Triple-negative Breast Cancer Recurrence Research Articles

19F MRI/CEUS Dual Imaging-Guided Sonodynamic Therapy Enhances Immune Checkpoint Blockade in Triple-Negative Breast Cancer.

Treatment of highly aggressive triple-negative breast cancer (TNBC) in the clinic is challenging. Here, a liposome nanodrug (LP@PFH@HMME) integrating imaging agents and therapeutic agents for bimodal imaging-guided sonodynamic therapy (SDT) is developed, which boosted immunogenicity to enable potent immunotherapy via immune checkpoint blockade (ICB) in TNBC. In the acidic tumor microenvironment (TME), LP@PFH@HMME undergoes "nano-to-micro" transformation due to a pH-responsive lipid fusion, which makes droplets much more sensitive to ultrasound (US) in contrast-enhanced ultrasound (CEUS) and SDT studies. The nanodrug demonstrates robust bimodal imaging ability through fluorine-19 magnetic resonance imaging (19F MRI) and CEUS bimodal imaging, and it exhibits excellent solubility in aqueous solution with relatively high 19F content and desirable long transverse relaxation time (T2 = 1.072 s), making it suitable for high-performance 19F MRI, in addition to effective accumulation of nanodrugs after tail vein injection. Thus, 19F MRI/CEUS dual imaging is achievable to show adequate time points for US irradiation of tumor sites to induce highly effective SDT, which produces abundant reactive oxygen species (ROS) triggering immunogenic cell death (ICD) to assist ICB-based immunotherapy. The combination treatment design of sonodynamic therapy with immunotherapy effectively inhibited TNBC growth and recurrence, highlighting the promise of multifunctional nanodrugs in treating TNBC.

Predicting locoregional recurrence in triple-negative breast cancer using novel molecular subtyping.

e12549 Background: Triple-negative breast cancer (TNBC) presents a poorer prognosis compared to other breast cancer subtypes, characterized by a higher likelihood of early recurrence and increased incidence of visceral metastases. Evidence from numerous randomized clinical trials indicates that postmastectomy radiotherapy (PMRT) reduces locoregional recurrence (LRR) and improves breast cancer survival in TNBC. Previous reports creatively divided TNBC into four molecular subtypes for personalized treatment. The present study aims to elucidate patterns of LRR in different subtypes of TNBC after PMRT. Methods: From 2019 to 2022, data from 349 AJCC 7th edition stage II-III breast cancer patients treated with mastectomy and PMRT were retrospectively analyzed. These patients were classified into the basal-like immune-suppressed (BLIS) and non-BLIS groups based on the immunohistochemistry biomarkers. The cumulative incidence function was applied to estimated LRR rates, with death as a competing risk, using Gray's test for comparisons. Results: According to the IHC results, a total of 255 patients belonged to the non-BLIS group and 94 to the BLIS group. The 3-year LRR for the entire cohort was 5.16%. There were 9 cases each in the non-BLIS subtype (3.5%) and BLIS subtype (9.6%). The cumulative incidence of LRR between the two groups was significantly different, with a rate of 4.0% in the non-BLIS group and 9.0% in the BLIS group ( P = 0.02). Among the 18 patients with LRR in the whole population, 11 had isolated LRR and 7 were diagnosed with LRR with concurrent distant metastases. The most common area of recurrence in both subtypes was the supraclavicular region. The majority of recurrences occurred within the RT field (in-field), and no isolated out-of-field relapse was observed. In multivariate analysis, independent factors predicting higher LRR were the BLIS subtype (HR = 3.06, P = 0.02). Conclusions: Study results suggest that TNBC of the BLIS subtype is at higher risk for LRR. Immunohistochemistry-based molecular typing may serve as a prognostic biomarker to guide postmastectomy radiotherapy in patients with TNBC. New strategies are needed to improve local control rates in patients with BLIS subtype.

