Evolution of synchronous bilateral breast cancers provide insights into interactions between host, tumor and immunity.

Synchronous bilateral breast cancer (sBBC) occurs after both breasts have been affected by the same germline genetics, reproductive life factors and environmental exposures for decades. It represents an opportunity to decipher the complex interplay between host, tumor, immune system and response to neoadjuvant chemotherapy (NAC). On a cohort of 17575 BCs treated between 2005 and 2012, sBBCs (n=404) were associated with less aggressive proliferative patterns and higher rates of luminal breast cancers (BCs) when compared with unilateral BCs (n=17171). The left and right tumors were concordant for the majority of clinical and pathological features. Tumor pairs of concordant BC subtype were more frequent than pairs of discordant BC subtype, with notably a particularly high frequency of pairs of luminal BCs. Intriguingly, both the levels of tumor infiltrating lymphocytes (TILs) and the response to NAC were modified by the subtype of the contralateral tumors. Whole exome sequencing and RNAseq analyses revealed that left and right tumors were independent from a somatic mutation and transcriptomic point of view, while primary tumors (PT) before NAC and specimens with residual disease (RD) after NAC were more closely related. The analysis of the TCR repertoire identified very little overlap between patients, while common clones were shared in bilateral tumors within each patient. After NAC, the TCR repertoire of RD was enriched and expanded with clones edited by the contralateral PT.

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Molecular Imaging Reveals a High Degree of Cross-Seeding of Spontaneous Metastases in a Novel Mouse Model of Synchronous Bilateral Breast Cancer

Molecular Imaging and Biology ◽

10.1007/s11307-021-01630-z ◽

2021 ◽

Author(s):

Shirley Liu ◽

Nivin N Nyström ◽

John J Kelly ◽

Amanda M Hamilton ◽

Yanghao Fu ◽

...

Keyword(s):

Breast Cancer ◽

Molecular Imaging ◽

Fluorescence Microscopy ◽

Cancer Cells ◽

Ex Vivo ◽

Bilateral Breast Cancer ◽

Short Time Interval ◽

Primary Tumors ◽

Synchronous Bilateral Breast Cancer ◽

High Degree

Abstract Purpose Synchronous bilateral breast cancer (SBBC) patients present with cancer in both breasts at the time of diagnosis or within a short time interval. They show higher rates of metastasis and lower overall survival compared to women with unilateral breast cancer. Here we established the first preclinical SBBC model and used molecular imaging to visualize the patterns of metastasis from each primary tumor. Procedures We engineered human breast cancer cells to express either Akaluc or Antares2 for bioluminescence imaging (BLI) and tdTomato or zsGreen for ex vivo fluorescence microscopy. Both cell populations were implanted into contralateral mammary fat pads of mice (n=10), and dual-BLI was performed weekly for up to day 29 (n=3), 38 (n=4), or 42 (n=3). Primary tumors and lungs were fixed, and ex vivo fluorescence microscopy was used to analyze the cellular makeup of micrometastases. Results Signal from both Antares2 and Akaluc was first detected in the lungs on day 28 and was present in 9 of 10 mice at endpoint. Ex vivo fluorescence microscopy of the lungs revealed that for mice sacrificed on day 38, a significant percentage of micrometastases were composed of cancer cells from both primary tumors (mean 37%; range 27 to 45%), while two mice sacrificed on day 42 showed percentages of 51% and 70%. Conclusions A high degree of metastatic cross-seeding of cancer cells derived from bilateral tumors may contribute to faster metastatic growth and intratumoral heterogeneity. We posit that our work will help understand treatment resistance and optimal planning of SBBC treatment.

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Molecular imaging reveals a high degree of cross-seeding of spontaneous metastases in a novel mouse model of synchronous bilateral breast cancer

10.1101/2020.11.27.401786 ◽

2020 ◽

Author(s):

Shirley Liu ◽

Nivin N Nyström ◽

John J Kelly ◽

Amanda M Hamilton ◽

Yanghao Fu ◽

...

Keyword(s):

