Successful Treatment by Mistletoe Viscum Album Extract of a Patient with Recurrent Triple Negative Breast Cancer who declined Chemotherapy

Triple negative breast cancer (TNBC) is an aggressive breast cancer with a high incidence of recurrence and poor prognosis even with the full spectrum of standard oncology treatments. Ever since its introduction about 100 years ago for cancer therapy, mistletoe viscum album extract (VAE) has been widely used in Europe as a popular complementary therapy for many cancers. We present a case of a patient diagnosed with stage 3c (T3N3bM0(i+)) TNBC who declined chemotherapy after a recurrence 2 years later. The patient received subcutaneous, intratumoral as well as intravenous VAE therapy for her recurrence. Her breast masses and lymph node sizes dramatically decreased under ultrasound examination within 2 months. After nearly three years since the tumor resolution, she is currently being managed on weekly subcutaneous VAE as well as intravenous VAE once in 6-8 weeks. She has excellent quality of life, no sign of tumor recurrence or other adverse side effects from her treatments. Though we believe the best outcomes in oncology are by integrating complementary and conventional care together, this case shows that VAE can have standalone effectiveness at least in some cases. Use of VAE to augment conventional treatments should be considered to bolster breast cancer care.

Prognostic Significance of CSF-1R Expression in Early Invasive Breast Cancer

Colony-stimulating factor-1 receptor (CSF-1R) signaling promotes an immune suppressive microenvironment enriched in M2 macrophages. Given that CSF-1R inhibitors are under investigation in clinical trials, including in breast cancer, CSF-1R expression and association with immune biomarkers could identify patients who derive greater benefit from combination with immunotherapies. TIMER2.0 and bc-GenExMiner v4.7 were used to assess the correlation of CSF1R mRNA with immune infiltrates and prognosis. Following a prespecified training–validation approach, an optimized immunohistochemistry assay was applied to assess CSF-1R on carcinoma cells and macrophages on breast cancer tissue microarray series representing 2384 patients, coupled to comprehensive clinicopathological, biomarker, and outcome data. Significant positive correlations were observed between CSF1R mRNA and immune infiltrates. High carcinoma CSF-1R correlated with grade 3 tumors >2 cm, hormone receptor negativity, high Ki67, immune checkpoint biomarkers, and macrophages expressing CSF-1R and CD163. High carcinoma CSF-1R was significantly associated with poor survival in univariate and multivariate analyses. Adverse prognostic associations were retained in ER+ cases regardless of the presence of CD8+ T cells. CSF-1R+ macrophages were not prognostic. High carcinoma CSF-1R is associated with aggressive breast cancer biology and poor prognosis, particularly in ER+ cases, and identifies patients in whom biomarker-directed CSF-1R therapies may yield superior therapeutic responses.

Non-Coding RNAs Associated With Radioresistance in Triple-Negative Breast Cancer

The resistance that Triple-Negative Breast Cancer (TNBC), the most aggressive breast cancer subtype, develops against radiotherapy is a complex phenomenon involving several regulators of cell metabolism and gene expression; understanding it is the only way to overcome it. We focused this review on the contribution of the two leading classes of regulatory non-coding RNAs, microRNAs (miRNAs) and long non-coding RNAs (lncRNAs), against ionizing radiation-based therapies. We found that these regulatory RNAs are mainly associated with DNA damage response, cell death, and cell cycle regulation, although they regulate other processes like cell signaling and metabolism. Several regulatory RNAs regulate multiple pathways simultaneously, such as miR-139-5p, the miR-15 family, and the lncRNA HOTAIR. On the other hand, proteins such as CHK1 and WEE1 are targeted by several regulatory RNAs simultaneously. Interestingly, the study of miRNA/lncRNA/mRNA regulation axes increases, opening new avenues for understanding radioresistance. Many of the miRNAs and lncRNAs that we reviewed here can be used as molecular markers or targeted by upcoming therapeutic options, undoubtedly contributing to a better prognosis for TNBC patients.

