bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P4-02-04: Tumor-infiltrating lymphocytes in invasive lobular breast cancer identify a poor prognostic sub-group
Authors: Tille J-C , Saint Martin C , Fuhrmann L , De Koning L , Reyal F , Piccart M , Kirova Y , Cottu P , Carton M , Vincent Salomon A 
Published: May 2019

Abstract
Background:
Tumor infiltrating lymphocytes (TILs) are associated with an improved outcome in triple negative and HER2 breast cancers. Conversely, a recent pioneer study showed that TILs in infiltrating lobular carcinoma (ILC) were associated with a worse prognosis. We aimed at assessing the prognostic impact of TILs in a large cohort of primary surgically treated ILC with a long follow-up in a single institution.
Methods:
We retrospectively reviewed 459 ILC treated at the Curie Institute between 2005 and 2008. Clinico-pathological patients' characteristics were collected and all archived tumor slides were centrally reviewed for TILs quantification. We analyzed stromal TILs scored as a continuous variable. Cox analyses were performed for relapse free survival (RFS) and for overall survival (OS), and chi-square and Fisher test were performed to evaluate the correlation between TILs and other clinico-pathological variables. For statistical analyses, presence or absence of TILs was considered.
Results:
Patients had a mean age of 60.3 years and a median follow-up of 8.8 years, during which 74 local and/or distant relapse events occurred, among which 37 deaths due to the disease. The presence of TILs were significantly associated with greater tumor size (pT), positive nodal status (pN), molecular class (HER2 amplified), and specific morphological nuclear features such as high nuclear grade, multinucleation, macronucleoli and high Nottingham Prognostic Index class. Presence of intra-tumoral lymphocytes is significantly associated with a worse RFS (HR: 2.07 p=0.003) and OS (HR: 4.58 p&lt;0.001) in a multivariate analysis, independently of chemotherapy.
Relapse Free Survival TILs% (IC 95%)% (IC 95%)No95.3 (92.5-98.2)89.0 (84.3-94.0)Yes89.3 (85.1-93.6)74.0 (67.5-81-1)
Overall Survival OS at 5 yearsOS at 10 yearsTILs% (IC 95%)% (IC 95%)No99.5 (97.8-100)95.8 (92.3-99.4)Yes95.3 (92.3-98.1)83.5 (77.8-89.7)
Conclusion:
TILs are an independent prognostic factor associated with a poor outcome in ILC that are easy to assess on diagnostic biopsies. In order to elucidate the role of TILs in ILC, we will further characterize the immune infiltrate and integrate these results with DNA, RNA and protein analyses of the same carcinomas, in order to gain insight into their molecular features and to facilitate the identification of new therapeutic strategies for these ILC with TILs.
Citation Format: Tille J-C, Saint Martin C, Fuhrmann L, De Koning L, Reyal F, Piccart M, Kirova Y, Cottu P, Carton M, Vincent Salomon A. Tumor-infiltrating lymphocytes in invasive lobular breast cancer identify a poor prognostic sub-group [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 P4-02-04.
Abstract P4-02-14: Digitalizing immunohistochemistry analysis (IHC) with quantitative dot blot analysis (QDB) by absolute quantification of biomarkers in FFPE specimens
Authors: Zhang J , Zhang W , Yu G , Tang F , Lv J 
Published: May 2019

Abstract
Purpose: The feasibility of Quantitative Dot Blot (QDB) method for absolute quantification of diagnostic biomarkers was investigated here using diagnostic antibodies for biomarkers in breast cancer tissues, including 4B5 and EP3 for Her2, MIB1 for Ki67. SP2 for Estrogen Receptor and SP1 for progesterone receptor (PR) in more than 300 FFPE specimens. The specificity and accuracy of this method, and the inter-relationship among biomarkers were explored by the absolute expression levels of these biomarkers.
Methods: FFPE specimens of 332 breast cancer patients were provided by local hospital, tissue slices of 2X15 um was de-paraffined and total protein was extracted for QDB analysis, with each sample measured in triplicate in three indepedent experiments. Purified protein standards were included to convert the measurement into absolute levels. IHC and FISH results from local hospital were used to validate the sensitivity and specificity of QDB method.
Results: The results show high consistency with intra-cv less than 7% and inter-cv less than 14%. With Her2, 99.5% concordance was achieved with IHC method, and 92 to 94% concordance with FISH analysis. Both Her2 level and Ki67 levels were significantly increased in patients at grade III based on Nottingham histologic scores.
Conclusion: The QDB method was used here to achieve digitalization of biomarkersl in breast cancer tissue with existing diagnostic antibodies for IHC analysis. The method is simple, objective, consistent, comprehensive and suitable for high throughput analysis. We expect this method to be extended to other diagnostic IHCs with existing diagnostic antibodies, and the easy adoption of this method in clinical practice should have instant impact on pathological practice.
Citation Format: Zhang J, Zhang W, Yu G, Tang F, Lv J. Digitalizing immunohistochemistry analysis (IHC) with quantitative dot blot analysis (QDB) by absolute quantification of biomarkers in FFPE specimens [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 P4-02-14.
Abstract P4-04-12: TZAP mutation leads to poor prognosis of patients with breast cancer
Authors: Lee J-H 
Published: May 2019

