Kento Kurata

Tumour-infiltrating CD8 to FOXP3 lymphocyte ratio in predicting treatment responses to neoadjuvant chemotherapy of aggressive breast cancer

Tumour‐infiltrating lymphocytes (TILs) can be used to monitor the immune response, and are important in predicting treatment responses and outcomes for various types of cancer. Recently, specific TIL subsets have been reported to be clinically useful in predicting treatment responses. The CD8+/FOXP3+ TIL ratio (CFR) may be a more sensitive indicator for monitoring immune function. This study investigated the clinical significance and value of CFR as a biomarker to predict treatment responses to neoadjuvant chemotherapy for breast cancer.

Clinical verification of sensitivity to preoperative chemotherapy in cases of androgen receptor-expressing positive breast cancer.

Background:Triple-negative breast cancer (TNBC) patients testing positive for androgen receptor (AR) expression are thought to be chemotherapy resistant, similar to other hormone receptor-positive breast cancers; however, this has not been substantially validated in the clinic. In this study, we investigated the association between chemotherapy sensitivity and AR expression in patients treated with neoadjuvant chemotherapy (NAC) using standardised chemotherapy criteria and regimens.Methods:A total of 177 patients with resectable early-stage breast cancer were treated with NAC. Oestrogen receptor, progesterone receptor, HER2, Ki67 and AR status were assessed immunohistochemically.Results:Sixty-one patients were diagnosed with TNBC; AR expression was identified in 23 (37.7%), which was significantly less common than that found in non-TNBC patients (103 of 116; 88.8%; P<0.001). The rate of pathological complete response after NAC was significantly lower (P=0.001), and disease recurrence was more common (P=0.008) in patients with AR-positive compared with those with AR-negative TNBC. In TNBC cases, as expected, the non-recurrence period in cases that were negative for AR expression was significantly extended (P=0.006, log-rank).Conclusions:Androgen receptor expressions may be useful as biomarkers to predict treatment responses to NAC in TNBC. Moreover, induction of a change in subtype to the AR-negative phenotype was observed after NAC.

Growth arrest by activated BRAF and MEK inhibition in human anaplastic thyroid cancer cells

Anaplastic thyroid cancer (ATC) is a rare malignancy that progresses extremely aggressively and often results in dismal prognosis. We investigated the efficacy of inhibiting the activated RAS/RAF/MEK pathway in ATC cells aiming to clarify the mechanism of effect and resistance. Four human ATC cell lines (ACT-1, OCUT-2, OCUT-4 and OCUT-6) were used. OCUT-4 had a BRAF mutation. OCUT-2 had both BRAF and PI3KCA mutations. ACT-1 and OCUT-6 had wild-type BRAF and NRAS mutations. The effects of dabrafenib, a selective inhibitor of the BRAFV600E kinase, and trametinib, a reversible inhibitor of MEK activity, were investigated. Dabrafenib strongly inhibited the viability in BRAF mutated cells by demonstrating G0/G1-arrest via the downregulation of MEK/ERK phosphorylation. Upregulated phosphorylation of MEK was observed in RAS mutated cells after dabrafenib treatment and caused VEGF upregulation, but was not related to the cellular proliferation. Trametinib inhibited the cellular viability to variable degrees in every cell by downregulating ERK phosphorylation. Dual blockade by both inhibitors demonstrated clear cytostatic effect in all the cells. OCUT-4 showed the weakest sensitivity to trametinib, no additional effect of either inhibitor in combination with the other, and an increase of SNAI1 mRNA expression after treatment with inhibitors, suggesting a mechanism for resistance. Our findings demonstrated the efficacy of a mutation-selective BRAF inhibitor and a MEK inhibitor in human ATC cells in a genetic alteration-specific manner.

Ambulatory sentinel lymph node biopsy preceding neoadjuvant therapy in patients with operable breast cancer: a preliminary study.