699 ATYR2810 an anti-NRP2 monoclonal antibody targets tumor-associated macrophages

BackgroundNeuropilin-2 (NRP2) is a single transmembrane pleiotropic receptor that utilizes co-receptors for signal transduction and is known to impact tumor progression and metastasis.1 Deletion of NRP2 in murine tumor-associated macrophages (TAMs) downregulated several immunosuppressive and tumor-promoting genes and upregulated immune-stimulatory genes in the myeloid compartment.2 However, little is known about the role of NRP2 in human TAMs. We previously reported significant expression of NRP2 on TAMs derived from triple negative breast cancer (TNBC),3 and demonstrated the ability of ATYR2810, a monoclonal anti-NRP2 antibody, to regulate epithelial-mesenchymal transition (EMT) genes, such as the transcription factor ZEB1, and to enhance chemotherapeutic efficacy for aggressive breast cancer.4 Knowing TAMs play an important role in EMT transition and therapy resistance of cancer, and also rely on ZEB1 for their cancer promoting roles such as immune regulation,5 we sought to investigate the effects of ATYR2810 on TAMs.MethodsMDA-MB-231 TAMs were generated from monocytes in the presence or absence of ATYR2810. TAM phenotypes, gene expression and ability to secrete cytokines were assessed by flow cytometry, qRT-PCR and MSD respectively. TAM suppressive-ness was measured in co-culture experiments. T cell proliferation and activation markers were monitored by flow cytometry and cytokine production by MSD.ResultsNRP2 is highly expressed on TAMs, which suppress T cell proliferation, activation and cytokine release. When differentiated in the presence of ATYR2810, a significant decrease in their suppressive capabilities against T cells was observed. Briefly, T cells were more proliferative, active and altered cytokine production when co-cultured with TAMs exposed to ATYR2810 compared to TAMs differentiated in its absence. Interestingly, we observed a significant decrease in ZEB1 gene and protein expression in ATYR2810 treated TAMs compared to non-treated TAMs. ATYR2810 also decreased the suppressive ability of TAMs when present in co-culture experiments.ConclusionsWe show here for the first time that ATYR2810, known to bind NRP2 tumor cells, can also bind and exert effects on human TAMs. Given the intricate relationship between TAMs and tumors, we believe that this novel finding provides additional insight into the mechanism of action of ATYR2810 as a potential immune regulator. We show for the first time that NRP2 has the ability to regulate ZEB1 expression in TAMs; reducing their suppressive nature, pointing to a novel role of NRP2 in TAMs. These findings indicate ATYR2810s potential to be an effective anti-cancer agent through regulation of ZEB1 in both TAMs and tumors.ReferencesCaunt M, Mak J, Liang W-C, Stawicki S, Pan Q, Tong RK, Kowalski J, Ho C, Reslan HB, Ross J, Berry L, Kasman I, Zlot C, Cheng Z, Le Couter J, Filvaroff EH, Plowman G, Peale F, French D, Carano R, Koch AW, Wu Y, Watts RJ, Tessier-Lavigne M, Bagri A. Blocking Neuropilin-2 function inhibits tumor cell metastasis. 2008;13(4):331–342. https://doi.org/10.1016/j.ccr.2008.01.029.Roy S, Bag AK, Dutta S, Polavaram NS, Islam R, Schellenburg S, Banwait J, Guda C, Ran S, Hollingsworth MA, Singh RK, Talmage JE, Muders MH, Batra SK, Datta K. Macrophage-derived neuropilin-2 exhibits novel tumor-promoting functions. Cancer Res 2018;78(19):5600–5617. doi: 10.1158/0008–5472.CAN-18-0562.Tyler S, Ferrer M, Polizzi C, Da Silva R, Eide L, Walwick K, Seikkula M, Burkart C, Paz S, Nangle L. Neuropilin-2 is expressed on immune cells present in the tumor microenvironment, and may contribute to the suppression of immune regulation leading to progression and metastasis of cancer. Keystone Symposia: Tumor Metabolism and the Microenvironment. 2021. https://www.atyrpharma.com/wp-content/uploads/2021/01/Jan-2021-Keystone-Poster_ST_FINAL.pdf.Xu Z, Burkart C, Goel HL, Rahman J, Polizzi C, Seikkula M, Burman L, Mercurio AM, Nangle LA. A domain-specific antibody to NRP2 down-regulated epithelial-mesenchymanl transition genes and enhanced efficacy of stnadar-of-care therapeutics for aggressive breast cancer. American Society for Cancer Research, 2021. https://www.atyrpharma.com/wp-content/uploads/2021/04/2021Mar_AACR-poster_ZX_Final_v2.pdf.Cortés M, Sanchez-Moral L, de Barrios O, Fernández-Aceñero MJ, Martínez-Campanario MC, Esteve-Codina A, Darling DS, Gyórffy B, Lawrence T, Dean, DC, Postigo A. Tumor-associated macrophages (TAMs) depend on ZEB1 for their cancer-promoting roles. EMBO 2017;36(22):3336–3355. doi: 10.15252/embj.201797345. Epub 2017 Oct 16.