Abstract
The zinc finger protein ZBTB48 is a telomere-associated factor that has been renamed as telomeric zinc finger-associated protein (TZAP). It binds preferentially to long telomeres, competing with telomeric repeat factors 1 and 2. However, its expression in cancers has not been investigated. In the present study, we analyzed the TZAP mutation in 128 breast carcinomas (BCs). In addition, its association with telomere length was investigated. The TZAP mutation (c.1272G&gt;A, L424L) was found in 7.8% (10/128) of the BCs and was associated with the N0 stage (p = 0.035). BCs with the TZAP mutation had longer telomeres than those without this mutation (2.62 ± 1.71 vs. 0.98 ± 0.37, p = 0.009). Telomere elongation was shown in 29.7% (38/128) of BCs and was significantly higher in luminal A and triple negative BCs (p &lt; 0.001). Other characteristics were not associated with TZAP mutations and telomere length. Survival analysis showed that the TZAP mutation resulted in a poorer overall survival (52.8 vs. 87.3 months, χ2 = 4.37, P = 0.037). However, telomeres did not have any prognostic value for BCs (P = 0.908). These results suggest that the TZAP mutation is a possible prognostic marker in BC.
Citation Format: Lee J-H. TZAP mutation leads to poor prognosis of patients with breast cancer [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 P4-04-12.
Abstract P4-06-02: Chemoimmunotherapy with cyclophosphamide plus a toll-like receptor 9 (TLR9) agonist eradicates triple negative breast cancer in a murine model
Authors: Campbell MJ , Tandon V , Senman B , Li J , Esserman L 
Published: May 2019

Abstract
Background. Toll-like receptors (TLR) recognize conserved molecular patterns expressed by microbes and together with other molecular sensors serve as a first line of defense, inducing soluble and cellular mediators of innate immunity and initiating key steps of an adaptive immune response. The use of TLR agonists for therapeutic purposes relies on the ability of these compounds to induce, at least partially, some of the immune events that occur during natural infections. For example, synthetic CpG-containing oligodeoxynucleotides (ODN) act as TLR9 agonists, mimicking stimulation of the immune system by bacterial or viral DNA. CpG ODNs are being developed for cancer immunotherapy based on their capacity to stimulate innate and adaptive anti-tumor responses. In this study, we examined the efficacy of intratumorally administered CpG-ODN 1826 alone or in combination with immunomodulatory antibodies (anti-PD-1, anti-OX40, anti-CTLA4) or chemotherapy (cyclophosphamide, paclitaxel) in the syngeneic mouse 4T1 breast tumor model.
Methods. Tumors were implanted orthotopically in the mammary fat pads on the left and right flanks while only one tumor was injected with CpG-ODN 1826. Therapy began 12-14 days post tumor injection when tumors were 8-10 mm in diameter. CpG-ODN 1826 (100 ug) was administered intratumorally, while immunomodulatory antibodies (anti-PD-1 [200 ug], anti-CTLA4 [100 ug], anti-OX40 [400 ug]) and chemotherapies (cyclophosphamide [150 mg/kg], paclitaxel [10 mg/kg]) were administered intraperitoneally. Tumor volume was monitored on both flanks to assess direct and abscopal/systemic anti-tumor activity. Tumor tissue obtained during the treatment regimen was used to evaluate therapy-induced changes in the immune microenvironment.
Results. CpG-ODN 1826, administered intratumorally over 5 consecutive days induced complete regressions in ˜50% of the treated tumors, but only delayed growth in the distant lesions. The immunomodulatory antibodies had little effect on their own and did not add to the therapeutic efficacy of CpG-ODN 1826. The best therapeutic efficacy was obtained with a combination of weekly cyclophosphamide + CpG-ODN 1826 resulting in complete regression of both the CpG injected tumors and the contralateral tumors in 100% of the mice.
Conclusions. These data support the clinical investigation of the combination of a TLR9 agonist (CpG-ODN) with cyclophosphamide in women with breast cancer.
Citation Format: Campbell MJ, Tandon V, Senman B, Li J, Esserman L. Chemoimmunotherapy with cyclophosphamide plus a toll-like receptor 9 (TLR9) agonist eradicates triple negative breast cancer in a murine model [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 P4-06-02.
Abstract P4-06-04: Targeting neo-epitopes from PIK3CA and p53 mutations for immunotherapy of breast cancer
Authors: Chen M , Yuvaraj P , Sharma AA , Narang P , Wilson Sayres MA , Anderson KS 
Published: May 2019

