Search

Background: Expression of immune checkpoint receptors (ICR) on tumor infiltrating lymphocytes (TILSs) is associated with better response to immunotherapies via immune checkpoint inhibitors. Therefore, we investigated various ICR expressions on TILs in patients with locally advanced triple negative breast cancer (TNBC) after neoadjuvant chemotherapy (NAC). Methods: Expressions of ICRs were examined immunohistochemically by staining surgical specimen (n=61) using specific monoclonal antibodies for PDL-1, PD-1, TIM-3, LAG-3, CTLA-4. Positivity was defined staining >1% on TILs. Results: Median age was 49 (24-76) years. The majority patients were clinically T3-4 (n=31, 50.8%), and clinically N1-3 (n=58, 95.1%) before NAC. Of those, 82% were found to have CTLA-4 positivity, whereas TILs associated positivites for PD1, PDL-1, LAG3 and TIM-3 were 62.3%, 50.9%, 26.2%, 68.9%. High expression of CTLA-4 was found to be associated with a better chemotherapy response (OR=7.94, 95%CI: 0.9-70.12, p=0.06), whereas TIM-3 positivity was contrarly associated with a worse chemotherapy response (OR=0.253, 95%CI: 0.066-0.974, p=0.047) as measured by MDACC Residual Cancer Burden Index. At a 47-month follow-up, patients with ypN0 disease (DFS; HR=0.31, 95% CI: 0.12-0.83, p=0.02 and DSS; HR=0.21, 95% CI:0.07-0.62, p=0.005) and CTLA-4 high expression on TILs (DFS; HR=0.38, 95% CI=0.17-0.85, p=0.019 and DSS; HR=0.34, 95% CI:0.15-0.78, p=0.01) were found to have improved survival. Conclusions: These findings demonstrate that CTLA-4, PD-1, PDL-1 and TIM-3 were highly expressed in TNBC after NAC. Our results more favor an immuncheckpoint inhibitor therapy via CTLA-4 alone or in combination with other immune check point inhibitors against PDL-1 and/or TIM-3 in addition to NAC in advanced TNBC. Citation Format: Neslihan Cabioglu, Semen Onder, Gizem Oner, Hüseyin Karatay, Mustafa Tukenmez, Mahmut Muslumanoglu, Abdullah Igci, Yesim Eralp, Adnan Aydiner, Pinar Saip, Ekrem Yavuz, Vahit Ozmen. Tim3 expression on tumor infiltrating lymphocytes is associated with poor response to neoadjuvan chemotherapy in patients with locally advanced triple negative breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS4-32.

BACKGROUND: Omitting axillary lymph node dissection (ALND) following sentinel lymph node biopsy (SLNB) in patients with locally advanced breast cancer (LABC) after neoadjuvant chemotherapy (NAC) is still controversial. In this study, we evaluated factors affecting local recurrence and outcome in patients with LABC, who underwent SLNB alone after NAC. METHODS: Between 2004 to 2018, 320 patients with clinically node-positive LABC who received NAC and underwent SLNB alone after negative axillary staging were analyzed. All patients had breast and/or regional nodal irradiation. RESULTS: Median age was 46 (23-70). Of those, 228 patients had ypN0 disease (71.25%), whereas 92 patients had ypN(+) disease including 19 (20.6%) isolated tumor cells (ITC), 33 micrometastasis (35.9%) and 40 macrometastases (43.5%). At a median follow-up of 37 months (24-172), one patient (0.3%) with macrometastatic SLN having extracapsular extension was found to have locoregional recurrence at the 60th month. Five-year disease-free survival (DFS) and disease specific survival (DSS) rates were found as 87% and 95%, respectively. Patients with cT3&4 (HR=2.22, 95% CI; 1.07-4.62), non-luminal molecular pathology (HR=2.71, 95% CI, 1.23-5.97), and non-pCR in the breast (HR=2.21, 95% CI, 0.94-5.17) were found to have an increased HR compared to others in 5-year DFS. However, no significant differences could be found between patients ypN0, ypN-ITC&micrometastasis and ypN-macrometastasis regarding 5-year DFS and DSS rates. CONCLUSIONS: ALND could be safely avoided in selected patients with LABC who underwent SLNB after NAC having cT1-2, luminal pathology, breast and/or nodal pCR or low volume nodal disease, as long as axillary radiotherapy is provided. Citation Format: Neslihan Cabioglu, Hasan Karanlık, Mahmut Muslumanoglu, Guldeniz Karadeniz Çakmak, Didem Can Trablus, Mustafa Tukenmez, Yeliz E. Ersoy, Cihan Uras, Baha Zengel, Selman Emiroğlu, Ayfer Kamalı Polat, Levent Yeniay, Enver Ozkurt, Halil Kara, Kamuran Ibis, Adnan Aydiner, Vahit Ozmen, Abdullah Igci. Sentinel lymph node biopsy alone in locally advanced breast cancer after neoadjuvant chemotherapy: Turkish multicentric neosenti-turk MF-18-02-study [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS1-26.