Shikonin and chitosan-silver nanoparticles synergize against triple-negative breast cancer through RIPK3-triggered necroptotic immunogenic cell death

Necroptotic immunogenic cell death (ICD) can activate the human immune system to treat the metastasis and recurrence of triple-negative breast cancer (TNBC). However, developing the necroptotic inducer and precisely delivering it to the tumor site is the key issue. Herein, we reported that the combination of shikonin (SHK) and chitosan silver nanoparticles (Chi-Ag NPs) effectively induced ICD by triggering necroptosis in 4T1 cells. Moreover, to address the lack of selectivity of drugs for in vivo application, we developed an MUC1 aptamer-targeted nanocomplex (MUC1@Chi-Ag@CPB@SHK, abbreviated as MUC1@ACS) for co-delivering SHK and Chi-Ag NPs. The accumulation of MUC1@ACS NPs at the tumor site showed a 6.02-fold increase compared to the free drug. Subsequently, upon reaching the tumor site, the acid-responsive release of SHK and Chi-Ag NPs from MUC1@ACS NPs cooperatively induced necroptosis in tumor cells by upregulating the expression of RIPK3, p-RIPK3, and tetrameric MLKL, thereby effectively triggering ICD. The sequential maturation of dendritic cells (DCs) subsequently enhanced the infiltration of CD8+ and CD4+ T cells in tumors, while inhibiting regulatory T cells (Treg cells), resulting in the effective treatment of primary and distal tumor growth and the inhibition of TNBC metastasis. This work highlights the importance of nanoparticles in mediating drug interactions during necroptotic ICD.

Identification of a Gene Expression Signature to Predict the Risk of Early Recurrence and the Degree of Immune Cell Infiltration in Triple-negative Breast Cancer.

Patients with triple-negative breast cancer (TNBC) have a high rate of recurrence within 3 years of diagnosis and a high rate of death within 5 years compared to other subtypes. The number of clinical trials investigating various new agents and combination therapies has recently increased; however, current strategies benefit only a minority of patients. This study aimed to identify specific genes that predict patients at high risk of recurrence and the immune status of the tumor microenvironment at an early stage, thereby providing insight into potential therapeutic targets to improve clinical outcomes in TNBC patients. We evaluated the prognostic significance of microarray mRNA expression of 20,603 genes in 233 TNBC patients from the METABRIC dataset and further validated the results using RNA-seq mRNA expression data in 143 TNBC patients from the GSE96058 dataset. Eighteen differentially expressed genes (AKNA, ARHGAP30, CA9, CD3D, CD3G, CD6, CXCR6, CYSLTR1, DOCK10, ENO1, FLT3LG, IFNG, IL2RB, LPXN, PRKCB, PVRIG, RASSF5, and STAT4) identified in both datasets were found to be reliable biomarkers for predicting TNBC recurrence and progression. Notably, the genes whose low expression was associated with increased risk of recurrence and death were immune-related genes, with significant differences in levels of immune cell infiltration in the tumor microenvironment between high- and low- expression groups. Genes reported herein may be effective biomarkers to identify TNBC patients who will and will not benefit from immunotherapy and may be particularly important genes for developing future treatment strategies, including immunotherapy.

Hyperacute Radiation Pneumonitis after Severe irAE.

A 54-year-old woman presented to an outpatient clinic with a recurrence of triple-negative breast cancer and multiple bone metastases. The patient had a large mass lesion of 10 cm on the sternum. She received the immune checkpoint inhibitors pembrolizumab and taxane. Initially, the patient responded excellently to treatment, but stopped pembrolizumab for grade IV skin toxicity with multiple ulcerative wounds over the bilateral leg and trunk. The lesions abated following administration of antibiotics and oral prednisolone for two months. After that, she was referred to the radiation oncology department for further treatment. She received radiotherapy for the sternum mass but stopped radiation at 42Gy/21 fractions for severe dyspnea and fever. Blood sampling found leukocytosis with neutrophil predominance. Chest radiography showed bilateral lung infiltration. Pulmonary CT scan yielded bilateral lung patchy consolidation compatible with radiation isodose-line. Bronchial lavage showed positive Pneumocystis jiroveci PCR. Dyspnea improved after titrating methylprednisolone within two days. The patient recovered well with TMP-SMX and glucocorticoids after the initiation of therapy.

