Makiko Kamio

Optimization of Region of Interest Drawing for Quantitative Analysis: Differentiation Between Benign and Malignant Breast Lesions on Contrast-Enhanced Sonography.

This study was performed to evaluate the diagnostic utility of quantitative analysis of benign and malignant breast lesions using contrast‐enhanced sonography.

Long-Term Follow-Up of Node-Negative Breast Cancer Patients Evaluated via Sentinel Node Biopsy After Neoadjuvant Chemotherapy

Micro‐Abstract The purpose of this study was to assess the usefulness of sentinel node biopsy (SNB) after neoadjuvant chemotherapy (NAC) in patients with clinically node‐negative breast cancer. SNB after NAC was as accurate as SNB without NAC. Axillary recurrence‐free survival rates were excellent regardless of whether NAC was performed before SNB. Background: Sentinel node biopsy (SNB) is used to accurately assess axillary lymph node status in patients with node‐negative breast cancer. However, its use after neoadjuvant chemotherapy (NAC) is controversial. We retrospectively assessed the usefulness of SNB after NAC by comparing axillary recurrence rates and other parameters in patients with clinically node‐negative breast cancer who underwent SNB after NAC or without NAC. Patients and Methods: At our hospital, 1179 patients with clinically node‐negative breast cancer underwent SNB from April 2007 to December 2013. The clinicopathological and survival data of patients who underwent SNB after NAC (the NAC group) and those who underwent SNB without NAC (the control group) were compared. Patients with a metastatic sentinel node underwent axillary lymph node dissection. Results: The number of patients in the NAC and control groups was 183 (15.5%) and 996 (84.5%), respectively. At diagnosis, tumors were significantly larger in the NAC group (P < .0001). Sentinel nodes were identified in almost all patients in both groups (99.5% in the NAC group vs. 99.8% in the control group). They were nonmetastatic in 147 (80.8%) patients in the NAC group and 849 (85.5%) patients in the control group. At the median follow‐up time of 51.1 months, 6 patients (0.6%) in the control group had axillary lymph node recurrence compared with no patients in the NAC group. Conclusion: SNB after NAC was as accurate as SNB without NAC in patients with clinically node‐negative breast cancer. Axillary recurrence‐free survival rates were excellent regardless of whether NAC was performed before SNB.

An extracellular matrix molecule, secreted by the epithelial-mesenchymal transition is associated with lymph node metastasis of thyroid papillary carcinoma.

Background: Papillary thyroid carcinoma often has lymph node metastasis, compared with follicular thyroid carcinoma. The study showed that epithelial-mesenchymal transition occurs in carcinoma cells during the first stage of metastasis, where some extracellular matrix molecules are secreted in large quantities. Sialic acid carried by fibronectin as the antigen of the monoclonal antibody (MoAb) JT-95, was detected in 90% of papillary thyroid carcinoma cases, and in a few follicular thyroid carcinomas, in the extracellular matrix of thyroid carcinoma cells. Objectives: The current study was conducted to investigate the association between increasing the number of extracellular matrix molecules, fibronectin, and lymph node metastasis. We also co-cultured a thyroid carcinoma cell line and lymphocyte cell line, with and without MoAb JT-95, in order to investigate the mechanism of cell to cell interaction. Patients and Methods: Immunostaining with JT-95 was performed in 45 papillary thyroid carcinoma cases, and 20 follicular type tumors, to investigate the association between the quantity of fibronectin expression and the frequency of lymph node metastasis. The thyroid carcinoma cell line (SW1736), which secreted fibronectin, and the B cell-lymphoma cell line (Daudi), which held integrin on the cell surface, were co-cultured to observe the adhesion of cells to each other. The SW1736 cell line, pretreated with JT-95, was also co-cultured with the Daudi cell line. Results: There were 39 cases with lymph node metastasis in 59 malignant tumors, and 0 cases in 6 benign follicular type tumors. The staining scores by JT-95 of the 39 tumors with lymph node metastasis were 5+ in eight cases and 6+ in 31 cases. On the other hand, the scores of 20 malignant tumors without lymph node metastasis were < 4+ in all of the cases. In the co-cultured assay, numerous adhesions were observed between the SW1736 and Daudi cells. In contrast, the inhibition of adherences was observed in proportion to the concentrations of JT-95. Conclusions: Increased fibronectin expression in thyroid malignancies is correlated with lymph node metastasis.

