Kaisuke Miyamoto

Abstract 4138: Immunological profile of metastatic or recurrent breast cancer patients

Background: We previously reported about immune suppression in breast cancer patients at this meeting. In brief, tumor tissue specimens and peripheral blood mononuclear cells (PBMC) were analyzed in 50 early or advanced breast cancer (BC) patients. To compare between 38 early and 12 advanced BC cases, CD163-positive tumor cells and CCR4-positive tumor cells were detected more frequently in advanced cases than in early cases. Regulatory T (Treg) cells in PBMC significantly increased in percentage of the population in 37 BC patients than in 21 healthy volunteers (AACR 2011). In addition, several cytokines were examined in that cohort and plasma IL-17A had significantly higher levels in early BC than in advanced BC (AACR 2013). Then, we examined their prognosis and immunological profile. Patients and methods: Treg cells were examined by counting CD4+CD25highCD127low/-cells in PBMC with flow cytometry analysis. Immunohistochemical evaluation of tumor specimens was performed with monoclonal antibodies of HLA-ABC and DR, CD56, CD68, CD83, CD163 and CCR4. The number of stained cells was analyzed using a semiquantitative ordinal scale ranging from 0 to 3 (0, +/-, ++, +++). Human IL-2, IL-4, IL-6, IL-10, TNF, INFγ and IL-17A were measured using cytometric beads array system. Most patients received chemotherapy, hormonal therapy, and/or anti-HER2 therapy on the basis of intrinsic subtype and breast irradiation after breast-conserving surgery. Results and discussions: At the median follow-up of 7 years after blood sample collection, only 2 operable patients relapsed and 3 patients including 2 cases of stage IV died of disease. Of 5 cases of stage IV or recurrent BC, CD163 and/or CCR4 were strongly stained positive in tumor cells. There were significant differences of staining intensity in CD163 and CCR4-positive tumor cells between those cases and the rest 45 cases (p Conclusion: Targeted therapy against M2 macrophage or CCR4 is considered as a promising strategy of advanced breast cancer. Citation Format: Shigeru Imoto, Takayuki Ueno, Hirotsugu Isaka, Hiroki Ito, Kaisuke Miyamoto, Tomohiro Chiba, Hiroshi Kamma. Immunological profile of metastatic or recurrent breast cancer patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4138.

Abstract 3484: Analysis of in situ expression of hormone receptors and proliferation marker at a single cell level in breast cancer tissues

Background: Hormone receptors and proliferation markers are critical parameters for treatment selection of breast cancer patients. The expression of different parameters are currently assessed separately in different tissue sections but it is unclear how different parameters are co-expressed in a single cell in breast cancer tissues. Samples and Methods: Breast cancer tissues from fifty-one patients with ER-positive HER2-negative breast cancer were analyzed. Expressions of ER and PgR were assessed in association with Ki67 using dual fluorescence immunohistochemistry with specific antibodies: SP-1 (abcam, Tokyo), 1E2 (Roche Diagnostics GmBH, Germany), and MIB1 (Dako Japan, Tokyo), respectively. More than 500 cancer cells were assessed in each tissue. Expression levels of each marker in a single cell were semi-quantitatively assessed by MetaMorph image analyzer (Molecular Devices Japan, Tokyo). All statistical analyses were performed using JMP ver.8.01 (SAS Institute Japan, Tokyo). Results: To validate the system, Ki67 LI in breast cancer tissues were compared between this system and the regular DAB system. The two systems showed a good concordance (p Conclusion: PgR status in Ki67-positive proliferating cells is associated with histological grade in ER positive HER2-negative breast cancers. Our results suggest that different driving systems give different expression patterns of PgR and Ki67 at a single cell level, which may distinguish between luminal A and luminal B cancers. Citation Format: Takayuki Ueno, Hirotsugu Isaka, Hiroki Itoh, Kaisuke Miyamoto, Manami Kitamura, Shigeru Imoto. Analysis of in situ expression of hormone receptors and proliferation marker at a single cell level in breast cancer tissues. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3484.

Abstract 1083: Host-tumor immune response for early breast cancer patients treated with radiofrequency ablation therapy

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA [Background] We demonstrated immune suppression of regulatory T (Treg) cells in preoperative breast cancer (BC) patients at AACR 2011. Last year we also reported at this meeting that Th17 cells might have some potential to control tumor progression in the same cohort. Modern treatment of operable BC consists of breast surgery, drug therapy and radiation therapy. [Aim] To explore new strategy of BC treatment, we conducted a phase II study on radiofrequency ablation (RFA) in early BC between 2009 and 2013. Twenty patients were enrolled. To examine host-tumor immune response under thermal ablation therapy, Treg cells in peripheral blood mononuclear cells (PBMC) and plasma IL-17A were measured. [Patients and methods] Blood samples were collected from 13 patients before RFA and at the time of 1 month after RFA. Treg cells were examined by counting CD4+CD25highCD127low/-cells in PBMC. Plasma IL-17A was measured using cytometric beads array system. Statistical significance was analyzed using Mann-Whitney U-test. [Results] As of November 2013, all patients were free of recurrence. The percentages of Treg cells had no significant change in 13 patients before and after RFA (2.8% versus 2.7% of CD4-positive cells at mean value). In addition, there was no statistical difference in levels of plasma IL-17A before and after RFA (76 pg/ml versus 116 pg/ml at mean level). [Discussion] Unexpectedly, RFA had no favorable immune response in BC patients, because we observed significant decrease in percentages of Treg cells after breast surgery. The reason is that most patients had T1 tumor and Luminal A-like BC. They had lower percentages of Treg cells in comparison to those of tumor-bearing patients reported previously (2.8% versus 4.2%). [Conclusion] RFA is a promising alternative to breast surgery. However, it may make little effect on host-tumor immune response in favorable subtype of early BC. Citation Format: Shigeru Imoto, Noriko Nakatsugawa, Takayuki Ueno, Hiroki Ito, Kentaro Imi, Kaisuke Miyamoto, Manami Kitamura, Hirotsugu Isaka, Tetsuya Nakatsura. Host-tumor immune response for early breast cancer patients treated with radiofrequency ablation therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1083. doi:10.1158/1538-7445.AM2014-1083

