Mammary Tumor Research Articles

Anti-proliferative and Pro-apoptotic Activity of a Novel Oil-in-Water Nanoemulsion Containing Tea Tree Oil, Quercetin, and Rosmarinic Acid in Breast Cancer Cells

Cancer is one of the main causes of death across major mammalian orders, being the mammary gland tumor the most frequent in females. Surgery is the most widely used prescription for this condition; however, due to its side effects, new and less invasive treatments using bioactive compounds are being sought. Nanostructured systems, such as nanoemulsions (NEs), enable more efficient delivery and protection of these compounds, allowing it to reach its site of action more effectively. The present study aimed to develop oil-in-water (O/W) NEs with tea tree essential oil integrating quercetin, rosmarinic acid, or both and evaluate them as potential therapeutic agents in breast cancer cells. The NEs were characterized by particle size distribution and Z-potential for 4 weeks. Subsequently, their impact on cell viability was assessed in MDA-MB-231 cells, a triple-negative breast cancer cell line. The four NEs had an average size within the nanometer scale (10–9 m), ideal to passive target the anticancer effect (< 200 nm) due to the retention effect and facilitate its permeability. These vehicles were kept stable at 4 °C for 4 weeks. Our findings demonstrate that with initial doses of 3.9 μg/mL of these NEs, significant antiproliferative activity is achieved in cancer cells (~ 80%, p < 0.001); furthermore, the induction of apoptosis through caspase activation was demonstrated in MDA-MB-231 cells. In summary, our preliminary results suggest that nanoemulsions (NEs) of tea tree oil incorporating the bioactive compounds uercetin and rosmarinic acid may have potential therapeutic applications as co-adjuvant treatment of human breast cancer.

Clinical and surgical approaches to mastectomy in dogs: a literature review

Mammary tumors consist of the uncontrolled proliferation of abnormal cells, with hormone-dependent behavior. The frequency of cases involves females exposed to exogenous hormones, less frequently in males. As a form of prevention and control, neutering is suggested. However, given the occurrence of the pathology, the most common treatment is surgery, with excision of the breast parenchyma with a tissue margin, to prevent recurrence. This research aimed to address mammary tumors with a focus on surgical treatment based on topics about the pathology, its relevance and the main surgical techniques for its resolution or control, as there may be recurrence. Among the surgical techniques used, simple, regional, unilateral and bilateral mastectomy stand out, which are indicated according to the patient and the stage of the neoplasia, which is defined by the TNM staging system. The theoretical framework included discussions about its main causes, forms of prevention, and complementary exams to determine the type of tumor so that the surgeon can evaluate the case and determine the best approach to remove the neoformation.

Prognostic Implications of Decorin, E-Cadherin and EGFR Expression in Inflammatory and Non-Inflammatory Canine Mammary Carcinomas.

Inflammatory mammary carcinoma (IMC) is the most aggressive variant of invasive mammary tumours in dogs and in women. Decorin is an extracellular matrix molecule whose expression can be reduced or absent in various human cancers, which is associated with a poor prognosis. E-cadherin is a cell adhesion protein whose expression is reduced in several neoplasms. However, it is overexpressed in inflammatory breast cancers of women. EGFR is also associated with cancer development and is commonly overexpressed in aggressive neoplasms. This study aimed to characterise the expressions of Decorin, E-cadherin, and EGFR in canine inflammatory and non-inflammatory mammary carcinomas (IMC and non-IMC) and to evaluate their expression levels as prognostic indicators for survival and occurrence of metastases. Thirty-three IMC and 43 non-IMC cases were analysed retrospectively and submitted to immunohistochemical analysis. The reactions were quantified in five high-power field images from areas of the highest intensity and frequency of immunostaining (hot spots). We found significantly lower expression of Decorin and higher of E-cadherin and EGFR in canine IMCs. Patients with tumours that exhibited Decorin expression in less than 26.35% of epithelial cells had shorter survival (p = 0.0410) and a higher occurrence of distant metastases (p = 0.0115). E-cadherin is overexpressed in canine IMCs (p < 0.0001), similar to what occurs in women, reinforcing that dogs can be used as a study model for human IMC. EGFR overexpression in canine IMCs (p = 0.0322) provides evidence for potential targeted therapy with tyrosine kinase inhibitors.