Background: Whether axillary lymph node dissection (ALND) following sentinel lymph node biopsy (SLNB) could be spared in patients with initially clinically positive axilla after neoadjuvant chemotherapy (NAC) is still controversial even though recent studies indicate that axillary recurrence seems to be a rare event. Our aim is to find out whether omitting ALND could be oncologically safe in patients undergoing SLNB after NAC. Material and Methods: Of patients presented with c T1-4N1-3M0 disease, those undergoing SLNB after NAC were included in the prospective multicentre registry trial " MF18-03/BHWG" (ClinicalTrials.gov/NCT04250129). Cases with inflammatory breast cancer, distant metastases, pregnancy, bilateral breast cancer, or other cancers and those without adjuvant nodal radiotherapy were excluded from the study. The end points of the present report are the axillary nodal recurrence (AR) and locoregional recurrence (LRR) rates at a median follow-up more than 2 years, and determine factors associated with AR and LRR . The locoregional recurrences included ipsilateral, and contralateral axillary recurrences, infra-and supraclavicular recurrences, and recurrences in the mammaria interna region. Results: Between January 2018 to January 2021, 2358 patients with cN(+) disease, who became cN0 after NAC, and underwent SLNB, were analyzed. Median age was 47 (range, 21-86). Of those, the majority of patients had cT1-2 (80.5%) and N1 (80.3%) disease. Following NAC, half of the patients (50%) had breast conserving surgery, whereas the remaining half had mastectomy (50%). Of 2358 patients, 908 (38.5%) had ALND following SLN (ypN+, 85%) and 1450 (61.5%) underwent SLNB alone (ypN0, 72%). SLNB was performed by using the blue dye technique-alone in 66.6% of patients and by targeted axillary dissection in 659 patients (27.9%). Of those, 819 (34.8%) were HER2(+) and 373 (15.8%) were triple negative. The pCR rates for the axilla, breast and both for the axilla and breast were 50%, 35% and 28%, respectively. At a median follow-up time of 28 months (range, 12-62), the LRR, AR and isolated AR rates were 0.6% (n=14), 0.25% (n=6) and 0.13% (n=3), respectively. Furthermore, no significant difference could be found in LRR- and AR- rates between SLNB-alone and ALND groups regardless of the definitive nodal pathology (Table 1). Nodal recurrences were seen at a median of 12 months after the surgery. Of 6 cases with AR, 3 had synchronous local recurrences in breast, and 2 of them also had lung metastases in addition to local recurrence. All patients with AR were interestingly found to have HER2(+) or triple negative breast cancer at the initial diagnosis, and had residual invasive cancer in the breast surgical specimen. Logistic regression analyses revealed that patients with AR were significantly more likely to be younger than 45 (RR=7.81 ; 95% CI, 0.91-66.91) and have a cN2-3 (RR=4.1; 95% CI, 0.83-20.38), and non-luminal breast cancer (RR=12.47; 95% CI, 1.45-106.9) at the initial diagnosis (Table 2). Similarly, patients with LRR were more likely to present with cN2-3 disease (RR=3.09; 95% CI, 1.07-8.94) and non-luminal pathology (RR=6.27; 95%CI, 1.96-20.06) . Conclusion: This large prospective registry data also suggest that nodal recurrences can be detected at very low rates within 3 years after surgery in patients with clinically node-positive disease following NAC regardless of the extent of axillary surgery or nodal pathology as long as regional nodal radiation is provided. Since patients with early nodal recurrences have an agressive tumor biology with a potential of systemic recurrences, effective adjuvant systemic therapies should be considered in those with HER2(+) or triple negative residual breast cancer after surgery following adjuvant nodal radiation. Table 1. Local locoregoinal and systemic recurrences in cT1-4N1-3 patients with ypN0/ypN(+) diseases (n =2358) Table 2. Factors associated with axillary and locoregoinal recurrences (AR = axillary recurrences, LRR = locoregoinal recurrences, pCR = pathologic complete response) Citation Format: Neslihan Cabıoğlu, Hasan Karanlik, Mehmet Ali Gulcelik, Abdullah İgci, Mahmut Muslumanoglu, Havva Belma Kocer, Cihan Uras, Gokhan Giray Akgul, Mustafa Tukenmez, Serkan Ilgun, Didem Can Trabulus, Guldeniz Karadeniz Cakmak, Ahmet Dağ, Nilufer Yıldirim, Baha Zengel, Ebru Sen Oran, Kazim Senol, Halil Kara, Selman Emiroglu, M. Umit Ugurlu, Bulent Citgez, Yeliz Emine Ersoy, Atilla Celik, Ece Dilege, Yasemin Bolukbaşı, Niyazi Karaman, Gul Basaran, Aykut Soyder, Ayfer Kamali Polat, Gurhan Sakman, Serdar Ozbas, Ayse Altınok, Leyla Zer, Alper Akcan, Ibrahim Ali Ozemir, Levent Yeniay, N. Zafer Utkan, Lutfi Dogan, Mutlu Dogan, Mehmet Velidedeoglu, Beyza Ozcinar, Fazilet Erozgen, Abut Kebudi, Kemal Atahan, Vafa Valiyeva, Serdar Yormaz, Ali Sevinc, Cumhur Arici, Atilla Soran, Vahit Ozmen. PD15-01 AXILLARY NODAL RECURRENCE IS RARE IN PATIENTS WITH NODE-POSITIVE BREAST CANCER UNDERGOING SLNB FOLLOWING NEOADJUVANT CHEMOTHERAPY : EARLY RESULTS OF THE NEOSENTITURK-TRIAL/MF-18-03 [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 PD15-01.