Abstract
Introduction: Preclinical data suggests that priming T cell immunity to mutated or overexpressed proteins can induce tumor rejection, and may potentiate the effects of checkpoint blockade. PIK3CA and p53 are the most commonly mutated genes in breast cancer, and are enriched in high-grade and metastatic tumors. Here, we evaluated the immunogenicity of neo-epitopes derived from common PIK3CA and p53 mutations.
Methods: We applied a custom informatics pipeline, EpitopeHunter, to predict neo-epitopes (8-11mers) from twelve PIK3CA and ten p53 mutations restricted to thirty-nine MHC class I alleles. We selected high affinity neo-epitopes (IEDB score &lt; 500) with predicted poor binding of the matched wild type epitope. We used the predicted peptides to stimulate PBMC from healthy donors in vitro, and measured T cell immunity by interferon-γ ELISPOT. We compared the T cell specificity of mutant vs wild type epitopes. A positive response to an epitope was defined as a response that met both of the following criteria: 1) net number of spots (after subtracting background) was &gt;5 spots/50,000 cells and 2) net number of spots exceeded the background spots plus two SDs.
Results: In total, 1,824 PIK3CA and 1,520 p53 derived peptide sequences were generated. Of these, 42 PIK3CA (2%) and 45 p53 (3%) peptides were predicted to bind to the HLA class I molecules included in our study. Over 90% of the neo-epitopes were 9- and 10-mers, and the predicted neo-epitopes varied across mutations (range 2-10 per mutation). PIK3CAH1047L and p53R248W had the highest number of potential binding neo-epitopes (n=10 each). We successfully generated T cell lines specific for A*0201 PIK3CAH1047L, A*1101 PIK3CAH1047L, A*1101 PIK3CAE542K and A*0201 p53R248W.
Conclusions: Common mutations of PIK3CA and p53 can lead to the generation of potential HLA class I restricted neo-epitopes. We identified four immunogenic neo-epitopes, which may serve as candidates for targeted immunotherapy in breast cancer.
Citation Format: Chen M, Yuvaraj P, Sharma AA, Narang P, Wilson Sayres MA, Anderson KS. Targeting neo-epitopes from PIK3CA and p53 mutations for immunotherapy of breast cancer [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 P4-06-04.
Abstract P4-06-22: Transcription factor T-bet and PD-L1 expression in tumor microenvironment of triple-negative breast cancer
Authors: Mori H , Kubo M , Kai M , Kurata K , Kawaji H , Kaneshiro K , Motoyama Y , Kuroki R , Yamada M , Nishimura R , Okido M , Oda Y , Nakamura M 
Published: May 2019