Breast Cancer ◽

Molecular Imaging ◽

Fluorescence Microscopy ◽

Cancer Cells ◽

Ex Vivo ◽

Bilateral Breast Cancer ◽

Short Time Interval ◽

Primary Tumors ◽

Synchronous Bilateral Breast Cancer ◽

High Degree

ABSTRACTSynchronous bilateral breast cancer (SBBC) patients present with cancer in both breasts at the time of diagnosis or within a short time interval. They show higher rates of metastasis and lower overall survival compared to women with unilateral breast cancer. However, the lack of a preclinical model has led to a dearth in knowledge regarding the patterns of SBBC metastasis. Here we established an SBBC model and used molecular imaging to visualize the development of spontaneous lung metastases arising from each primary tumor. We engineered human breast cancer cells to express either Akaluc or Antares2 for bioluminescence imaging (BLI), and tdTomato or zsGreen for ex vivo fluorescence microscopy. Both cell populations were implanted into contralateral mammary fat pads of mice (n=10) and BLI was performed weekly for up to day 29 (n=3), 38 (n=4), or 42 (n=3). Signal from both Antares2 and Akaluc was first detected in the lungs on day 28 and was present in 9 of 10 mice at endpoint. Ex vivo fluorescence microscopy of the lungs revealed that for mice sacrificed on day 38, a significant percentage of micrometastases were composed of cancer cells from both primary tumors (mean 37%; range 27% to 45%), while two mice sacrificed on day 42 showed percentages of 51% and 70%. These results reveal a high degree of metastatic cross-seeding which may contribute to faster metastatic growth and intratumoral heterogeneity. We posit our work will help understand treatment resistance and optimal planning of treatment for SBBC patients.

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Identification of shared TCR sequences from T cells in human breast cancer using emulsion RT-PCR

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1606994113 ◽

2016 ◽

Vol 113 (29) ◽

pp. 8272-8277 ◽

Cited By ~ 33

Author(s):

Daniel J. Munson ◽

Colt A. Egelston ◽

Kami E. Chiotti ◽

Zuly E. Parra ◽

Tullia C. Bruno ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

T Cell ◽

Peripheral Blood ◽

Bilateral Breast Cancer ◽

Tumor Infiltrating Lymphocytes ◽

Specific Response ◽

Rt Pcr ◽

Primary Tumors ◽

Infiltrating Lymphocytes

Infiltration of T cells in breast tumors correlates with improved survival of patients with breast cancer, despite relatively few mutations in these tumors. To determine if T-cell specificity can be harnessed to augment immunotherapies of breast cancer, we sought to identify the alpha–beta paired T-cell receptors (TCRs) of tumor-infiltrating lymphocytes shared between multiple patients. Because TCRs function as heterodimeric proteins, we used an emulsion-based RT-PCR assay to link and amplify TCR pairs. Using this assay on engineered T-cell hybridomas, we observed ∼85% accurate pairing fidelity, although TCR recovery frequency varied. When we applied this technique to patient samples, we found that for any given TCR pair, the dominant alpha- or beta-binding partner comprised ∼90% of the total binding partners. Analysis of TCR sequences from primary tumors showed about fourfold more overlap in tumor-involved relative to tumor-free sentinel lymph nodes. Additionally, comparison of sequences from both tumors of a patient with bilateral breast cancer showed 10% overlap. Finally, we identified a panel of unique TCRs shared between patients’ tumors and peripheral blood that were not found in the peripheral blood of controls. These TCRs encoded a range of V, J, and complementarity determining region 3 (CDR3) sequences on the alpha-chain, and displayed restricted V-beta use. The nucleotides encoding these shared TCR CDR3s varied, suggesting immune selection of this response. Harnessing these T cells may provide practical strategies to improve the shared antigen-specific response to breast cancer.

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The number of FoxP3-positive tumor-infiltrating lymphocytes in patients with synchronous bilateral breast cancer

Breast Cancer ◽

10.1007/s12282-020-01049-4 ◽

2020 ◽

Vol 27 (4) ◽

pp. 586-593

Author(s):

Risa Goto ◽

Yuko Hirota ◽

Tomoyuki Aruga ◽

Shinichiro Horiguchi ◽

Sakiko Miura ◽

...

Keyword(s):

Breast Cancer ◽

Bilateral Breast Cancer ◽

Tumor Infiltrating Lymphocytes ◽

Synchronous Bilateral Breast Cancer ◽

Infiltrating Lymphocytes

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Syllabification of the Divine Comedy

Journal on Computing and Cultural Heritage ◽

10.1145/3459011 ◽

2021 ◽

Vol 14 (3) ◽

pp. 1-26

Author(s):

Andrea Asperti ◽

Stefano Dal Bianco

Keyword(s):

Text Processing ◽

Solution Space ◽

Generative Models ◽

Divine Comedy ◽

Point Of View ◽

Computer Assisted ◽

Wide Range ◽

The Divine Comedy ◽

Left And Right ◽

Assisted Analysis

We provide a syllabification algorithm for the Divine Comedy using techniques from probabilistic and constraint programming. We particularly focus on the synalephe , addressed in terms of the "propensity" of a word to take part in a synalephe with adjacent words. We jointly provide an online vocabulary containing, for each word, information about its syllabification, the location of the tonic accent, and the aforementioned synalephe propensity, on the left and right sides. The algorithm is intrinsically nondeterministic, producing different possible syllabifications for each verse, with different likelihoods; metric constraints relative to accents on the 10th, 4th, and 6th syllables are used to further reduce the solution space. The most likely syllabification is hence returned as output. We believe that this work could be a major milestone for a lot of different investigations. From the point of view of digital humanities it opens new perspectives on computer-assisted analysis of digital sources, comprising automated detection of anomalous and problematic cases, metric clustering of verses and their categorization, or more foundational investigations addressing, e.g., the phonetic roles of consonants and vowels. From the point of view of text processing and deep learning, information about syllabification and the location of accents opens a wide range of exciting perspectives, from the possibility of automatic learning syllabification of words and verses to the improvement of generative models, aware of metric issues, and more respectful of the expected musicality.