Background: Residual nodal disease at the completion axillary lymph node dissection (ALND) has been reported in over 60% in patients with a positive sentinel lymph node biopsy (SLNB) after neoadjuvant chemotherapy (NAC). This study aimed to explore whether any clinical or histopathological characteristics of patients may be associated with a lower likelihood of having non-SLN metastasis at ALND. Methods: Between January 2004 to January 2021, 459 patients with cT1-4/N1-3 underwent ALND due to a positive SLNB after NAC. Demographic, clinical and histopathological characteristics of patients were analyzed to predict the non-SLN metastasis at ALND. Sentinel lymph node ratio was defined as the ratio of the number of positive SLN to the total excised SLNs. Results: Median age was 47 (21-84). Of those, the majority of the patients had cT1-2 (67.8%) and cN1 disease (78.1%), whereas 76.5% of tumors were invasive ductal carcinoma. The frequency of non-SLN metastases according to the tumor subtypes were 59.5% in luminal A, 73.8% in HER2(-) luminal B, 60.3% in HER2(+) luminal B, 66.7% in non-luminal HER2(+), and 66.7% in triple negative breast carcinoma. Of those with removal more than 2 SLNs, having 1 positive SLN, or breast pathologic complete response, or cT1-2 disease , or a SLN ratio FactorsOR(95%CI)p-valuecT0.009*1&20.45(0.25-0.82)3&4Reference (1)SLN Status0.006*ITC and micrometastasis0.24(0.09-0.65)MacrometastasisReference (1)SLN-Ratio (%) Citation Format: Neslihan Cabioglu, Hasan Karanlık, Abdullah Igci, Mahmut Muslumanoglu, Mustafa Tukenmez, Selman Emiroglu, Enver Ozkurt, Semen Onder, Pinar Saip, Yesim Eralp, Adnan Aydiner, Ekrem Yavuz, Vahit Ozmen. Factors predicting a lower likelihood of residual nodal disease in clinically-node positive patients undergoing sentinel node surgery after neoadjuvant chemotherapy for breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-01-07.

Background: Novel systemic therapies are under investigation for patients with triple negative breast cancer who are resistant to neoadjuvant chemotherapy or having a partial response to neoadjuvant chemotherapy. Immune check point inhibition or blocking other immunologic markers such as chemokine receptors might be alternative pathways for these chemotherapy-resistant patients. Therefore, we investigated various immune check point receptor expressions along with different chemokine receptors. Method: Expressions of immunological markers were examined immunohistochemically by staining archival tissue of mastectomy specimen (n=56) using specific monoclonal antibodies for PDL-1 (Ventana SP263 clone kit), CXCR4, CXCR5, CCR5, CCR7, CD73, and CD155. PDL-1 positivity was defined as membranous staining >1% in either tumor