Abstract
Background: Many analyzes regarding immunotherapies using checkpoint blockade has made it clear that tumor infiltrating lymphocytes (TILs) plays an important role in treating cancers with high levels of somatic mutations such as triple-negative breast cancer (TNBC). We reported the relationship between TILs and PD-L1 expression, and revealed that high-TILs/positive-PD-L1 expression population in TNBC was associated with better prognosis (Oncotarget 2017). However, its molecular mechanism is still unclear. Meanwhile, T-box transcription factor 21 (T-bet) which regulates effecter T-cells activation is derived by stimulation of T-cell receptor and IL-12. Activated T-cells work as antitumor lymphocytes by enhancing the production of cytokines such as INFγ. We focused on T-bet and examined the function of activated T-cells.
Patients and Methods: This study included 242 patients with primary TNBC who underwent resection without neoadjuvant chemotherapy at our three hospitals between January 2004 and December 2014. The immunohistochemistry scoring for CD8 and T-bet expression on TILs was defined as ≥30 per 0.00625mm2. PD-L1 positivity was defined as ≥1% of tumor cells staining positive for PD-L1.
Results: Of the 242 TNBC, CD8 on TILs was expressed as positive in 127 (52.5%) tumors, T-bet on TILs was expressed as positive in 67 (27.7%) tumors, and PD-L1 expression on tumor cells was expressed as positive in 99 (40.9%) tumors. T-bet expression was significantly correlated with CD8 expression (P&lt;0.0001) and PD-L1 expression (P=0.0004). There was no significant difference in recurrence free survival (RFS) and overall survival (OS) regardless of CD8 or PD-L1expression level. Meanwhile, the patients with T-bet-positive tumors had a longer OS, compared to those with T-bet-negative tumors (P = 0.13 in RFS and P = 0.047 in OS). The multivariate analysis revealed that T-bet expression on TILswas an independent and positive prognostic factor for OS(HR = 0.5, 95%CI 0.1-0.9, P = 0.035).
Conclusion: OS was significantly longer among patients with high T-bet expressing TNBC. These results may validate the significance of T-bet as a biomarker for various immunotherapies in TNBC.
Citation Format: Mori H, Kubo M, Kai M, Kurata K, Kawaji H, Kaneshiro K, Motoyama Y, Kuroki R, Yamada M, Nishimura R, Okido M, Oda Y, Nakamura M. Transcription factor T-bet and PD-L1 expression in tumor microenvironment of triple-negative breast cancer [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 P4-06-22.
Abstract P4-06-25: B7-H3 and B7-H4 expression in ductal carcinoma in situ of the breast: Associations with clinicopathologic features and T-cell infiltration
Authors: Lee JS , Kim G-E , Park MH , Yoon JH 
Published: May 2019

Abstract
Background: With the clinical success of immune check point blockades in treating malignant tumors, there are intense investigations in identifying new pathways to activate immune system by targeting immunoregulatory molecules. B7-H3 and B7-H4 play an inhibitory role in T cell function by limiting proliferation and cytokine production. Although several studies have investigated the expression of B7-H3 and B7-H4 in invasive breast cancers, the information of the B7-H3 and B7-H4 molecules in ductal carcinoma in situ (DCIS) remains uncertain.
The present work was undertaken to evaluate the expression of B7-H3 and B7-H4 in DCIS and its association with clinicopathological features in patients with DCIS. In addition, the association of B7-H3 and B7-H4 expression with the T-cell infiltration was also assessed to investigate its roles in the regulation of tumor immune surveillance.
Materials and methods: B7-H3 and B7-H4 expression was examined in 8 pairs of DCIS tissues and matched normal adjacent tissues at mRNA and protein levels by RNAscope in situ hybridization (ISH) and immunohistochemistry. Immunohistochemical staining of B7-H3 and B7-H4 was done in 79 DCIS samples with known hormone receptor (HR) and human epidermal growth factor 2 (HER2) expression using tissue microarray. In addition, immunohistochemical staining was also performed for the T cell lineage markers CD3 and CD8 in DCIS.
Results: RNAscope ISH and immunohistochemistry of B7-H3 and B7-H4 confirmed their increases in DCIS tissues compared with their corresponding normal tissues. B7-H3 and B7-H4 mRNA and protein expression appeared to be concentrated mainly in the DCIS carcinoma cells. High B7-H3 and B7-H4 expression was observed in 58 (73.4%) and 62 (78.5%) cases with DCIS, respectively. High B7-H3 expression was significantly associated with high nuclear grade and presence of comedo-type necrosis (P &lt; 0.05 and P &lt; 0.05, respectively). B7-H3 expression was higher in the HR–/HER2+ subtypes than in the HR+/HER2– subtype (P &lt; 0.05). B7-H3 and B7-H4 expression tended to be associated with low stromal tumor infiltrating lymphocytes density and was negatively related to the CD3+ and CD8+ T cell infiltrates density.
Conclusions: These results suggest that B7-H3 and B7-H4 may play an important role in immune surveillance mechanisms of DCIS and may be useful targets for immune-based therapy to alter or prevent DCIS progression.
Citation Format: Lee JS, Kim G-E, Park MH, Yoon JH. B7-H3 and B7-H4 expression in ductal carcinoma in situ of the breast: Associations with clinicopathologic features and T-cell infiltration [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 P4-06-25.
Abstract P4-06-27: LncRNA NKILA promotes tumour immune evasion by sensitizing tumour-specific t cells to activation-induced cell death
Authors: Song E , Su S , Huang D 
Published: May 2019