BackgroundSentinel lymph node biopsy (SNB)-oriented stepwise treatment under local anesthesia has been performed in the outpatient-ambulatory setting in patients receiving neoadjuvant therapy (NAT). We retrospectively reviewed our preliminary experience of ambulatory SNB in breast cancer patients scheduled to undergo NAT to evaluate the usefulness and feasibility of this method as a minimally invasive, stepwise treatment protocol.MethodsWe retrospectively identified 56 patients with breast cancer without obvious nodal involvement who were scheduled to receive NAT before breast surgery. SNB was performed under local anesthesia in an ambulatory outpatient setting before the initiation of NAT.ResultsThe average number of removed sentinel lymph nodes was 1.9. Identification of the sentinel node was possible in all cases, and macrometastasis was observed in six cases (10.7%). Micrometastasis was observed in five cases, while isolated tumor cells were noted in six cases. There were no delays in the initiation of NAT as a result of complications of SNB.ConclusionsThis pilot study demonstrated the safety and feasibility of ambulatory SNB prior to NAT. Further studies are warranted to assess the strict indications, patient satisfaction, and medical economics of this procedure.

Identification of tumour-reactive lymphatic endothelial cells capable of inducing progression of gastric cancer

Background:Tumour cells and stromal cells interact in the tumour microenvironment; moreover, stromal cells can acquire abnormalities that contribute to tumour progression. However, interactions between lymphatic endothelial cells (LECs) and tumour cells are largely unexamined. In this study, we aimed to determine whether tumour-specific LECs inhabit the tumour microenvironment and examine their influence on this microenvironment.Methods:We isolated normal LECs (NLECs) from a non-metastatic lymph node and tumour-associated LECs (TLECs) from cancerous lymph nodes. We examined proliferative and migratory potency, growth factor production, and gene expression of each type of LEC. Moreover, we developed a co-culture system to investigate the interactions between gastric cancer cells and LECs.Results:When compared with NLEC, TLECs had an abnormal shape, high proliferative and migratory abilities, and elevated expression of genes associated with inflammation, cell growth, and cell migration. NLECs co-cultured with gastric cancer cells from the OCUM12 cell line acquired TLEC-like phenotypes. Also, OCUM12 cells co-cultured with TLECs expressed high levels of genes responsible for metastasis.Conclusions:Our results demonstrated that LECs interacted with tumour cells and obtained abnormal phenotypes that could have important roles in tumour progression.

Abstract 5127: Clinical verification of antitumor autoimmune response in eribulin chemotherapy for breast cancer

Background: The immune environment of the cancer tissue not only influences the response to immunotherapy, but also the response to and outcomes after other anti-cancer therapies. Thus, the importance of regulating and improving the cancer immune microenvironment is increasingly being recognized. Tumor-infiltrating lymphocytes (TILs) can be used to monitor the immune response and are important in predicting treatment responses and outcomes in various types of carcinoma. On the other hand, it was reported that eribulin suppresses epithelial-mesenchymal transition (EMT) in basic studies, and we investigated it with clinical samples. In addition, it is thought that antitumor autoimmune response relates to suppression of EMT. In the present study, we investigated clinically antitumor autoimmune response in breast cancer with eribulin chemotherapy. Materials and Methods: Eribulin chemotherapy was administered to 52 patients with locally advanced or metastatic breast cancer. Samples of cancer tissue obtained before and after treatment were available from 10 patients. We evaluated response rate (RR) and analyzed protein expression by immunostaining of specimens before and after treatment. We evaluated antitumor autoimmune response by PD-1, CD8, FOXP3 expression in stromal TILs and PD-L1, PD-L2 expression in cancer cells. And, we examined the relation between the expression of these immune related molecules and EMT markers (evaluated by E-cadherin, N-cadherin, Vimentin expression). Results: In 11 patients (21.2%), it was possible to excise the lesion after treatment. In 10 evaluable cases without one pathological complete response (pCR) case, PD-1, PD-L2, and FOXP3 expression turned to negative in 5 cases, PD-L1 expression turned to negative in 6 cases, and CD8 expression turned to positive in 3 cases before and after treatment with eribulin chemotherapy. Looking at the relation between the transition in this protein expression and therapeutic effect, cases observed with negative conversion in PD-L1, FOXP3 expression had significantly high RR (p = 0.024) (p = 0.004). Among high RR cases, CD-8 expression was increased, the PD-1, PD-L1 and FOXP3 expression were remarkably reduced. Furthermore, we found a inverse correlation between PD-L1, FOXP3 expression turning to negative and E-cadherin expression turning to positive (p = 0.024) (p = 0.004). Conclusions: In the chemotherapy with eribulin, PD-L1 and FOXP3 expression turning to negative before and after treatment suggested improvement of the tumor immune microenvironment, and this mechanism related to suppression of EMT. Citation Format: Wataru Goto, Shinichiro Kashiwagi, Yuka Asano, Kento Kurata, Tamami Morisaki, Satoru Noda, Tsutomu Takashima, Naoyoshi Onoda, Sayaka Tanaka, Masahiko Ohsawa, Masaichi Ohira, Kosei Hirakawa. Clinical verification of antitumor autoimmune response in eribulin chemotherapy for breast cancer. [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 5127.