Simvastatin induced ferroptosis for triple-negative breast cancer therapy

Triple-negative breast cancer (TNBC), a management of aggressive breast cancer, remains an unmet medical challenge. Although a wave of efforts had spurred to design novel therapeutic method of TNBC, unpredictable prognosis with lacking effective therapeutic targets along with the resistance to apoptosis seriously limited survival benefits. Ferroptosis is a non-apoptotic form of cell death that is induced by excessive lipid peroxidation, which provide an innovative way to combat cancer. Emerging evidence suggests that ferroptosis plays an important role in the treatment of TNBC cells. Herein, a novel ferroptosis nanomedicine was prepared by loading simvastatin (SIM), a ferroptosis drug, into zwitterionic polymer coated magnetic nanoparticles (Fe3O4@PCBMA) to improve the therapeutic effect of TNBC. The as-obtained Fe3O4@PCBMA-SIM nanoparticles demonstrated more cytotoxicity against MDA-MB-231 than MCF-7 due to the higher expression of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR), which demonstrated that statins could effectively kill TNBC. Further experiments showed that SIM could inhibit the expression of HMGCR to downregulate the mevalonate (MVA) pathway and glutathione peroxidase 4 (GPX4), thereby inducing cancer cell ferroptosis. What’s more, PCBMA endows Fe3O4@PCBMA longer blood circulation performance to enhance their accumulation at tumor sites. Given that Fe3O4 have proven for clinical applications by the U.S. Food and Drug Administration (FDA) and SIM could induce cancer cell ferroptosis, the developed Fe3O4@PCBMA-SIM nanosystem would have great potential in clinics for overcoming the drug resistance brought about by apoptotic drugs to cancer cells.

Prominent Prognostic Factors in Aggressive Breast Cancer: A Review

Context: Breast cancer (BC) is the most common cancer in women worldwide. Hereditary susceptibility created by mutations in autosomal dominant genes is responsible for 5 to 10% of all BC cases in women. Recent studies have identified genes associated with increased risk for aggressive BC, providing the basis for better risk management. Evidence Acquisition: The latest information in National Center for Biotechnology Information (NCBI), Google Scholar, ScienceDirect, and Scopus were the main databases for finding articles. A combination of keywords of ‘metastasis’, ‘invasion’, ‘aggressive breast cancer’, ‘prognostic factor’, ‘mutation’, and ‘cancer treatment’ was searched in the databases to identify related articles. Titles and abstracts of the articles were studied to choose the right articles. Results: Mutations in breast cancer type 1 susceptibility protein (BRCA1) and breast cancer type 2 susceptibility protein (BRCA2) genes are two central players related to the high risk of BC. Mutation in tumor protein p53 (TP53) is another important mutation that leads to triple-negative BC. Although the majority of BC types are not associated with high-throughput mutant genes such as BRCA1, BRCA2, and TP53, they are associated with low-throughput genes, including DNA repair protein Rad50 (RAD50), Nijmegen breakage syndrome gene (NBS1), checkpoint kinase 2 (CHEK2), BRCA1-interacting protein 1 (BRIP1), E-cadherin gene (CDH1) and PALB2, UCHL1, aldehydedehydrogenase1A3 (ALDH1A3), androgen receptor (AR), 5-bisphosphate 3-kinase (PIK3CA), phosphatidylinositol-4, and luminal gene expression that are generally mutated in the global population. High tumor mutational burden (TMB) was associated with improved progression-free survival. Conclusions: The lymph node status, early tumor size, ER, PR, human epidermal growth factor receptor-2 (HER2), and Ki-67 are conventional prognostic factors for BC. However, these factors cannot exactly predict the aggressive behavior of BC. Hence, in this review, we discussed new prognostic factors of aggressive BCs that are useful for the treatment of patients with BC.

