Mathematical modelling of tumor-immune interactions in breast cancer.

Immunotherapy Exposes Cancer Stem Cell Resistance and a New Synthetic Lethality

Phenotypic profile of dendritic and T cells in the lymph node of Balb/C mice with breast cancer submitted to dendritic cells immunotherapy

The role of opioids in cancer progression.

Innate immune cells in the tumor microenvironment.

At late stage, nearly all breast cancers are heterogeneous and refractory to treatment, like metaplastic breast cancer is at an early stage. These rare carcinomas are highly aggressive and de-differentiated. They are enriched for mesenchymal and stem cell features and essentially fail current therapies. As metaplastic tumors provide a time-compressed picture of breast cancer progression early on, understanding these tumors will yield insight into mechanisms that drive breast cancer into advanced stages and treatment resistance. To investigate a genetic basis for heterogeneity in metaplastic breast cancer, we established a progression model comprising three cell lines. The cell lines were derived from a primary tumor, a local recurrence and a pleural effusion of a 40-year old patient. The primary tumor was a stage III invasive metaplastic, triple negative, inflammatory breast cancer, resected after neoadjuvant chemotherapy (capecitabine and taxotere, then adriamycin and one cycle of bevacizumab). The local recurrence, biopsied seven months post mastectomy, developed after the patient received adjuvant carboplatin and gemcitabine for 3 cycles and then radiation to the chest wall. At this time, the patient had lung metastases and was treated with taxol and bevacizumab yielding a mixed response. Local invasive growth continued and a malignant pleural effusion developed four months later. Analyzing the genetic and molecular characteristics of this progression model in vitro, its tumorigenicity and metastasis in vivo, and interrogating lead findings in a growing collection of metaplastic tumors helps us to dissect the genetic heterogeneity in breast cancer, and potentially to identify the cell types that drive disease progression and treatment resistance. Our gene expression analyses and genomic evaluations identified epithelial to mesenchymal transition (EMT) as a key characteristic in the progression and treatment resistance of this cancer. Major changes in cytoskeletal genes, chemokines and their receptors, amplification of drug transporter proteins, metalloproteinases and matrix proteins seen with increasing motility and invasiveness along with recruitment of host inflammatory responses in the in vivo model, loss of chromosomal regions harboring known and putative tumor suppressors, and deletions of genes encoding proteins for metabolic inactivation of sex hormones in the breast tissue, along with specific loss of clusters of desmosomal genes are guiding our understanding of metaplastic breast cancer progression. The results provide insight into the development, the extremely invasive nature, and treatment resistance of these tumors. Our collaborative network of clinicians, pathologists, translational genomic researchers and bioinformatics specialists will enable us to identify and prioritize genetic events as disease drivers, prognostic biomarkers of disease progression, and determinants of treatment resistance. Our goal is to identify molecular and functional targets for effective therapy and evaluate them in the clinic. Lessons learned from metaplastic breast cancer will improve our understanding of breast cancer progression in general, and could translate into effective treatments for advanced breast cancer where current standard of care is failing. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD03-07.

The natural cytotoxicity receptors are a unique set of activating proteins expressed mainly on the surface of natural killer (NK) cells. The human natural cytotoxicity receptor family comprises the three type I membrane proteins NKp30, NKp44, and NKp46. Especially NKp30 is critical for the cytotoxicity of NK cells against different targets including tumor, virus-infected, and immature dendritic cells. Although the crystal structure of NKp30 was recently solved (Li, Y., Wang, Q., and Mariuzza, R. A. (2011) J. Exp. Med. 208, 703-714; Joyce, M. G., Tran, P., Zhuravleva, M. A., Jaw, J., Colonna, M., and Sun, P. D. (2011) Proc. Natl. Acad. Sci. U.S.A. 108, 6223-6228), a key question, how NKp30 recognizes several non-related ligands, remains unclear. Therefore, we investigated the parameters that impact ligand recognition of NKp30. Based on various NKp30-hIgG1-Fc fusion proteins, which were optimized for minimal background binding to cellular Fcγ receptors, we identified the flexible stalk region of NKp30 as an important but so far neglected module for ligand recognition and related signaling of the corresponding full-length receptor proteins. Moreover, we found that the ectodomain of NKp30 is N-linked glycosylated at three different sites. Mutational analyses revealed differential binding affinities and signaling capacities of mono-, di-, or triglycosylated NKp30, suggesting that the degree of glycosylation could provide a switch to modulate the ligand binding properties of NKp30 and NK cell cytotoxicity.