Triple Negative Breast Cancer (TNBC): Signalling pathways-Role of plant-based inhibitors

This review paper highlights and updates the role of plant based inhibitors in controlling the triple-negative breast cancer (TNBC) via blocking the TNBC signalling pathways. Breast cancer is the most common cancer in women. There are several identified types, among which the triple-negative breast cancer (TNBC) is the most fatal for patients. Conventionally, the triple-negative breast cancer (TNBC) is defined by the lack of estrogen receptor (ER), progesterone receptor (PR), and human epidermal receptor 2 (HER2) in breast cancer cells. It is well-known for its metastatic, aggressive characteristics and poor outcome in the clinic. Till today, the treatment of TNBC patients is still a challenging task due to the absence of appropriate targets for drugs. Despite the successes of emerging targeted therapies, relapse, recurrence, and therapy failure rates in TNBC significantly outpace other subtypes of breast cancer. Mounting evidence suggests an accumulation of therapy resistant, Cancer Stem Cell (CSC) populations within TNBCs contributes to poor clinical outcomes. These Cancer Stem Cells (CSC) are enriched in TNBC compared to non-TNBC breast cancers. Approximately 60% of drugs currently used for cancer treatment have been isolated from natural products. Natural plant based products are a well-known treasure house for the development of novel anticancer drugs. Many plant-derived natural compounds have anti-cancer properties, including berberine quercetin, formononetin, calycosin, polyphenols, bioflavonoids, carotene, vitamins, and andminerals. Many plant-derived natural compounds, including vinka alkaloids, vinblastine and vincristine, luteolin, α-mangostin, piperine, silibinin, apigenin, quercetin, fisetin, resveratrol, genistein, 10-gingerol, chalcones, curcumin, epigallocatechin gallate, cyanidin-3-O-glucoside, and glycyrrhizin, have shown anti-cancer properties, especially in the treatment of TNBCs. Therefore, more human clinical trial data is warranted for further evaluation of plant extracts for the treatment of TNBCs.

Abstract 2420: Evaluating circulating tumor DNA (ctDNA) as a prognostic biomarker utilizing a tissue-free epigenomic assay in early-stage triple negative breast cancer (TNBC)

Abstract Background: ctDNA has shown promise as a prognostic biomarker in early-stage solid tumors. Current breast cancer clinical guidelines do not recommend routine screening for metastatic disease outside of clinical features suggestive of disease recurrence. The detection of ctDNA may be a useful biomarker for the management of early-stage cancer patients to identify patients who are at high risk of recurrence. Methods: Patients with stage II or III TNBC undergoing neoadjuvant docetaxel and carboplatin chemotherapy on a clinical trial (NCT02124902) followed by surgery were included in this study. Blood samples were prospectively collected prior to, during, and after completion of neoadjuvant chemotherapy, post-surgery, and throughout pre-specified surveillance time points (every 6 months for 5 years). Plasma samples were analyzed using Guardant Reveal powered by Infinity, a tissue-free, multiomic assay that interrogates differentially methylated regions of DNA using an applied bioinformatics filter optimized to detect breast cancer DNA from non-cancerous cell-free DNA and estimate quantitative tumor fraction. Samples were sequenced and resulted blinded to the patients’ clinical data. Results: A total of 120 patients with evaluable plasma samples were included in this study. At baseline prior to chemotherapy, ctDNA was detected in 75% (54/72) of patients. Baseline ctDNA detection was higher in node-positive cancers (100%; 29/29) versus node-negative cancers (58%; 25/43; p<0.0001). ctDNA quantity was higher in node-positive cancers (p<0.0001), and in stage III versus stage II disease (p=0.0179). During surveillance follow-up, the sensitivity for detecting any distant metastatic recurrence was 71% (5/7). Among patients with samples drawn within one year of recurrence, the sensitivity for detecting distant recurrence was 83% (5/6) with a sample-level specificity of 98% (186/190) and a patient-level specificity of 94% (63/67). ctDNA detection prior to recurrence had a median lead time of 5.0 months (range, 4.6-5.6). Notably, the detection of ctDNA during follow-up was associated with worse recurrence-free (HR 23.3, 95% CI 2.70-201, p<0.0001) and distant-recurrence free survival (HR 29.3, 95% CI 1.86-461, p<0.0001). Conclusions: ctDNA is detected with a tissue-free MRD assay with high sensitivity and specificity for distant recurrence in early-stage TNBC. Furthermore, the presence of ctDNA during follow-up is a prognostic indicator. Additional prospective studies are needed to assess the clinical utility of ctDNA monitoring in early-stage TNBC. Citation Format: Foluso O. Ademuyiwa, Cynthia X. Ma, Katherine Weilbaecher, Rama Suresh, Lindsay Peterson, Ron Bose, Nusayba Bagegni, Caron E. Rigden, Ashley Frith, Katherine Clifton, Andrew Davis, Derek Dustin, Mingyang Cai. Evaluating circulating tumor DNA (ctDNA) as a prognostic biomarker utilizing a tissue-free epigenomic assay in early-stage triple negative breast cancer (TNBC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2420.