Suppression of chemically induced mammary cancer by early-life oral administration of cholera toxin in mice is associated with aberrant regulation of Bmp and Notch signaling pathways.

Lately, significant attention has been drawn towards the potential efficacy of cholera toxin (CT)-an exotoxin produced by the small intestine pathogenic bacterium Vibrio cholera-in modulating cancer-promoting events. In a recent study, we demonstrated that early-life oral administration of non-pathogenic doses of CT in mice suppressed chemically-induced carcinogenesis in tissues distantly located from the gut. In the mammary gland, CT pretreatment was shown to reduce tumor multiplicity, increase apoptosis and alter the expression of several cancer-related molecules. In the present work we investigated the protumorigenic mammary microenvironment for possible associations between early life CT administration and the expression of key components of the Bmp and Notch signaling pathways. Total RNA from mammary tissue samples were retrieved from a recent experiment where FVB/N female mice were preconditioned with CT and later treated with the carcinogen 7,12-dimethylbenzanthracene (DMBA). Real-time PCR was used for relative quantification of gene expression. Our results revealed that CT anti-tumor effects significantly correlated with deregulation of crucial BMP pathway elements, with downregulation of Bmp7 ligand and upregulation of inhibitory Smad6 being the most prominent alterations observed. Concerning Notch signaling pathway, significantly elevated gene expression levels in the CT-treated DMBA mice, as compared to their non-treated counterparts, were also identified at the ligand-receptor level. These findings suggest that CT tumor protective effects in the mammary gland are associated with discerning deregulation of components of both Bmp and Notch signaling pathways and provide insights into the mechanisms underlying CT's anti-cancer outcome.

β-Glucuronidase-Responsive Albumin-Binding Prodrug of Colchicine-Site Binders for Selective cancer Therapy.

The development of novel therapeutic strategies enabling the selective destruction of tumors while sparing healthy tissues is of great interest to improve the efficacy of cancer chemotherapy. In this context, we designed a β-glucuronidase-responsive albumin-binding prodrug programmed to release a potent Isocombretastatin A-4 analog within the tumor microenvironment. When injected at a non-toxic dose in mice bearing orthotopic triple-negative mammary tumors, this prodrug produced a significant anticancer activity, therefore offering a valuable alternative to the systemic administration of the parent drug.

Tumor-Infiltrating Lymphocytes and Neutrophils and Their Location in Canine Mammary Neoplasms with a Solid Arrangement: A Prognostic Factor?

In canine mammary neoplasms, greater inflammation is associated with higher histological grade, lymphatic invasion, and metastases. This retrospective study assessed the density of peri- and intratumoral tumor-infiltrating lymphocytes (TILs), tumor-associated neutrophils (TANs), and CD3+ and CD79+ lymphocytes in canine mammary neoplasms with a solid arrangement, and associated such data with histological types, immunophenotype, prognostic factors, cyclooxygenase-2 (Cox-2) expression and overall and cancer-specific survival. Sixty-one neoplasms with a solid arrangement were classified as malignant myoepitheliomas (6/9.8%), solid papillary carcinomas (8/13.1%), carcinomas with a solid pattern (9/14.8%), basaloid carcinomas (BC) (19/31.1%), and malignant adenomyoepitheliomas (19/31.1%). Intra- and peritumoral TILs, TANs, and TCD3+ and BCD79+ lymphocytes were counted, and based on the resulting median, the neoplasms were divided into low or high cell infiltration. BCs had the lowest density of intratumoral TILs (p = 0.02), and luminal B neoplasms showed a significantly higher density of intratumoral TCD3+ than luminal A cases. Neoplasms with a higher density of peritumoral CD3+ and CD79+ had significantly greater proliferative activity. High infiltration of intratumoral BCD79+ lymphocytes was related to nodal metastasis (p = 0.03). Intratumoral TILs and TCD3+ were associated with shorter survival time. Therefore, intratumoral lymphocyte infiltration is possibly an important feature in the progression of cancer and influences the survival in bitches with solid arrangement neoplasms.

Inhibition of mTOR attenuates the initiation and progression of BRCA1-associated mammary tumors.

Inhibition of mTOR attenuates the initiation and progression of BRCA1-associated mammary tumors.