Abstract 450: Host-tumor immune response for breast cancer patients.

We demonstrated immune suppression of regulatory T cells (Treg) and M2 macrophage in breast cancer (BC) patients at AACR 2011. In summary, to compare between 38 early and 12 advanced BC cases, CD163-positive tumor cells and CCR4-positive tumor cells were detected more frequently in advanced cases than in early cases. Treg cells in PBMC significantly increased in percentage of the population in 37 BC patients than in 21 healthy volunteers. To explore host-tumor immune response, several cytokines were examined in the same cases. Plasma from 32 early and 9 advanced BC cases was preoperatively obtained. Human IL-2, IL-4, IL-6, IL-10, TNF, INFγ, and IL-17A were measured using cytometric beads array system. Statistical significance was analyzed using Mann-Whitney U-test. Plasma IL-17A had significantly higher levels in early BC than in advanced BC (103 pg/ml v.s. 51 pg/ml at mean level). Th17 cells may have some potential to control tumor progression in BC. In conclusions, induction of host-tumor immune response due to chemotherapy and anti-HER2 therapy and release from immune suppression such as CCR4 monoclonal antibody are important issues to explore a new strategy for BC patients. Citation Format: Shigeru Imoto, Noriko Nakatsugawa, Hirotsugu Isaka, Hiroki Ito, Kentaro Imi, Kaisuke Miyamoto, Tetsuya Nakatsura. Host-tumor immune response for breast cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 450. doi:10.1158/1538-7445.AM2013-450

Paradigm shift in axilla surgery for breast cancer patients treated with sentinel node biopsy

BackgroundSentinel node biopsy (SNB) is a standard technique for the diagnosis of regional lymph node metastases in clinically node-negative breast cancer patients. In the case of pathologically negative sentinel lymph nodes (SLN), axillary lymph node dissection (ALND) can be avoided.MethodsRecent clinical studies on SNB in breast cancer were reviewed regarding the pathological and molecular diagnosis of SLN, the tools used to predict non-SLN metastases, the prognostic significance of isolated tumor cells (ITC) and micrometastases (MIC), and axilla surgery.ResultsITC or MIC in SLN was associated with worse survival in patients treated with SNB alone or SNB followed by ALND. However, this effect was limited and adjuvant therapy improved survival. If T1 and one SLN-positive breast cancer patients are treated with whole-breast irradiation and adjuvant therapy, additional ALND may not be necessary.ConclusionsSNB without ALND can be adopted for patients with a small number of SLN metastases. Although the lack of apparent regional lymph node recurrence, similar to tumor dormancy, cannot be fully explained, ALND should be performed in cases that are highly suspected to be non-SLN metastases.

P1-01-11: CD4+CD25highCD127low/− Regulatory T Cells Have Immunosuppressive Function in Patients with Breast Cancer.

CD4 + CD25 high Foxp3 + regulatory T cells play a central role in self-tolerance and suppress antitumor immune response. Recent studies have shown that low levels of the IL-7 receptor (CD127) are expressed on CD4 + CD25 high Foxp3 + regulatory T cells surfaces and the expression of CD127 is inversely correlated with the suppressive function of CD4 + CD25 high regulatory T cells. We evaluated the frequency of CD4+CD25 high CD127 low/− T cells in peripheral blood lymphocytes of 72 breast cancer patients and 21 healthy volunteers. The expression of Foxp3 mRNA was inversely correlated with the CD127 expression and correlated with CD25 expression. The frequency of CD4+CD25 high CD127 low/− T cells in breast cancer patients was significantly higher than that of healthy volunteers (p=0.0045). CD4+CD25 high CD127 low/− T cells were increased in hormone receptor negative patients and HER2 positive patients, but no statistical significant was observed in stage progression. In addition, the frequency of CD4+CD25 high CD127 low/− T cells decreased after curative resection in breast cancer patients (p=0.005). CD4+CD25 high CD127 low/− T cells also suppress proliferation of autologous CD4 + CD25 − helper T cells and CD8 + cytotoxic T cells in CFSE-based cell proliferation assay. These findings suggest that CD4+CD25 high CD127 low/− T cells a useful marker for regulatory T cells in breast cancer. Circulative peripheral regulatory T cells may participate in immune tolerance to breast carcinoma. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-01-11.