Abstract B059: Epigenetically defined sub-clonal heterogeneity drives therapy resistance in triple-negative breast cancer

Abstract Background: Triple-negative breast cancer (TNBC) is a highly aggressive and heterogeneous disease with an average survival of less than 50%. Although immunotherapy is showing promising results, many patients do not respond, and chemotherapy remains the main treatment. High TNBC recurrence rates are in part due to its molecular heterogeneity. Here, we sought to characterise the sub-clonal and spatial diversity of chemotherapy resistant TNBC to identify the epigenetic hallmarks of lethal sub-populations of cells that may induce cell state transition, therapy resistance and ultimately metastatic progression. Methods: Sixteen primary untreated triple-negative breast tumours (eight chemotherapy sensitive (ChemoS) and eight chemotherapy resistant (ChemoR, who died of their disease within three years)) were subjected to cell-matched high-depth multiome single nuclei RNA- and ATAC-sequencing. Four tumours were subject to spatial RNA and ATAC profiling. Fourteen TNBC PDX models generated from patients with residual disease post NAC were subjected to single nuclei sequencing and used for validation. Meta-clustering of epithelial cells from the RNA profiles was used to identify sub-populations of cells harbouring distinct epigenetic and transcriptomic features enriched in ChemoR patients. Findings were validated in the BrighTNess TNBC NAC clinical trial and in chemotherapy treated TNBC patients from the SCAN-B study. Results: Unsupervised meta-clustering (MC) of epithelial cells in G1 phase of the cell cycle identified nine MC’s across the dataset, including a distinct immune-like MC. Two MCs were dominated by cells from ChemoR patients and were defined by hypoxia, EMT, and extra-cellular matrix signalling. Sub-populations of cells in the ChemoR dominant MCs were underpinned by both promoter and distal differential chromatin accessibility, that mapped to H3K27ac enhancer sites, suggesting a role of enhancer regulation of MC transcription. Spatial ATAC and RNA profiling of a subset of tumours identified spatially epigenetically defined regions that showed distinct chromatin profiles, suggesting that the epigenetic regulation of genes that define the MCs are spatially distinct. ChemoR clusters were independently reproduced in a cohort of fourteen residual disease TNBC PDXs. Genes pertaining to the immune-like MC were associated with a good prognosis in independent cohorts of contemporary treated TNBC (SCAN-B) and with response to NAC in the BrighTNess trial. Marker genes from ChemoR dominant MCs were associated with poor prognosis and significant levels of residual disease following NAC. Conclusions: Our multi-modal integrative analysis reveals unprecedented insight into the role of epigenetically defined spatially distinct, sub-clonal populations that pre-exist in treatment naïve TNBC and predict therapy response. This suggests pre-existing populations of cells have already acquired the epigenetic footprints and hence transcriptomic features to allow them to evade therapy. Citation Format: Fatemeh Ahmadimoghari, Yu Zhang, Ioanna Mavrommatis, Shafali Thakur, Chris Starling, Ioannis Roxanis, Syed Haider, Rachael Natrajan. Epigenetically defined sub-clonal heterogeneity drives therapy resistance in triple-negative breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr B059.

The Use of Schisandrin B to Combat Triple-Negative Breast Cancers by Inhibiting NLRP3-Induced Interleukin-1β Production.

Triple-negative breast cancer (TNBC) is the most aggressive and fatal breast cancer subtype. Nowadays, chemotherapy remains the standard treatment of TNBC, and immunotherapy has emerged as an important alternative. However, the high rate of TNBC recurrence suggests that new treatment is desperately needed. Schisandrin B (Sch B) has recently revealed its anti-tumor effects in cancers such as cholangiocarcinoma, hepatoma, glioma, and multi-drug-resistant breast cancer. However, there is still a need to investigate using Sch B in TNBC treatment. Interleukin (IL)-1β, an inflammatory cytokine that can be expressed and produced by the cancer cell itself, has been suggested to promote BC proliferation and progression. In the current study, we present evidence that Sch B can significantly suppress the growth, migration, and invasion of TNBC cell lines and patient-derived TNBC cells. Through inhibition of inflammasome activation, Sch B inhibits interleukin (IL)-1β production of TNBC cells, hindering its progression. This was confirmed using an NLRP3 inhibitor, OLT1177, which revealed a similar beneficial effect in combating TNBC progression. Sch B treatment also inhibits IL-1β-induced EMT expression of TNBC cells, which may contribute to the anti-tumor response.