Cyclin D1b induces changes in the macrophage phenotype resulting in promotion of tumor metastasis

In breast cancer, tumor-associated macrophages with activated phenotypes promote tumor invasion and metastasis. The more aggressive mesenchymal-like breast cancer cells have a selective advantage, skewing macrophages toward the more immunosuppressive subtype. However, the mechanism underlying this shift is poorly understood. Cyclin D1b is a highly oncogenic variant of cyclin D1. Our previous study showed that non-metastatic epithelial-like breast cancer cells were highly metastatic in vivo when cyclin D1b was overexpressed. The present study determined whether cyclin D1b contributed to the interaction between breast cancer cells and macrophages. The results showed that cyclin D1b promoted the invasion of breast cancer cells in vitro. Specifically, through overexpression of cyclin D1b, breast cancer cells regulated the differentiation of macrophages into a more immunosuppressive M2 phenotype. Notably, tumor cells overexpressing cyclin D1b activated macrophages and induced migration of breast cancer cells. Further investigations indicated that SDF-1 mediated macrophage activation through breast cancer cells overexpressing cyclin D1b. These results revealed a previously unknown link between aggressive breast cancer cells and Tumor-associated macrophages, and highlighted the importance of cyclin D1b activity in the breast cancer microenvironment.

Counterbalance of Foxp3 and IDO expression at different tumor stages in aggressive breast cancer subtypes

Foxp3 and IDO1 are known immunomodulatory molecules involved in tumor escape and could be related to tumor infiltrating lymphocytes (TILs) in the tumor microenvironment. In this study, tumoral Foxp3 and IDO1 expression in breast cancer were evaluated in relation to lymphocyte biomarkers such as CD8 and CD45R0, regulatory T cells, as well as intratumoral and stromal TILs (iTILs and sTILs, respectively). Clinical and histopathological features were also included in the analysis. Foxp3 and IDO1 were found in tumor cells showing mainly cytoplasmic patterns in 60% and 62% tumor samples, respectively. TILs were found in 76% of samples; iTILs were detected in 92% of those samples and sTILs in 55%. Foxp3+ TILs were detected only in 12% of TILs+ samples associated with tumoral Foxp3 expression. Tumoral Foxp3 was mainly expressed at lower tumor stages while IDO1 expression was associated with advanced tumor stages; both correlated with CD8+ TILs which were observed in 77% of TILs+ samples. CD45R0+ were observed in 81% of TILs+ samples and correlated with higher tumor stages and poorly differentiated tumors. In ER negative tumors, an inverse correlation between Foxp3 and IDO1 tumoral expression was found in relation to tumor stage. TNBC subtype showed a positive correlation with the presence of iTILs. In silico analysis showed that Foxp3 and coexpressed genes in breast cancer were associated with immune response genes. Foxp3 was found predominantly in Basal and Her2-enriched subtypes in relation to Luminal A subtype, by RNA seq and RNA microarray database analysis. In conclusion, the expression of Foxp3 and IDO1 in tumors at different stages suggests a potential compensatory mechanism to evade the strong CTL response observed. This is relevant since the cumulative data indicates that Foxp3, as well as IDO1, could be potential targets of immunotherapy in patients with tumors at different stages and for the most aggressive breast cancer subtypes such as TNBC and Her2-enriched.