Abstract
Activation induced cell death (AICD) of T lymphocytes is critical to maintain T-cell homeostasis, which is adopted by malignant tumors to convey immune evasion by eliminating tumor-reactive cytotoxic T cells. In this study, we demonstrated excessive apoptosis of tumor antigen-specific CTLs in breast and lung cancers. However, the mechanism involved in AICD of tumor-specific T cells remains obscure. Here, we demonstrated that NF-κB activity in tumor-specific T cells is high at the early phase of CTL activation induced by breast tumor antigens, but is suppressed at the later phase. This results in massive apoptosis of tumor-specific CTLs challenged by tumor cells. Interestingly, NKILA, an NFκB interacting lncRNA, sensitizes CTLs to AICD by inhibiting NF-κB activities after their activation, leading to tumor immune evasion. In vivo, administering CTLs with NKILA silencing into immunocompromised mice with breast cancer patient derived xenografts (PDXs) effectively inhibits PDX growth by increasing CTL infiltration. Clinically, NKILA was overexpressed in the tumor specific CTLs of breast and lung cancers, which was associated with less CTL infiltration in the tumors and shorter patient survival. Our findings present the first evidence that AICD in tumor-specific CTLs is crucial to cancer immune evasion, and targeting NKILA in CTLs emerges as a novel anti-tumor immunotherapy.
Citation Format: Song E, Su S, Huang D. LncRNA NKILA promotes tumour immune evasion by sensitizing tumour-specific t cells to activation-induced cell death [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 P4-06-27.
Abstract P4-06-28: Doxorubicin induces cellular senescence in human breast cancer cells and sensitizes them to cytotoxic T-lymphocytes
Authors: Inao T , Iida Y , Kotani H , Harada M 
Published: May 2019

Abstract
[Background]“Cellular senescence” is a state in which cells undergo irreversible cell cycle arrest in response to various cellular stresses. Senescence is induced in not only normal cells but also cancer cells when anti-cancer agents trigger DNA damage. Recent studies have revealed additional feature of senescent cells: increased secretion of various secretory proteins, such as inflammatory cytokines, chemokines, growth factors, and MMPs. This newly recognized senescent phenotype, termed senescence-associated secretory phenotype (SASP), reportedly contribute to tumor recurrence and promotion. Alternatively, senescent cancer cells could be good targets in anti-cancer immunotherapy because they are cell cycle-arrested.
[Objective]In this study, we determined whether ”cellular senescence” could be induced by a chemotherapeutic drug doxorubicin (DXR) and whether senescent cancer cells might increase their susceptibility to cytotoxic T-lymphocytes using human breast cancer cells.
[Methods and Results]A triple-negative (negative for ER, PR, and HER2) human breast cancer cell line (MDA-MB-231) was used. This cell line was treated with DXR for 2 days and examined for their appearance microscopically. The DXR treatment (500 nM) decreased their proliferating ability and increased their cell size. Colony formation assay revealed that cancer cells significantly decreased the number of colonies even with lower doses (3 nM) of DXR. In immunoblot assay, the DXR treatment increased the protein expression of p21, which inhibits cell cycle. In a flow cytometric assay after staining with SPIDER-β-gal, the DXR treatment (500 nM) increased the expression of β-gal in MDA-MB-231 cells. The induction of SA-β-gal in DXR-treated cancer cells were also confirmed by confocal imaging. In addition, the DXR treatment (500 nM) for 2 days beforehand increased their subsequent ability to produce IL-6 and IL-8. Although the DXR treatment (500 nM) decreased the expression of epidermal growth factor receptor (EGFR) on cancer cells, this treatment sensitized them to anti-EGFR chimeric antigen receptor (CAR) T-lymphocytes in apoptosis assay.
[Conclusion]These results suggest that a chemotherapeutic drug DXR can render MDA-MB-231 cells to be senescent and increase their sensitivity to antigen-specific cytotoxic T-lymphocytes. These findings may provide a rationale of combination of chemotherapy and T cell-based anti-cancer immunotherapy.
Citation Format: Inao T, Iida Y, Kotani H, Harada M. Doxorubicin induces cellular senescence in human breast cancer cells and sensitizes them to cytotoxic T-lymphocytes [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 P4-06-28.