SummaryProtein phosphatase 1 regulatory subunit 15A (PPP1R15A) is an important factor in the integrated stress response (ISR) in mammals and may play a crucial role in tumorigenesis. In our studies, we found an inhibitor of PPP1R15A, Sephin1, plays a protumorigenic role in mouse tumor models. By analyzing the single-cell transcriptome data of the mouse tumor models, we found that in C57BL/6 mice, Sephin1 treatment could lead to higher levels of ISR activity and lower levels of antitumor immune activities. Specifically, Sephin1 treatment caused reductions in antitumor immune cell types and lower expression levels of cytotoxicity-related genes. In addition, T cell receptor (TCR) repertoire analysis demonstrated that the clonal expansion of tumor-specific T cells was inhibited by Sephin1. A special TCR + macrophage subtype in tumor was identified to be significantly depleted upon Sephin1 treatment, implying its key antitumor role. These results suggest that PPP1R15A has the potential to be an effective target for tumor therapy.

Tumor Protection Following Vaccination With Low Doses of Lentivirally Transduced DCs Expressing the Self-antigen erbB2

Background: Immunosurveillance suppression, evasion, or avoidance, has emerged as a key targetable hallmark of cancer, driven by e.g. checkpoint expression, T-cell exhaustion, and immunosuppressive tumor microenvironment (TME). Many of these processes generate defined combinations of immune-cell infiltrates at the tumor site, which can be detected by immunohistochemistry (IHC), CyTOF, or more recently can be inferred from gene-expression deconvolution. While significant work has been done to study gene signatures in the TME, the clinical relevance of such immune-cell gene signature on therapy has not been studied to a great extent. We investigate the hypothesis that the individual patterns of immune-cell signatures determine the clinical behavior of breast cancer (BC), in particular response to neoadjuvant chemotherapy. Methods: We performed a retrospective-prospective analysis of a subset of the GeparSepto study (NCT01583426) in which women with primary invasive BC were randomized to either nab-paclitaxel or solvent-based paclitaxel followed by EC. This study was limited to 279 HER2- patients with sufficient quality sample remaining to perform whole-transcriptome RNAseq (~200 × 106 reads per tumor). Immune activity in the TME was inferred by comparing expression of 23 immune-cell-specific gene signatures derived by Bindea et al. (Immunity, 2013) to those from a background population of 1467 similarly-profiled unselected tumor samples from the NantOmics database. Results: Within this cohort the most predominant high immune-cell signatures were for natural killer (NK) cells (71%), and regulatory T-cells (70%). Stimulatory T-cell signatures were high in approximately half of the population including Th2 (53%), effector-memory (53%), follicular helper (51%), Th1 (41%), and Gamma-delta T-cells (39%). While cytotoxic CD8+ T-cell signature was high in only 19.0% of patients, the signature for the CD56dim cytolytic subset of NK cells was high in 48.0% of patients. The most infrequently detected gene signatures were for innate response cell-types: Mast cells (7.5%), Macrophages (10.8%), immature dendritic cells (11.5%) and neutrophils (11.8%). Of the 23 immune signatures, 17 were significantly differentially activated in TNBC compared to hormone-receptor positive (HR+) patients (p<0.05 respectively): 8/10 signatures more active in TNBC are associated with adaptive immune response (e.g. T-cell and B-cell signatures), whereas 5/7 signatures associated with HR+ are related to innate immune response (e.g. eosinophils and dendritic cells). Select adaptive immune signatures were associated with aggressive tumors: Elevated NK CD56dim, Th1, and activated dendritic cell (aDC), signatures were associated with grade 3 tumors as well as with elevated levels of Ki67 (p<0.0001 respectively). The most predictive TME signature for paclitaxel-based therapy was T follicular helper cells (TFH) with DFS and OS hazard ratio of 0.62 (95% CI: 0.47-0.81; p=0.0004) and 0.55 (95% CI: 0.39-0.77; p=0.0005) respectively, as well as a 1.63 (95% CI: 1.12-2.36; p=0.0107) odds-ratio for achieving pCR. Conclusion: Whole-transcriptome sequencing in breast cancer FFPE core biopsies from clinical cohorts can be used to identify immune-cell signatures. Specifically, adaptive immunity through NK rather than T-cell response appears prevalent in high-risk TNBC. The patterns of these immune signatures, in particular the presence of T follicular helper cells, reflect the clinical behavior of breast cancer and might be used to identify tumors with an increased response rate to neoadjuvant chemotherapy. Citation Format: Christopher Szeto, Carsten Denkert, Peter A Fasching, Stephen Benz, Karsten E Weber, Jan Budczies, Andreas Schneeweiss, Elmar Stickeler, Sabine Schmatloch, Christian Jackisch, Thomas Karn, Hans P Sinn, Mathias Warm, Marion van Mackelenbergh, Shahrooz Rabizadeh, Christian Schem, Ernst Heinmöller, Volkmar Müller, Frederik Marmé, Patrick Soon-Shiong, Valentina Nekljudova, Sibylle Loibl, Michael Untch. Landscape of immune-cell signatures in early high-risk breast cancer (BC) reveals clinically-relevant enrichment of immune subpopulations [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 P6-10-04.