Possible relationship between long-term post neutering complications in dogs and caregiver burden in the owners

Gonadectomy, commonly known as neutering, is widely used to address dog overpopulation and reduce reproductive disease risks, such as mammary cancer and pyometra. It is also advocated for behavior modification. However, neutering is associated with an increased risk of certain musculoskeletal disorders, obesity, several types of cancer, immune-mediated diseases, and cognitive dysfunction. These conditions may necessitate ongoing care and treatment, that require special care that the caregiver must provide furthermore burdens the caregiver with daily tasks, which encompasses the emotional, physical, social, and financial impact on pet owners caring for chronically ill animals. This burden can result in burnout, health issues, depression, social isolation, and financial stress. The potential benefits and risk of gonadectomy can affect the quality of life of both humans and pets. Relevant research findings should therefore be incorporated into each pet's and owner's particular situation. The purpose of this review is to address the long-term neutering risks and the potential caregiver burden that arises from these risks.

A new twist on superantigen-activated autoimmune disease.

Superantigen-induced (Sag-induced) autoimmunity has been proposed as a mechanism for many human disorders, without a clear understanding of the potential triggers. In this issue of the JCI, McCarthy and colleagues used the SKG mouse model of rheumatoid arthritis to characterize the role of Sag activity in inflammatory arthritis by profiling arthritogenic naive CD4+ T cells. Within the diseased joints, they found a marked enrichment of T cell receptor-variable β (TCR-Vβ) subsets that were reactive to the endogenously encoded mouse mammary tumor virus (MMTV) Sag. Arthritis was improved using reverse transcriptase inhibitors. Moreover, depletion of MMTV Sag-activated TCR-Vβ subsets affected the ability of transferred activated CD4+ T cells to induce disease in mice with severe combined immunodeficiency (SCID). Further virological studies should determine whether endogenous or exogenous MMTV is necessary or sufficient to trigger inflammatory arthritis in the SKG model.

Neuroinflammation mediates the progression of neonate hypoxia-ischemia brain damage to Alzheimer's disease: a bioinformatics and experimental study.

Traumatic brain injury (TBI) can generally be divided into focal damage and diffuse damage, and neonate Hypoxia-Ischemia Brain Damage (nHIBD) is one of the causes of diffuse damage. Patients with nHIBD are at an increased risk of developing Alzheimer's disease (AD). However, the shared pathogenesis of patients affected with both neurological disorders has not been fully elucidated. We here aim to identify the shared molecular signatures between nHIBD and AD. We used an integrated analysis of the cortex gene expression data, targeting differential expression of genes related to the mechanisms of neurodegeneration and cognitive impairment following traumatic brain injury. The gene expression profiles of Alzheimer's disease (GSE203206) and that of Neonate Hypoxia-Ischemia Brain Damage (GSE23317) were obtained from the Gene Expression Omnibus (GEO) database. After identifying the common differentially expressed genes (DEGs) of Alzheimer's disease and neonate Hypoxia-Ischemia Brain Damage by limma package analysis, five kinds of analyses were performed on them, namely Gene Ontology (GO) and pathway enrichment analysis, protein-protein interaction network, DEG-transcription factor interactions and DEG-microRNA interactions, protein-drug interactions and protein-disease association analysis, and gene-inflammation association analysis and protein-inflammation association analysis. In total, 12 common DEGs were identified including HSPB1, VIM, MVD, TUBB4A, AACS, ANXA6, DIRAS2, RPH3A, CEND1, KALM, THOP1, AREL1. We also identified 11 hub proteins, three central regulatory transcription factors, and three microRNAs encoded by the DEGs. Protein-drug interaction analysis showed that CYC1 and UQCRFS1 are associated with different drugs. Gene-disease association analysis shows Mammary Neoplasms, Neoplasm Metastasis, Schizophrenia, and Brain Ischemia diseases are the most relevant to the hub proteins we identified. Gene-inflammation association analysis shows that the hub gene AREL1 is related to inflammatory response, while the protein-inflammation association analysis shows that the hub proteins AKT1 and MAPK14 are related to inflammatory response. This study provides new insights into the shared molecular mechanisms between AD and nHIBD. These common pathways and hub genes could potentially be used to design therapeutic interventions, reducing the likelihood of Alzheimer's disease development in survivors of neonatal Hypoxic-Ischemia brain injury.