Abstract P5-07-07: DYRK2 contributes to the generation of breast cancer stem cells through KLF4

Cancer stem cells (CSCs) have been defined by the potential to self-renew and to differentiate. CSCs pose a major hurdle in the treatment of cancer. However, the mechanisms by which cells acquire CSC properties such as drug resistance remain unclear. Dual-specificity tyrosine-regulated kinase 2 (DYRK2) is a protein kinase that phosphorylates its substrates on serine/threonine. Initially, we found that DYRK2 phosphorylates p53 at Ser 46 to regulate apoptotic cell death in response to DNA damage. Recently, we have shown that DYRK2 controls Snail degradation in breast cancer and ovarian serous adenocarcinoma. We also found that knockdown of DYRK2 in luminal-type breast cancer MCF-7 cells increased the cancer stem cell population. Kruppel-like factor 4 (KLF4) is one of the Yamanaka factors. It has been reported that pluripotent stem cells from mouse embryonic or adult fibroblasts are induced by introducing four factors, Oct3/4, Sox2, c-Myc, and Klf4. This finding led us to determine if KLF4 is indispensable for the maintenance of CSCs. The aim of this study is to clarify whether DYRK2 regulates CSCs through KLF4 in breast cancer. Cell lines: MCF-7 (human mammary carcinoma: ATCC) cells were grown according to standard protocols. We established stable DYRK2-depleted cells. MCF-7 cells were transfected with pSuper-puro vector (pSuper control) or pSuper-puro DYRK2 shRNAs (shDYRK2) with puromycin to isolate stable cell lines. In turn, we established both stable DYRK2- and KLF4-depleted cells. shDYRK2 cells were transfected with pSuper-neo vector (pSuper-neo control) or pSuper-neo KLF4 shRNAs (shKLF4) with puromycin and G418. Knockdown of DYRK2 or KLF4 was confirmed by real-time RT-PCR and immunoblotting. The depleted cells were compared with the control cells using real-time RT-PCR, immunoblotting, flow cytometric analysis, mammosphere assay, xenograft models and immunohistological staining. We analyzed the population of breast cancer stem cells by flow cytometric analysis and in vitro mammosphere assay. The results showed that knockdown of DYRK2 was associated with the increase of CD44+/CD24- cells. While pSuper control cells formed mammospheres, they did in a lesser extent compared to shDYRK2 cells. In real-time RT-PCR and immunoblotting analysis, stable DYRK2 depletion in MCF-7 cells induced KLF4 accumulation. We then investigated the effect of KLF4 on stemness by flow cytometric analysis and in vitro mammosphere assay. The results showed that knockdown of KLF4 in shDYRK2 cells reduced the proportion of CD44+/CD24- cells. Whereas shDYRK2/shKLF4 cells formed mammospheres, they did in a lesser extent compared to shDYRK2/pSuper-neo control cells. Moreover, the scale of the mammospheres formed in shDYRK2/shKLF4 cells was significantly smaller, as compared with that in shDYRK2/pSuper-neo control cells. In xenograft models, the loss of KLF4 protein expression significantly decreased tumor formation. Immunohistological staining of fifty-nine samples from surgically treated breast cancer patients showed an inverse correlation between DYRK2 and KLF4 expression. These findings revealed that DYRK2 contributes to the generation of breast cancer stem cells through KLF4. Citation Format: Imawari Y, Mimoto RK, Yamaguchi N, Kamio M, Kato K, Nogi H, Toriumi Y, Uchida K, Takeyama H, Yoshida K. DYRK2 contributes to the generation of breast cancer stem cells through KLF4 [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 P5-07-07.