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Tumor-Infiltrating CD20+ B Lymphocytes: Significance and Prognostic Implications in Oral Cancer Microenvironment

Cancers ◽

10.3390/cancers13030395 ◽

2021 ◽

Vol 13 (3) ◽

pp. 395

Author(s):

Faustino Julián Suárez-Sánchez ◽

Paloma Lequerica-Fernández ◽

Juan Pablo Rodrigo ◽

Francisco Hermida-Prado ◽

Julián Suárez-Canto ◽

...

Keyword(s):

B Lymphocytes ◽

Clinical Stage ◽

Inverse Correlation ◽

Tumor Infiltrating Lymphocytes ◽

Immunohistochemical Analysis ◽

Second Primary ◽

Strong Positive Correlation ◽

Related Factors ◽

Primary Tumors ◽

Sox2 Expression

Immunohistochemical analysis of stromal/tumoral CD20+ B lymphocytes was performed in 125 OSCC patients. Correlations with immune profiles CD4+, CD8+, and FOXP3+ tumor-infiltrating lymphocytes (TILs), tumoral PD-L1, and stem-related factors NANOG and SOX2 were assessed, and also associations with clinical data and patient survival. There was a strong positive correlation between the infiltration of CD20+ B lymphocytes and other immune profiles (i.e., CD4+, CD8+, and FOXP3+ TILs, and CD68+ and CD163+ macrophages) both in stroma and tumor nests. Strikingly, CD20+ TILs were inversely correlated with NANOG/SOX2 expression. Stromal CD20+ TILs were significantly associated with T classification and second primary tumors. A stratified survival analysis showed that tumoral CD20+ TILs were significantly associated with prognosis in male and younger patients, with tobacco or alcohol consumption, high tumoral CD8+ TILs, low tumoral infiltration by CD68+ macrophages, positive PD-L1 expression, and negative NANOG/SOX2. Multivariate Cox analysis further revealed clinical stage and tumoral CD20+ TILs independently associated with disease-specific survival (HR = 2.42, p = 0.003; and HR = 0.57, p = 0.04, respectively). In conclusion, high CD20+ TIL density emerges as an independent good prognostic factor in OSCC, suggesting a role in antitumor immunity. This study also uncovered an inverse correlation between CD20+ TILs and CSC marker expression.

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PD-L1 Expression after Neoadjuvant Chemotherapy in Triple-Negative Breast Cancers Is Associated with Aggressive Residual Disease, Suggesting a Potential for Immunotherapy

Cancers ◽

10.3390/cancers13040746 ◽

2021 ◽

Vol 13 (4) ◽

pp. 746

Author(s):

Beatriz Grandal ◽

Manon Mangiardi-Veltin ◽

Enora Laas ◽

Marick Laé ◽

Didier Meseure ◽

...

Keyword(s):

Neoadjuvant Chemotherapy ◽

Triple Negative ◽

Residual Disease ◽

Checkpoint Inhibitors ◽

Rapid Development ◽

Complete Response ◽

Tumor Burden ◽

Small Subset ◽

Breast Cancers ◽

Residual Cancer Burden

The consequences of neoadjuvant chemotherapy (NAC) for PD-L1 activity in triple-negative breast cancers (TNBC) are not well-understood. This is an important issue as PD-LI might act as a biomarker for immune checkpoint inhibitors’ (ICI) efficacy, at a time where ICI are undergoing rapid development and could be beneficial in patients who do not achieve a pathological complete response. We used immunohistochemistry to assess PD-L1 expression in surgical specimens (E1L3N clone, cutoff for positivity: ≥1%) on both tumor (PD-L1-TC) and immune cells (PD-L1-IC) from a cohort of T1-T3NxM0 TNBCs treated with NAC. PD-L1-TC was detected in 17 cases (19.1%) and PD-L1-IC in 14 cases (15.7%). None of the baseline characteristics of the tumor or the patient were associated with PD-L1 positivity, except for pre-NAC stromal TIL levels, which were higher in post-NAC PD-L1-TC-positive than in negative tumors. PD-L1-TC were significantly associated with a higher residual cancer burden (p = 0.035) and aggressive post-NAC tumor characteristics, whereas PD-L1-IC were not. PD-L1 expression was not associated with relapse-free survival (RFS) (PD-L1-TC, p = 0.25, and PD-L1-IC, p = 0.95) or overall survival (OS) (PD-L1-TC, p = 0.48, and PD-L1-IC, p = 0.58), but high Ki67 levels after NAC were strongly associated with a poor prognosis (RFS, p = 0.0014, and OS, p = 0.001). A small subset of TNBC patients displaying PD-L1 expression in the context of an extensive post-NAC tumor burden could benefit from ICI treatment after standard NAC.

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