Association of androgen receptor and tumour-infiltrating lymphocytes with bone recurrence in triple-negative breast cancer

BackgroundAs compared to endocrine responsive breast cancer bone is less frequent site of distant recurrence in triple-negative breast cancer (TNBC). A biomarker which predicts bone recurrence would allow a more personalized treatment approach with adjuvant bisphosphonates in TNBC. Here we hypothesised that tumour expression of androgen receptor (AR) is associated with bone recurrence in TNBC. Materials and methodsPatients with operable TNBC who were treated at the Institute of Oncology Ljubljana between 2005 and 2015 and developed distant recurrence were included into our study. Nuclear expression of AR in the tissue of primary tumours was determined immunohistochemically by using the Androgen Receptor (SP107) Rabbit Monoclonal Antibody. We applied a logistic regression model to test the association between expression of AR and development of bone metastases. The model was adjusted for selected known prognostic factors and possible confounders in TNBC, including the level of the stromal tumour-infiltrating lymphocytes (sTILs). ResultsAt recurrence 45 (45 %) out of 100 patients presented with bone metastases. Additionally, seven (7 %) developed bone metastases metachronously. AR was expressed in primary tumours of 35 (35 %) women and 19 (54.3 %) developed bone recurrence. In 25 (25 %) patients sTILs were absent. Neither the proportion of AR positive cancer cells (OR = 1.00; 95 % CI 0.96–1.03; p = 1.00) nor the intensity of AR positive reaction (OR = 0.71; 95 % CI 0.02–21.4; p = 1.00) were significantly associated with bone recurrence. However, women with at least mild level of the sTILs were at significantly lower risk for bone recurrence as compared to those without any sTILs (OR = 0.01; 95 % CI < 0.01–0.08; p = 0.01). ConclusionsExpression of AR is not significantly associated with the development of bone metastases in TNBC. However, patients with absent sTILs in their primary tumours are highly susceptible for recurrence in the bone and might particularly benefit from adjuvant bisphosphonates.

Dual-Modality Imaging-Guided Manganese-Based Nanotransformer for Enhanced Gas-Photothermal Therapy Combined Immunotherapeutic Strategy Against Triple-Negative Breast Cancer.

Activating the stimulator of the interferon gene (STING) is a promising immunotherapeutic strategy for converting "cold" tumor microenvironment into "hot" one to achieve better immunotherapy for malignant tumors. Herein, a manganese-based nanotransformer is presented, consisting of manganese carbonyl and cyanine dye, for MRI/NIR-II dual-modality imaging-guided multifunctional carbon monoxide (CO) gas treatment and photothermal therapy, along with triggering cGAS-STING immune pathway against triple-negative breast cancer. This nanosystem is able to transfer its amorphous morphology into a crystallographic-like formation in response to the tumor microenvironment, achieved by breaking metal-carbon bonds and forming coordination bonds, which enhances the sensitivity of magnetic resonance imaging. Moreover, the generated CO and photothermal effect under irradiation of this nanotransformer induce immunogenic death of tumor cells and release damage-associated molecular patterns. Simultaneously, the Mn acts as an immunoactivator, potentially stimulating the cGAS-STING pathway to augment adaptive immunity, resulting in promoting the secretion of type I interferon, the proliferation of cytotoxic T lymphocytes and M2-macrophages repolarization. This nanosystem-based gas-photothermal treatment and immunoactivating therapy synergistic effect exhibit excellent antitumor efficacy both in vitro and in vivo, reducing the risk of triple-negative breast cancer recurrence and metastasis; thus, this strategy presents great potential as multifunctional immunotherapeutic agents for cancer treatment.

Roles of lncRNA in the diagnosis and prognosis of triple-negative breast cancer.