Nonfunctional adrenocortical carcinoma initially presenting as retroperitoneal hemorrhage.

BackgroundAcute adrenal hemorrhage is an uncommon entity. Although trauma is the most common cause of adrenal hemorrhage, non-traumatic etiologies have also been reported. We report an unusual case of a spontaneously ruptured adrenocortical carcinoma that initially presented as a critical massive retroperitoneal hemorrhage. The case was treated successfully using a combination of emergency interventional radiology and elective surgery.Case presentationA 47-year-old woman was transported to our hospital because of the sudden onset of severe pain in her left lower back. The shadow of a tumor-like soft mass accompanied by bleeding was observed in the upper pole of the left kidney, together with vascular leakage from the middle suprarenal artery on computed tomography. Transcatheter embolization of the left middle adrenal artery was administered based on a diagnosis of acute adrenal hemorrhage. Further observation indicated that the bleeding was caused by rupture of an adrenocortical carcinoma. Left adrenalectomy was subsequently carried out via laparotomy.ConclusionsWe experienced an unusual case of acute massive adrenal hemorrhage caused by the rupture of a non-functional adrenocortical carcinoma, which was treated successfully by ambulatory transcatheter embolization therapy and elective surgery.

173PTLE3 IS A USEFUL MARKER FOR PREDICTING THE THERAPEUTIC EFFECT OF ERIBULIN CHEMOTHERAPY FOR TRIPLE-NEGATIVE BREAST CANCER

ABSTRACT Aim: The recently developed reagent, eribulin mesylate (eribulin), is a microtubule dynamics inhibitor with a mechanism of action that differs from that of taxanes and vinca alkaloids. This drug is considered to be a promising chemotherapeutic agent for the treatment of locally advanced or metastatic breast cancer (MBC). In this study, we investigated predictive factors with respect to the therapeutic effect of eribulin chemotherapy among variables such as b-tubulin class III, GSTP1 and TLE3, which have previously been reported to be predictive factors of the therapeutic effect of taxanes, in an aim to identify possible biomarkers for predicting the efficacy of eribulin. Methods: The subjects included 52 patients with MBC who underwent chemotherapy using eribulin. The median follow-up time was 431 days (range, 50-650 days). The overall response rate (ORR), clinical benefit rate (CBR), disease control rate (DCR), overall survival (OS), time to treatment failure (TTF) and progression-free survival (PFS) were calculated regarding the efficacy of this regimen. Moreover, breast cancer was classified into intrinsic subtypes according to the status of the ER, PR, HER2 and Ki67 expression. Results: The clinical effects were as follows: overall ORR= 34.6%, CBR = 44.2%, DCR= 51.9%; median OS = 334 days; median TTF = 81 days; and median PFS = 275 days. TLE3, b-tubulin class III and GSTP1 were expressed in 24 cases, 21 cases and 24 cases, respectively, among the 52 patients investigated. The expression of TLE3 was found significantly more frequently in the TNBC lesions than in the non-TNBC lesions (p = 0.030). The patients with TLE3-negative tumors experienced significantly poorer outcomes in terms of progression-free survival when comparing the prognosis within the group of patients with triple-negative breast cancer (TNBC) lesions (p = 0.011). Based on a multivariate logistic regression analysis including 22 patients with TNBC also showed that a positive TLE3 expression significantly correlated with a better progression-free survival (p = 0.037). Conclusions: Our findings suggest that TLE3 is a useful marker for predicting the therapeutic effect of eribulin chemotherapy for TNBC. Disclosure: All authors have declared no conflicts of interest.