Nonrecurrent inferior laryngeal nerves and anatomical findings during thyroid surgery: report of three cases

A nonrecurrent inferior laryngeal nerve (NRILN) is found more frequently on the right side than on the left, and it is closely associated with an aberrant right subclavian artery. The presence of the aberrant right subclavian artery on preoperative computed tomography (CT) scan suggests NRILN; however, different types of branching locations and pathways exist. Here, we report three NRILN cases with different pathways where the vagus nerve arises more medial than usual and a review of the literature. Case 1: A 30-year-old Japanese female presented with papillary thyroid carcinoma. Preoperative CT scan revealed an aberrant right subclavian artery, and an operation was performed under suspicion of NRILN. During the operation, the vagus nerve was found to arise more medially than usual and two NRILNs originated from it at the level of the cricoid cartilage and at a more caudal position; the two NRILNs were preserved. Case 2: A 33-year-old Japanese female with a thyroid nodule of increased size underwent surgery. Preoperative CT scan revealed an aberrant right subclavian artery, which suggested NRILN. During the operation, the vagus nerve was identified to run more medially than usual and NRILN was found to originate at the level of the cricoid cartilage; NRILN was preserved. Case 3: A 78-year-old Japanese female underwent an operation with a diagnosis of papillary thyroid carcinoma. Preoperative CT scan showed an aberrant right subclavian artery. During the operation, NRILN was found to originate from the vagus nerve at the level of the lower pole of the thyroid gland, and the vagus nerve ran medial to the common carotid artery at the caudal level.

Abstract P2-12-14: The effect of low intensity exercise for mental status and quality of life of Japanese early breast cancer patients

Objective: Breast cancer is one of the most common cancers, and it accounts for 20% of malignant tumors of women. Psychological damage of the breast cancer patient is serious. In our institution, approximately 30% of the early breast cancer patients diagnosed as depressed mental status before operation. Breast cancer patients have to receive hormone therapy, chemotherapy, and radiation therapy for a long term after operation. Depressed mental status affects execution of the treatment, and the interruption of the treatment brings the risk of cancer recurrence. As a treatment for depressed mental status of cancer patients, medication and psychotherapy are common, but some previous reports showed exercise is effective. Exercises provided in these reports were various kinds and strength, and the standard prescription is not established. We investigate the effect of walking program as a mild aerobic exercise, whether it improves the depressed mental status and quality of life (QOL) of breast cancer patients. Methods: 25 early breast cancer patients were recruited. Depressed mental status and QOL were assessed by Center for Epidemiologic Studies Depression Scale (CES-D) and Medical Outcomes Study 36-item Short Form Health Survey (SF-36). Physical activity was measured by accelerometer (Lifecorder PLUS, Suzuken), and estimated using metabolic equivalent foe task (MET). Participants mounted accelerometer after discharge, and baseline physical activity was recorded till 1 month after operation. After recording baseline physical activity, we instructed to perform walking or mild aerobic exercise. Scores of CES-D and SF-36, and physical activity data were measured again after two months. Results: Physical activity after intervention was significantly increased as compared to baseline (8.5±5.5 vs 12.3±6.9 MET.h/week, p=0.02 ). At pre-operation phase, nine patients (36%) regarded as depressed mental status by CES-D, and depressed patients decreased to five at the time of 3 months after operation. A subscale of SF-36 ( mental health ) reduced significantly compared with pre-operation and 3 months after operation ( p Conclusion: This study demonstrated the possibility that walking program as a mild exercise improves mental status and QOL of perioperative breast cancer patients. It is necessary to examine in more detail for clinical application. Citation Format: Makiko Kamio, Kazumi Kawase, Hiroko Nogi, Ken Uchida, Hiroshi Takeyama. The effect of low intensity exercise for mental status and quality of life of Japanese early breast cancer patients [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 P2-12-14.

