassay) method. Blood samples were obtained pretreatment and 1, 3 and 6 months after treatment. Changes in these bone turnover markers were statistically analyzed with Friedman9s test, and correlation between serum markers and clinicopathological factors was calculated with Mann-Whitney9s test. Results: Serum tracp-5b decreased significantly after treatment (p Conclusion and discussion: Although baseline values of the bone turnover markers Tracp-5b, NTX and BAP decreased significantly after treatment with bone-modifying agents, the pattern of reduction for these three markers varied. Tracp-5b appears to reflect efficacy of bone-modifying agents most quickly and sensitively, possibly due to its direct link to the number and activity of osteoclasts. These findings may prove usefulness of Tracp-5b when considering the efficacy of various bone-modifying agents in clinical practice. Citation Format: Higuchi T, Nishimukai A, Yanai A, Miyagawa Y, Murase K, Imamura M, Ozawa H, Takatsuka Y, Miyoshi Y. Differences in patterns of change of bone turnover markers during treatment with bone-modifying agents of breast cancer patients with bone metastases. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P6-16-01.

Abstract P5-09-05: Prediction of bone metastases of breast cancer using combined markers of bone metabolism and inflammation

Introduction Bone metastases in breast cancer impair a patient9s QOL because of skeletal-related events such as bone pain, fractures, spinal cord compression, and hypercalcemia. It might be important to predict bone metastases and initiate adequate treatment early in the disease process. Screening and diagnosis of bone metastases are performed using serum markers and imaging systems such as CT, MRI, PET, and SPECT in postoperative testing. However, a method for predicting bone metastases for stratifying patients who require treatment has not been established. Although various markers of bone metabolism have been approved for monitoring of postoperative bone metastases, these are not considered clinically practical because of their low specificity. We selected TRACP-5b as a marker of bone metabolism; likelihood of bone metastases, and CRP as a marker of inflammation; likelihood of distant recurrence. We hypothesized that the combination of these two markers of different aspects would provide an accurate prediction of bone recurrence. Patients and methods Three hundred forty-nine breast cancer patients who underwent surgery in our hospital between August 5, 2010, and October 31, 2013, were enrolled in this study. Their serum levels of TRACP-5b and CRP were measured in a blinded manner at the R & D laboratory of Nittobo Medical Co., Ltd. Eighty-one patients were excluded (78 cases; neoadjubant chemotherapy administration, 3 cases; T4), and the data from the remaining 268 patients were included in the statistical analysis. The cutoff values were 380mU/dL for TRACP-5b and 0.016 mg/dL for CRP. Patients with both values above the cutoff value were classified as +/+, and they were compared with the other patients. The odds ratio between +/+ and the others were calculated using MedCalc statistical software. Results Patients stratified into four classes according to the value of TRACP-5b and CRP: +/+ (n=60), +/- (n=49), -/+ (n=76) and -/- (n=83), (+ means above the cutoff value). Eight of the 268 patients had relapsed metastases: three in the bone only, one in the bone and lung, three in lymph nodes only, and one in the lung only). The Incidence of bone metastases was 5 %(3/60) in the +/+ patients and 0.5 %(1/208) in the others. The incidence was significantly higher in the +/+ patients than in the others(odds ratio: 10.9, 95% CI 1.11 to 106.74, p= 0.040). When the other relapses not including bone metastases were included in the analysis, no significant difference was observed between the two groups (odds ratio: 0.4, 95% CI 0.02 to 7.07, P=0.513). TRACP-5b concentration alone could not classify the patients into two groups according to significantly different incidences of bone metastases(odds ratio: 4.5, 95% CI 0.46 to 43.57, P=0.197). Conclusion The results presented here show that the prediction of bone metastases by the combination of TRACP-5b and CRP concentrations is clinically relevant. We plan to increase the number of patients to provide sufficient statistical power to confirm this diagnostic potential. Citation Format: Arisa Nishimukai, Naoya Shibata, Wataru Kikuchi, Hiroki Hutawatari, Hideki Ishihara, Yasuo Miyoshi. Prediction of bone metastases of breast cancer using combined markers of bone metabolism and inflammation [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P5-09-05.