Breast cancer is a malignant tumor that seriously endangers women's lives. The prognosis of breast cancer patients differs among molecular types. Compared with other subtypes, triple-negative breast cancer (TNBC) has been a research hotspot in recent years because of its high degree of malignancy, strong invasiveness, rapid progression, easy of recurrence, distant metastasis, poor prognosis, and high mortality. Many studies have found that long non-coding RNA (lncRNA) plays an important role in the occurrence, proliferation, migration, recurrence, chemotherapy resistance, and other characteristics of TNBC. Some lncRNAs are expected to become biomarkers in the diagnosis and prognosis of TNBC, and even new targets for its treatment. Based on a PubMed literature search, this review summarizes the progress in research on lncRNAs in TNBC and discusses their roles in TNBC diagnosis, prognosis, and chemotherapy with the hope of providing help for future research.

The Combination of Radiation with PARP Inhibition Enhances Senescence and Sensitivity to the Senolytic, Navitoclax, in Triple Negative Breast Tumor Cells.

Despite significant advances in the treatment of triple-negative breast cancer, this disease continues to pose a clinical challenge, with many patients ultimately suffering from relapse. Tumor cells that recover after entering into a state of senescence after chemotherapy or radiation have been shown to develop a more aggressive phenotype, and to contribute to disease recurrence. By combining the PARP inhibitor (PARPi), talazoparib, with radiation, senescence was enhanced in 4T1 and MDA-MB-231 triple-negative breast cancer cell lines (based on SA-β-gal upregulation, increased expression of CDKN1A and the senescence-associated secretory phenotype (SASP) marker, IL6). Subsequent treatment of the radiation- and talazoparib-induced senescent 4T1 and MDA-MB231 cells with navitoclax (ABT-263) resulted in significant apoptotic cell death. In immunocompetent tumor-bearing mice, navitoclax exerted a modest growth inhibitory effect when used alone, but dramatically interfered with the recovery of 4T1-derived tumors induced into senescence with ionizing radiation and talazoparib. These findings support the potential utility of a senolytic strategy in combination with the radiotherapy/PARPi combination to mitigate the risk of disease recurrence in triple-negative breast cancer.

A serum LncRNA signature for predicting prognosis of triple-negative breast cancer

BackgroundBreast cancer is the leading causes of cancer-associated mortality among women, and triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer. Long non-coding RNAs (LncRNAs) have recently been studied to predict the prognosis of various cancers, but whether it is an effective marker in TNBC is inconclusive. MethodsWe used RNA-sequencing analysis to identify differentially expressed exosomal LncRNAs, and qRT-PCR assay was performed to verify dysregulated LncRNAs in multicenter validation cohorts. A signature, which was composed of LINC00989, CEA, and CA153, was then utilized to predict the progression and recurrence of TNBC. Kaplan-Meier analysis was applied to evaluate the prognostic values of the signature. ResultsOn the basis of RNA-sequencing analysis, we found that serum exosomal LncRNA LINC00989 was significantly up-regulated in metastatic patients of TNBC. Then LINC00989, together with clinic marker CEA and CA125, were selected to construct a prognostic signature. In both training and validation cohort, higher levels of this signature were significantly related with shorter overall and progression-free survival time. Univariate and multivariate analysis shown that the signature was the independent prognosis factor of TNBC patients. ConclusionsOur results suggested that this prognostic signature might potentially predict prognosis and recurrence of TNBC, and was worth validation in future clinical trials.

Xianlinglianxiafang Inhibited the growth and metastasis of triple-negative breast cancer via activating PPARγ/AMPK signaling pathway

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by high invasion and metastasis rates. Xian-Ling-Lian-Xia formula (XLLX) is a traditional Chinese medicine prescription widely used in China for treating TNBC. Clinical studies have shown that XLLX significantly reduces the recurrence and metastasis rate of TNBC and improves disease-free survival. However, the potential molecular mechanisms of XLLX on TNBC are not clear yet. Here, we investigated the effects of XLLX on TNBC using a mouse model and tumor cell lines. The results showed that XLLX significantly inhibited the proliferation, migration, and invasion abilities of TNBC cell lines MDA-MB-231 and 4T1 in vitro, induced apoptosis, and regulated the expression of proliferation, apoptosis, and EMT marker proteins in tumor cells. In in vivo experiments, XLLX treatment significantly reduced the progression of TNBC tumors and lung metastasis. Transcriptomics reveals that XLLX treatment significantly enriched differentially expressed genes in the peroxisome proliferator-activated receptor gamma (PPARγ) and AMP-dependent protein kinase (AMPK) signaling pathways. The western blot results confirmed that XLLX significantly upregulated the protein expression of PPARγ and p-AMPK in TNBC cells, tumors, and lung tissues. It is noteworthy that GW9662 (a PPARγ inhibitor) and Compound C (an AMPK inhibitor) partially reversed the anti-proliferation and anti-metastasis effects of XLLX in TNBC cells. Therefore, XLLX may effectively inhibit the growth and metastasis of TNBC by activating the PPARγ/AMPK signaling pathway.