and /or stromal lymphocytes, whereas positivity of chemokine receptors and CD155 and CD73 were considered as cytoplasmic staining >50% and >5%, and >1%, respectively. Results: Median age was 47 (24-76) years. Of those, 31 were clinically T3-4 (55%), whereas almost all of them were N1-3 (96.4%) before neoadjuvant chemotherapy. All patients received anthracyclines&paclitaxel containing regimens, whereas 3 patients received additional carboplatin before definitive surgery. Of those, 30 patients (58%) were detected to be positive for PDL1 in tumor site and intratumoral lmyphocytes, whereas CXCR5 (41.1%, 23/56) , CCR5 (48.2%, 27/56), CCR7 (41.1%, 23/56), CD155 (30/56, 58%) and CD73 (44.6%, 25/56) were found to be highly expressed in tumors. However, 12.5% of patients have shown high expression of CXCR4. Patients with CXCR5, CCR5, CCR7, CD73, CD155 positivity were more likely found to give a worse chemotherapy response as measured by “Residual Cancer Burden Index”. Table 1.Associations between MD Anderson Residual Cancer Burden Index and Immunological Biomarker ExpressionsTumor characteristicsClass IClass 2&3P-valueCXCR4-negative8.3%91.7%0.999CXCR4-positive0%100% CXCR5-negative12.5%87.5%0.131CXCR5-positive0%100% CCR5-negative14.3%85.7%0.111CCR5-positive0%100% CCR7-negative12.5%87.5%0.131CCR7-positive0%100% CD73-negative13.3%86.70.117CD73-positive0%100% CD155-negative16%84%0.037CD155-positive0%100% PDL1-negative4.895.20.999PDL1-positive3.396.7% Median follow-up time was 36 months (7-177). Five-year disease-free survival (DFS) and disease specific survival (DSS) rates were found to be decreased in patients with CXCR4 (DFS: negative, 55% vs positive, 23%; p= 0.079 and DSS: negative, 52% vs positive, 23%; p=0.034) and CCR5 positivity(DFS: negative, 63.5% vs positive, 26%; p= 0.037 and DSS: negative, 61% vs positive, 25%; p=0.047), while no significant difference could be found in DFS and DSS rates in regards to PDL1, CXCR5, CCR7, CD73 and CD155 positivity. Conclusion: These results demonstrate that PDL-1, CD73, CD155 and chemokine receptors are highly expressed in patients with partial response to neoadjuvant chemotherapy. Furthermore, expressions of CXCR4 and CCR5 were found to be associated with poor prognosis in this cohort with triple negative breast cancer resistant to chemotherapy that would justify an addititional chemokine receptor inhibitor therapy. Citation Format: Cabioglu N, Onder S, Oner G, Karatay H, Tukenmez M, Muslumanoglu M, Igci A, Dinççag A, Aydiner A, Saip P, Ozmen V, Yavuz E. High expressions of CXCR4 and CCR5 are associated with poor prognosis in triple negative breast cancer resistant to neoadjuvan chemotherapy [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-06-26.