10.2217/PME.09.39 © 2009 Future Medicine Ltd “...the antigenic characteristics of tumor cells can be perceived by the innate and cognate immune systems, which represent the extrinsic barriers against tumor progression, or at least a substantial component of the host–tumor equilibrium.” Antoine Tesniere1,2,3 1U848 INSERM, ‘Cancer, Apoptosis & Immunity, Villejuif, France 2Institut Gustave Roussy, Villejuif, France 3Universite Paris Sud, Villejuif, France Guido Kroemer1,2,3 1U848 INSERM, ‘Cancer, Apoptosis & Immunity, Villejuif, France 2Institut Gustave Roussy, Villejuif, France 3Universite Paris Sud, Villejuif, France Thomas Tursz Departement de medecine, Institut Gustave Roussy, Villejuif, France

Chinese‐German Cooperation Group Tumor Immunology: Another inspiring Meeting in Deidesheim

The chemokine system: tuning and shaping by regulation of receptor expression and coupling in polarized responses.

Research Highlights: Immunotherapy

Background: Inflammatory breast cancer (IBC) is a rare but highly aggressive form of the disease but the immune profile associate with pathology and disease progression is poorly characterized. The cells of the innate immune system including natural killer (NK) and dendritic cells (DC) can establish the conditions for an adaptive response to the tumor and can actively kill tumor cells. Alternatively, the inflammatory conditions created by these cells can promote processes like metastasis and invasion. Here we report that immune activation in IBC is associated with a poor prognosis. Methods: A total of 76 breast cancer (BC) patients starting a new line of therapy (22 IBC, 23 metastatic IBC, 8 locally advanced breast cancer, and 23 metastatic) were evaluated for immune phenotypes and cytokine synthesis by DC activated through toll-like receptors (TLR) 7 and 8. Results: Among evaluable patients, 39 (58%) had hormone receptor positive (HR+) primary tumors, 20 (31%) were Her2 positive, and 18 (26%) were triple negative (TN). Among IBC patients, 23 (49%) were HR+, 15 (33%) were Her2+, and 14 (31%) were TN. Among non-IBC patients, 16 (73%) were HR+, 5 (22%) were Her2+, and 4 (17%) were TN. With a mean follow-up time of 7.8 months, 11 (16%) of the patients (10 IBC) died of disease. Most strikingly, BC survivors had a higher median CD4 count than deceased patients (p = .023) and was a significant predictor of outcome (p = 0.01). CD3 and CD19 lymphocytes of BC survivors had higher expression of CXCR4 than deceased patients (p = .038 and p = .029). In contrast, survivors also had lower lymphocyte percentages of all NK sub-types including ADCC NK (p = .008), non-ADCC NK (p = .001), exhausted (p = .014) and non-exhausted NK (p = .005) than deceased patients. Although DC counts were not significantly different, the percentage of TLR-induced pDC producing IFN-α was lower in BC survivors than deceased patients (p = 0.025). Conversely, the percentage of pDC constitutively producing the anti-inflammatory cytokine IL-10 was higher in survivors compared to non-survivors (p = .007). Compared with IBC non-survivors, IBC survivors had a higher % of pDC producing IL-10 (p = 0.005) and fewer mDC producing TNF-α (p = .030) following TLR activation. With respect to antigen presentation, mDC of BC survivors had lower expressions of the co-stimulatory molecules CD40 (p= .036), CD80 (p = .013) and CD86 (p = .006) than mDC of deceased patients. Conclusion: We observed low CD4 counts in IBC that were associated with a poor prognosis. Additionally, immune activation as evidenced by DC pro-inflammatory cytokine production, expression of costimulatory molecules, and increased NK counts was associated with poor prognosis. Together, these data suggest that the innate immune system is highly active in the late stages of disease but the adaptive response is severely compromised by factors still unknown that can contribute to the peculiar clinical presentation and aggressive features of the disease. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5583.

SUMMARYTreatments aiming to augment immune checkpoint blockade (ICB) in cancer often focus on T cell immunity, but innate immune cells may have important roles to play. Here, we demonstrate a single-dose combination treatment (termed AIP) using a pan-tumor-targeting antibody surrogate, half-life-extended interleukin-2 (IL-2), and anti-programmed cell death 1 (PD-1), which primes tumors to respond to subsequent ICB and promotes rejection of large established tumors in mice. Natural killer (NK) cells and macrophages activated by AIP treatment underwent transcriptional reprogramming; rapidly killed cancer cells; governed the recruitment of cross-presenting dendritic cells (DCs) and other leukocytes; and induced normalization of the tumor vasculature, facilitating further immune infiltration. Thus, innate cell-activating therapies can initiate critical steps leading to a self-sustaining cycle of T cell priming driven by ICB.