Abstract P2-06-14: Clinical relevance and biological properties of oligometastatic breast cancer in lung; prognostic impact of CD44+/CD24−/low cells

Background: Metastatic breast cancer is a systemic disease. Our aim was to evaluate the clinical outcomes of pulmonary metastasectomy of recurrent breast tumors and to identify possible prognostic factors. Methods: We reviewed data from a registry of patients with lung metastases from breast tumors who received pulmonary metastasectomy in Jikei University Hospital between 2004 and 2011. We analyzed prognostic factors for overall survival (OS) and progression free survival (PFS) after metastasectomy. We also investigated lung metastases for the prevalence of CD44+/CD24−/low tumor cells and evaluated their prognostic significance. Results: Among 17 patients with lung metastasis of breast tumors, 5-year OS and PFS were 72% and 36%, respectively. Better OS was observed among patients with oligometastatic breast cancer (OMBC). Patients with OMBC, estrogen receptor (ER) positive cells, and disease free intervals (DFI) of >8 years had better PFS. The average prevalence of CD44+/CD24−/low tumor cells in lung metastases of breast cancer was 21%, ranging from 0 to 90%. The presence of CD44+/CD24−/low tumor cells influenced the progression after lung metastasectomy, with median PFS times of only 6 months in patients with high-prevalence of cancer-initiating cells. CD44+/CD24−/low cells with cancer-initiating properties were present in only 9% ± 12 of patients with OMBC but were found in 73% ± 21 of patients with non-OMBC. Conclusion: Pulmonary metastasectomy may be a treatment option for OMBC patients with lung metastases. Better prognosis of OMBC may be related to low levels of cancer-initiating cells. Citation Format: Rei Mimoto, Tadashi Kobayashi, Yoshimi Imawari, Makiko Kamio, Kumiko Kato, Hiroko Nogi, Yasuo Toriumi, Ken Uchida, Hiroshi Takeyama. Clinical relevance and biological properties of oligometastatic breast cancer in lung; prognostic impact of CD44+/CD24−/low cells [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 P2-06-14.

Abstract P2-01-19: Long-term follow-up of the node-negative breast cancer patients before treatment evaluated with sentinel node biopsy alone following neoadjuvant chemotherapy

BACKGROUND Sentinel node biopsy (SNB) for the node negative breast cancer is standard treatment as an accurate assessment of axillary lymph node status. The use of neoadjuvant chemotherapy (NAC) has increased during the past several years. At present, the use of SNB following NAC is controversial. Proponents of SNB after NAC prefer a single surgical procedure with potential for fewer axillary dissections. OBJECTIVE Our objective was to examine SNB evaluation alone following NAC in patients with clinically node-negative breast cancer before treatment and to evaluate the axillary lymph node recurrence for patients undergoing NAC versus patients undergoing surgery first. METHODS A total of 1176 patients with clinically node negative breast cancer underwent SNB from 2007 to 2013. Clinicopathologic and survival data were reviewed and comparisons made between patients receiving NAC and those undergoing surgery first. Lymphatic mapping was performed with both radioactive colloid and blue dye. Patients with negative SN for metastasis were followed without axillary lymph node dissection (ALND). Patients with metastases to a SN underwent ALND. RESULTS Of the patients, 180 (15.3%) underwent SNB following NAC and 996 (84.7%) underwent surgery first. SN identification rates were 98.3% in the NAC group and 98.9% in the surgery first group. 152 (84.4%) patients in the NAC group and 877 patients (88.1%) had negative SNs. At median follow-up of 49.2 months, 1 patient (0.007%) in the NAC group and 3 patients (0.003%) in surgery first group had axillary lymph node recurrences. There were not any significant differences between the NAC group and the surgery first group. CONCLUSION The SNB following NAC in patients with node-negative breast cancer has a low axillary recurrence rate and could be acceptable. Citation Format: Hiroko Nogi, Ken Uchida, Makiko Kamio, Kumiko Kato, Yasuo Toriumi, Hiroshi Takeyama. Long-term follow-up of the node-negative breast cancer patients before treatment evaluated with sentinel node biopsy alone following neoadjuvant chemotherapy [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 P2-01-19.