Novel combination therapy targeting oncogenic signaling kinase P21 activated Kinase-1 and chemotherapeutic drugs against triple negative breast cancer.

Most of the triple negative phenotype or basal-like molecular subtypes of breast cancers are associated with aggressive clinical behaviour and show poor disease prognosis. Current treatment options are constrained, emphasizing the need for novel combinatorial therapies for this particular tumor subtype. Our group has demonstrated that functionally active p21 activated kinase 1 (PAK1) exhibits significantly higher expression levels in clinical triple negative breast cancer (TNBC) samples compared to other subtypes, as well as adjacent normal tissues. Low PAK1 expression in TNBC was significantly linked to better prognosis, with improved overall survival (OS, p=0.00236) and relapse-free survival (RFS, p=0.0314), as shown by GOBO analysis. To confirm the role of PAK1 as a therapeutic target and to discover novel synergistic chemotherapy drug combinations, we conducted a drug combination screen using triple negative breast cancer cell lines and a mouse metastatic tumor cell line. We identified the combined inhibition of PAK1 inhibitor, NVS-PAK1 with doxorubicin/paclitaxel/methotrexate as a synergistic novel therapeutic approach for treating metastatic TNBC to improve overall survival. This study also indicated a reduction in the effective dosage of the chemotherapeutic drug when combined with NVS-PAK1. Our study demonstrates that combining NVS-PAK1 with each of the chemotherapeutic drugs' doxorubicin, paclitaxel, and methotrexate resulted in decreased colony formation, reduced wound healing capability, and diminished migratory and invasive potential in both TNBC cell lines and 4T1 in vitro. These findings were further validated in orthotopic mouse mammary tumors, confirming that simultaneous PAK1 inhibition alongside chemotherapy significantly enhanced anti-tumor efficacy and reduced metastasis.

Indonesian Newcastle Disease Virus Field Isolate Reduces c-Jun Expression in Rat Mammary Cancer Models

c-Jun is often found to be overexpressed in various cancers, so this gene might be a target for cancer therapy. Newcastle disease virus (NDV) is recognized for its oncolytic properties and potential as a cancer virotherapy agent, with various mechanisms reported to trigger cancer cell death. This study aimed to assess the c-Jun expression in rat mammary cancer models. Rat mammary cancer models were categorized into two treatment groups: the control group (C) and the virotherapy group (V). Group C was administered with 0.5 cc of sterile PBS, while group V received 7 log 2 HAU per 0.5 cc of the Indonesian NDV field isolate Tabanan-1/ARP/2017 intratumorally. The treatment was carried out for four days in a row. Two weeks after treatment, all rats were humanely euthanized, and mammary cancer tissues were excised for further examination. Mammary cancer tissues were examined histopathologically and analyzed using immunohistochemistry to determine intranuclear c-Jun expression, quantified by the H-Score. The results demonstrated that NDV significantly reduced c-Jun expression. It can be inferred that NDV Tabanan-1/ARP/2017 holds potential as a mammary cancer therapy agent by reducing c-Jun expression. This finding is considered novel, as there have been no previous reports of decreased c-Jun expression following virotherapy with NDV.

Targeting BMP-1 enhances anti-tumoral effects of doxorubicin in metastatic mammary cancer: common and distinct features of TGF-β inhibition.

Mammary carcinoma is comprised heterogeneous groups of cells with different metastatic potential. 4T1 mammary carcinoma cells metastasized to heart (4THM), liver (4TLM) and brain (4TBM) and demonstrate cancer-stem cell phenotype. Using these cancer cells we found thatTGF-β is the top upstream regulator of metastatic process. In addition, secretion of bone morphogenetic protein 1 (BMP-1), which is crucial for the proteolytic release of TGF-β, was markedly high in metastatic mammary cancer cells compared to non-metastatic cells. Although TGF-β inhibitors are in clinical trials, systemic inhibition of TGF-β may produce heavy side effects. We here hypothesize that inhibition of BMP-1 proteolytic activity inhibits TGF-β activity and induces anti-tumoral effects. Effects of specific BMP-1 inhibitor on liver and brain metastatic murine mammary cancer cells (4TLM and 4TBM), as well as on human mammary cancer MDA-MB-231 and MCF-7 cells, were examined and compared with the results of TGF-β inhibition. Inhibition of BMP-1 activity markedly suppressed proliferation of cancer cells and enhanced anti-tumoral effects of doxorubicin. Inhibition of BMP-1 activity but not of TGF-β activity decreased colony and spheroid formation. Differential effects of BMP-1 and TGF-β inhibitors on TGF-β secretion was also observed. These results demonstrated for the first time that the inhibition of BMP-1 activity has therapeutic potential for treatment of metastatic mammary cancer and enhances the anti-tumoral effects of doxorubicin.