Background: Recent advances in tumor biology and immune system have identified check points modulating the anti-tumoral immune response. This study aimed to determine immune check point expressions patients with locally advanced breast cancer to explore any associations between clinical and prognostic factors to show effectiveness of targeted therapies for these molecules. Materials and Methods: This prospective study was carried out between September 2018 and November 2019 in the Department of General Surgery of Istanbul University Istanbul Faculty of Medicine Breast Unit. Twenty-four patients with LABC with a partial response to neoadjuvant chemotherapy (NAC) were included in the study. Tumor infiltrating lymphocytes (TILs) were isolated from the residual tumor burden separated from the surgical material with cell dissociation kit by using MACS tumor separation device. Expressions of immune checkpoints receptors including PD-1, LAG-3, TIM-3, TIGIT and CTLA-4 on CD8 T lymphocytes and Natural Killer (NK) cell subsets obtained from TIL were analyzed by using flow cytometry. Results: TILs of patients Conclusion: Our results suggest that patients with a younger age and advanced tumor burden are more likely to benefit from immunotherapeutical approaches using immune check point inhibitors. Furthermore, combination of systemic chemotherapy with immunotherapy may increase the pathologic response rate in breast cancer patients. Citation Format: Ayxan Abbasov, Esin Çetin, Neslihan Cabioglu, Baran Mollavelioglu, Semen Onder, Selman Emiroglu, Mustafa Tukenmez, Mahmut Muslumanoglu, Abdullah Igci, Gunnur Deniz, Vahit Ozmen. Differential expressions of immune check point receptors on tumor infiltrating lymphocytes after neo-adjuvant chemotherapy (NAC) in patients with locally advanced breast cancer (LABC) [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 947.

Introduction: High expression of immune checkpoint receptors in tumor microenvironment reduces antitumor immunity and cause immune evasion of tumor cells. In recent years, immunotherapy trials using PD-1 or PDL-1 inhibitors in advanced triple negative (TN) breast cancer evolved very rapidly. The differential expression of novel immune checkpoint receptors such as TIM-3, LAG-3 and TIGIT in addition to PD-1, and CTLA-4 on tumor infiltrating lymphocytes (TIL) in patients with early breast cancer was investigated. Material and methods: TIL were isolated by using a Tumor Dissociation Kit from fresh tumoral tissue. Flow-cytometric analyses were performed by using CD8, CD16, CD56, PD-1, CTLA-4, TIM-3, LAG-3 and TIGIT specific monoclonal antibodies on isolated TIL. Correlations were estimated between biological and clinical characteristics of tumors and demographic features of patients and flow cytometric findings. Results: Median age was 47 (range 28-68). There were 7 patients (35%) with HER2+ or triple negative tumors, whereas 13 patients (65%) had HER2 (-) luminal cancers. Our findings showed that patients younger than 45 years were more likely to express high levels of CTLA-4 (p=0.013) and TIGIT (p=0.007) on CD56+ natural killer (NK) cells and TIM-3 (p=0.043) on CD16+ lymphocytes (Table 1), whereas the other high expressions including LAG-3 (p=0,08) and TIM-3 (p=0.06) on CD56+ NK cells did not reach the statistical significance. Furthermore, patients with high Ki-67 proliferation index >35% were found to express higher CTLA-4 (p=0.011) on CD16+ lymphocytes. Patients with Stage II disease expressed higher levels of PD-1 (p=0.018) and LAG-3 (p=0.04) on CD8+ cytotoxic T lymphocytes than patients with Stage I disease. Similarly, patients with lymph node metastasis had higher TIGIT (p=0.04) and PD-1 (p=0.05) levels on CD16+ and CD56+ lymphocytes, respectively. No other significant associations could be found between immune check receptors and other parameters. Conclusion: Our results suggest TIL in patients with more advanced stages and younger than 45 years old are more likely to express higher levels of immune checkpoint receptors such as LAG-3, TIM-3, CTLA-4, TIGIT and PD-1. Interestingly, no difference could be found in immune checkpoint receptor expressions in TIL between patients with luminal and TN or HER2+, that would justify immunotherapeutical approaches in selected luminal breast cancers in future trials. Table 1. Significant correlations between immune check point receptor expression and demographic and tumor features CD8PD1CD8LAG3CD16CTLA4CD16TIGITCD16TIM3CD56CTLA4CD56TIGITCD56PD1meanpmeanpmeanpmeanpmeanpmeanpmeanpmeanpAge45 (n:13)9.739.699.049.548.548.087.8810.62Ki-67 (cut off %35) All statistical analyses were evaluated using the Mann Whitney U test. Citation Format: Baran Mollavelioglu, Esin Aktas Cetin, Neslihan Cabioglu, Aykhan Abbasov, Semen Onder, Selman Emiroglu, Mustafa Tukenmez, Mahmut Muslumanoglu, Abdullah Igci, Gunnur Deniz, Vahit Ozmen. Differential expression of novel immune checkpoint receptors expressed on tumor-infiltrating lymphocytes (TIL) in patients with early breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P5-04-24.

Over the last decade, miRNA's have gained considerable interest due to the accumulating evidence regarding their relevant regulatory functions on cancer initiation, proliferation and progression. Furthermore, there are data showing that miRNA's can have a predictive role in response to NAC. The aim of this study is to evaluate the association between miRNA expression profile and response to NAC in patients with BC. 21 patients who received NAC consisting of 4 cycles of antracycline and cyclophosphamide (EC), followed by 4 cycles of docetaxel (T) were included in this study. Tumor core biopsies and blood specimens were collected before initiation, before the 5th cycle and after the completion of NAC for pathologic evaluation, miRNA expression and circulating correlative miRNA levels. The miRNA profile consisted of 9 miRNA's that were previously shown to be related to anthracycline and taxane resistance, as well as those associated with various intracellular functions. Associations with miRNA expression and clinical and pathologic variables, including response were evaluated by the Spearmann Correlation test. Response assessment was carried out every couple of cycles by physical examination, and radiologic evaluation after 4 and 8 cycles, respectively. Clinical response to EC was complete regression in 4.76% (n:1), PR in 66.7% (n:14), minimal response in 23.8% (n:5); and progression in 4.76% (n:1); all patients responded to docetaxel with 1 having CR and 20 PR. Pathologic response evaluation revealed CR in 7 patients (33.3%), good PR (defined as more than 50% regression in the viable tumor cells) in 8 (38.1%) and minimal PR in 6 (28.6%). Baseline levels of miRNA expression before initiation of NAC from the lowest to the highest level were as follows: miR10b (mean relative value (MRV): 0.24), miR-200c, miR34a, miR-11, let7a, miR-200c, miR-21, miR-195 and miR-221 (MRV: 132.5). Although each miRNA tested were in positive correlation with each other (p Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-07-08.