Usefulness of contrast-enhanced ultrasonography for intracystic breast tumors: a report of three cases

Cystic breast lesions are caused by a wide spectrum of breast diseases and can range from simple cysts to malignant tumors. Ultrasonography is a good tool for evaluation of the morphology and vascularity of cystic breast lesions. We report three patients in whom contrast-enhanced ultrasonography (CEUS) was used to evaluate intracystic tumors. One of the three patients was diagnosed with intracystic papilloma. Compared with conventional ultrasonography alone, CEUS more clearly demonstrated that the solid component within the mass was lobulated with a narrow base. The other two patients were diagnosed with intracystic papillary carcinoma, and CEUS clearly revealed the presence of widely elevated solid components within both masses, suggesting malignancy. Therefore, CEUS simplified morphological evaluation by enhancing the solid components within the cystic masses.

Abstract P4-06-11: HER2 type DCIS acquires histological diversity by p53 mutation

[Introduction] It is widely known that breast cancer is a heterogeneous disease of various phenotypes and biological characteristics. Several studies have identified distinct subtypes of invasive ductal carcinoma by gene expression profiling or staining pattern of estrogen receptor (ER), progesterone receptor (PR), and HER2/neu protein. Ductal carcinoma in situ (DCIS) is a non-invasive form of breast cancer, and is thought to be the precursor of invasive breast cancer. One of the most well-known hypothesis shows that breast cancer occurs in a normal terminal duct lobular unit, and a histological continuity exists between the precursor and breast cancer. The aim of this analysis is to reveal how DCIS acquires histological diversity and progresses to invasive ductal carcinoma. We hypothesized that p53 gene mutation is responsible for this acquisition of histological diversity, and performed immunohistochemical analysis to pursue this hypothesis. [Material and methods] The participants of this study were taken from a database established by Jikei University School of Medicine. Between the period of April 2000 and April 2011, 84 cases of pure DCIS patients underwent operation. Patients were classified into 4 subtypes by a combination of hormone receptor (HR) status and HER2 status. The status of ER, PR, HER2, and p53 was determined by immunohistochemical staining. Tumors with Allred score above 2 ER / PR nuclear staining were classified as ER / PR positive. Tumors with HER2 membranous staining equivalent to 3+ intensity with Hercep test in more than 30% of the cells were scored as overexpression. p53 was defined as positive when nuclear staining was equal to or greater than 10%. We evaluated histological nuclear atypia to assess histological diversity of DCIS. When DCIS included different type of atypia cells more than 10%, we judged the histological diversity as positive. [Results] Patient characteristics are presented in table1. The classifications by the immunohistochemical subtype of DCIS are as follows: HR+/HER2- 62 cases (74%), HR+/HER2+ 5 cases (6%), HR-/HER2+ 13 cases (15%), HR-/HER2- 4 cases (5%). p53 expression in HR-/HER2+ and HR-/HER2- subtype was significantly higher than HR+/HER2- and HR+/HER2+ subtype (p The frequency of histological diversity by nuclear atypia was high in HR+/HER2- and HR-HER2+ subtype (p = 0.009). In HR-HER2+ subtype, p53 expression was associated with histological diversity (p = 0.021). But in HR-HER2+ subtype, there was no association with p53 expression and histological diversity. [Conclusion] This analysis demonstrates that HR-HER2+ subtype DCIS acquires histological diversity by p53 mutation. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-06-11.