CD4+ T helper 2 cell-macrophage crosstalk induces IL-24-mediated breast cancer suppression.

CD4+ T cells contribute to antitumor immunity and are implicated in the efficacy of cancer immunotherapies. In particular, CD4+ T helper 2 (Th2) cells were recently found to block spontaneous breast carcinogenesis. However, the antitumor potential of Th2 cells in targeting established breast cancer remains uncertain. Herein, we demonstrate that Th2 cells induced by the topical calcipotriol/thymic stromal lymphopoietin cytokine axis suppressed the growth of established mammary tumors in mice. Interleukin-24 (IL-24), an anticancer cytokine, was highly upregulated in macrophages infiltrating calcipotriol-treated mammary tumors. Macrophages expressed IL-24 in response to IL-4 signaling in combination with Toll-like receptor 4 (TLR4) agonists (e.g., HMGB1) in vitro. Calcipotriol treatment significantly increased HMGB1 release by tumor cells in vivo. CD4+ T cell depletion reduced HMGB1 and IL-24 expression, reversing calcipotriol's therapeutic efficacy. Macrophage depletion and TLR4 inhibition also reduced the therapeutic efficacy of calcipotriol. Importantly, calcipotriol treatment failed to control mammary tumors lacking the IL-24 receptor on tumor cells. Collectively, our findings reveal that Th2 cell-macrophage crosstalk leads to IL-24-mediated tumor cell death, highlighting a promising therapeutic strategy to tackle breast cancer.

Morphometric Parameters as the New Differentiation Criteria of Histological Types of Mammary Gland Tumour in Dogs

Morphometric Parameters as the New Differentiation Criteria of Histological Types of Mammary Gland Tumour in Dogs

Ultrasound-Guided Histotripsy Triggers the Release of Tumor-Associated Antigens from Breast Cancers.

Background/Objectives: There is increasing evidence to indicate that histotripsy treatment can enhance the host anti-tumor immune responses both locally at the targeting tumor site as well as systemically from abscopal effects. Histotripsy is a non-invasive ultrasound ablation technology that mechanically disrupts target tissue via cavitation. A key factor contributing to histotripsy-induced abscopal effects is believed to be the release of tumor-specific antigens (TSAs) or tumor-associated antigens (TAAs) that induce a systemic immune response. In this study, we studied the effect of histotripsy treatment on the release of HER2, a well-defined TAA target for cancer immunotherapy. Methods: A range of doses of histotripsy administered to HER2-postive mammary tumor cells in an in vitro cell culture system and an ex vivo tumor were applied. In addition, a single dose of histotripsy was used for an in vivo murine tumor model. The released proteins, and specifically HER2, in both tumor cell-free supernatants and tumor cell pellets were analyzed by a BCA protein assay, an ultra-performance liquid chromatography (UPLC) assay, and Western blot. Results: Our results showed that histotripsy could significantly trigger the release of HER2 proteins in the current study. The level of HER2 proteins was actually higher in tumor cell-free supernatants than in tumor cell pellets, suggesting that HER2 was released from the intracellular domain into the extracellular compartment. Furthermore, proportionally more HER2 protein was released at higher histotripsy doses, indicating free HER2 was histotripsy-dose-dependent. Conclusions: In conclusion, we have qualitatively and quantitatively demonstrated that histotripsy treatment triggers the release of HER2 from the tumor cells into the extracellular compartment. The histotripsy-mediated release of HER2 antigens provides important insights into the mechanism underlying its immunostimulation and suggests the potential of TSA/TAA-based immunotherapies in numerous cancer types.