Predictive factors for the occurrence of four or more axillary lymph node metastases in ER-positive and HER2-negative breast cancer patients with positive sentinel node: A retrospective cohort study

INTRODUCTION Patients with four or more axillary lymph node metastases have benefited from postmastectomy radiotherapy to the supraclavicular region. However, when metastatic sentinel nodes (SNs) are present, information regarding the total number of node metastases cannot be obtained if axillary lymph node dissection (ALND) is omitted from the treatment protocol. It is important to determine the indication for additional chemotherapy in ER-positive and HER2-negative breast cancer patients. We investigated the predictive factors for the occurrence of four or more metastases in patients with ER-positive and HER2-negative breast cancer in the presence of macrometastasis in the SNs. METHODS We reviewed 83 patients with ER-positive and HER2-negative breast cancer, who had macrometastasis in the SN and had undergone ALND. The clinicopathological findings and prognosis between patients with pN1 disease and those with pN2 disease were also compared. RESULTS Nineteen percent of patients had pN2-3 disease. The predictive factor for poor prognosis in these patients was the presence of pN2-3 disease. The independent predictive factors for pN2-3 disease were the T stage and the ratio of the number positive SNs to the number of removed SNs (SN ratio). Patients with both T2 tumors and a high SN ratio had a 50% risk of having pN2-3 disease. CONCLUSION The presence of four or more metastases was found to be the strongest prognostic factor in ER-positive and HER2-negative breast cancer patients with macrometastasis in the SN. The T stage and SN ratio determined before surgery or during surgery were useful in predicting pN2-3 disease in these patients.

Abstract 3939: Clinical significance of expression of androgen-receptor splice variant-7 (AR-V7) in neoadjuvant chemotherapy for breast cancer

Background: Triple-negative (TN) breast cancer (BC) patients testing positive for androgen receptor (AR) expression are thought to be chemotherapy resistant, similar to other hormone receptor-positive (HR) BCs. On the other hand, prostate cancer is a hormone-dependent malignant tumor like breast cancer, it has been reported that chemotherapy is more likely to be effective if there is an abnormality in androgen-receptor splice variant-7 (AR-V7). In an immunohistochemical study, as well, AR-V7 expression was reportedly a poor prognostic factor in castration-resistant prostate cancer (CRPC). In this study, we investigated the association between chemotherapy sensitivity and AR-V7 expression in patients treated with neoadjuvant chemotherapy (NAC) using standardized chemotherapy criteria and regimens. Materials and Methods: A total of 177 patients with resectable early-stage breast cancer were treated with NAC. ER, PgR, HER2, Ki67 and AR-V7 status were assessed immunohistochemically. Results: In the 43 AR-V7 expression positive group, compared to the 133 negative group, TNBC (p Conclusion: These findings show that AR-V7 expression is a therapeutic effect predictive marker in BCNAC, and indicates particularly high chemotherapy sensitivity in TNBC. Citation Format: Yuka Asano, Shinichiro Kashiwagi, Wataru Goto, Kento Kurata, Tamami Morisaki, Satoru Noda, Tsutomu Takashima, Naoyoshi Onoda, Sayaka Tanaka, Masahiko Ohsawa, Masaichi Ohira, Kosei Hirakawa. Clinical significance of expression of androgen-receptor splice variant-7 (AR-V7) in neoadjuvant chemotherapy for breast cancer. [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 3939.