Analysis of genetic polymorphisms in the intron 9–10/exon 10 region of the BRCA1 gene in a population sample of dogs with mammary cancer from Uruguay

This study involved clinical and genetic analysis of 15 female dogs with mammary tumors. Fourteen healthy female dogs were used as controls, and blood samples were collected from them for genetic analysis. Polymorphisms located in a splicing region of the largest exon of the BRCA1 gene were studied, both at the population and evolutionary level, in a population of female dogs with different histopathological types of mammary tumors. In the intron 9–10/exon 10 initiation region, two SNP–type polymorphisms are described: SNP1 and SNP2. The SNP1 produces a non–synonymous change with unknown effect on the coding protein. Selected animals underwent surgery, and samples were sent for histopathological analysis. Peripheral blood was also collected for DNA extraction. A region corresponding to intron 9–10/exon 10 of the BRCA1 gene (ENSCAFE00845051080) was amplified by endpoint PCR, with PCR results subsequently confirmed through agarose gel electrophoresis at 1%. PCR products were sequenced to study the polymorphisms identified within this region. No statistically significant differences were observed between the genotype frequencies in both populations (Chi2 0.33, P&gt;0.5), indicating that SNP1 is not linked to mammary tumors in the studied animals. Regarding SNP2, the mutation was not identified in the studied groups (females with mammary tumors and controls), being monomorphic. Although this SNP2 is described in the Ensembl database, there are no genotyping data in reference populations. The phylogenetic analysis of the amplified intron 9–10/exon 10 revealed an evolutionary homology with Canis lupus familiaris, and a more distant relationship with other genera such as Vulpes and Nyctereutes within the Canidae family. It can be concluded that mutations in this splicing region of the largest exon of BRCA1 are not associated with the development of mammary tumors in canines within this group of animals.

Canine mammary tumors as a promising adjunct preclinical model for human breast cancer research: similarities, opportunities, and challenges.

Despite significant progress in the field of human breast cancer research and treatment, there is a consistent increase in the incidence rate of 0.5 percent annually, posing challenges in the development of effective novel therapeutic strategies. The failure rate of drugs in clinical trials stands at approximately 95%, primarily attributed to the limitations and lack of reliability of existing preclinical models, such as mice, which do not mimic human tumor biology. This article examines the potential utility of canine mammary tumors as an adjunct preclinical model for investigating human breast cancer. Given the numerous similarities between canine and human breast cancer, canines present a promising alternative model. The discussion delves into the intricate molecular and clinical aspects of human breast cancer and canine mammary tumors, shedding light on the tumors' molecular profiles, identifying specific molecular markers, and the application of radiological imaging modalities. Furthermore, the manuscript addresses the current constraints of preclinical cancer studies, the benefits of using canines as models, and the obstacles linked to the canine mammary tumors model. By concentrating on these elements, this review aims to highlight the viability of canine models in enhancing our understanding and management of human breast cancer.

Epidemiological analysis of mammary tumors in female dogs in Japan: a study based on Kyushu-Okinawa region.

Canine mammary tumors (CMTs) are common tumors in female dogs (FDs), and at least nearly half of these lesions of malignant. We examined the epidemiology of CMTs in Japan using excisional biopsy cases (n = 7,802) collected from 2005 to 2023 in the Kyushu-Okinawa region. We investigated the prevalence, effects of breed, neutering, and age on CMT and malignant CMT (mCMT) risk through general statistics and multivariate analyses. The distribution of CMT histological types was also compared among different breeds and mixed breeds. In the Cohort (n = 6,197) consisting of cases from primary veterinary hospitals, the numbers of CMT and mCMT cases (2,928 and 822 cases, respectively) and the adjusted prevalence is ranged 4.76-8.09 per 1,000 dogs and increasing over time (P < 0.001). A multivariate model identified breeds with high or low risks of CMT or mCMT. Neutered FDs had lower risk of CMT than intact FDs (risk ratio = 0.57, 95%CI: 0.53-0.61). Compared to the age with the highest incidence, those aged ≥8 and ≥14 years had comparable rates of CMT and mCMT, respectively. Certain breeds exhibited biases regarding CMT histological types compared to mixed breeds. This first epidemiological analysis of CMT in Japan will be a valuable resource for CMT control.

From humans to canines: Unraveling the impact of metabolic health on mammary cancer across species

From humans to canines: Unraveling the impact of metabolic health on mammary cancer across species