A novel eight-gene Immune Cell-Associated Predictive Gene model to predict recurrence in triple-negative breast cancer.

Tumour tissue contains not only tumour cells but also some stromal cells and immune cells. This is one composition of the immune microenvironment of the tumour and causes a significant effect on the prognostic factors and recurrence of malignant tumor. In this research, single-cell RNA data from triple-negative breast cancers (TNBCs) were comprehensively analyzed and 1,527 marker genes expressed in immune cells were identified. Subsequently, RNA sequencing and clinical data from 360 patients in the Triple Negative Breast Cancer database at the Fudan University Shanghai Cancer Center (FUSCC) were divided into two groups in a 1:1 ratio, the training group and the validation group. An eight-gene Immune Cell-Associated Predictive Gene (ICAPG) model for predicting breast cancer (BC) recurrence was developed using mRNA data from the training group combined with immune cell marker genes. Based on this model, subjects were divided into two different risk level groups. The predictive power of the model was fully validated using the validation group and The Cancer Genome Atlas (TCGA) database. The localization and expression of these eight genes were then confirmed in a single-cell database. ssGSEA and CIBERSORT algorithms were used to characterize the differences in immune cell infiltration between the two different risk groups. The eight-gene ICAPG model was proven to be effective in the validation group. The low-risk group patients presented higher criterion of infiltration of CD8+ T cells and higher levels of tumour-infiltrating lymphocytes (TILs). In addition, the relationship between predictive models and homologous recombination deficiency (HRD) was explored and it was revealed that subjects from the high-risk group tended to have higher HRD values. This research established a new predictive model on the basis of immune cell marker genes that might effectively predict relapse in TNBC patients.

CTSL, a prognostic marker of breast cancer, that promotes proliferation, migration, and invasion in cells in triple-negative breast cancer.

In the world, the incidence of breast cancer has surpassed that of lung cancer, and it has become the first malignant tumor among women. Triple-negative breast cancer (TNBC) shows an extremely heterogeneous malignancy toward high recurrence, metastasis, and mortality, but there is a lack of effective targeted therapy. It is urgent to develop novel molecular targets in the occurrence and therapeutics for TNBC, and novel therapeutic strategies to block the recurrence and metastasis of TNBC. In this study, CTSL (cathepsin L) expression in tissues and adjacent tissues of TNBC patients was monitored by immunohistochemistry and western blots. The correlations between CTSL expressions and clinicopathological characteristics in the patient tissues for TNBC were analyzed. Cell proliferation, migration, and invasion assay were also performed when over-expressed or knocked-down CTSL. We found that the level of CTSL in TNBC is significantly higher than that in the matched adjacent tissues, and associated with differentiated degree, TNM Stage, tumor size, and lymph node metastatic status in TNBC patients. The high level of CTSL was correlated with a short RFS (p<0.001), OS (p<0.001), DMFS (p<0.001), PPS (p= 0.0025) in breast cancer from online databases; while in breast cancer with lymph node-positive, high level of CTSL was correlated with a short DMFS (p<0.001) and RFS (p<0.001). Moreover, in vitro experiments showed that CTSL overexpression promotes the abilities for proliferation, migration, and invasion in MCF-7 and MDA-MB-231 cell lines, while knocking-down CTSL decreases its characteristics in MDA-MB-231 cell lines. CTSL might involve into the regulation of the proliferation, invasion, and metastasis of TNBC. Thus, CTSL would be a novel, potential therapeutic, and prognostic target of TNBC.

BAG3’s Role in Indicating the Recurrence of TNBC

Triple-negative breast cancer (TNBC) cells tend to resist chemotherapy drugs, making it essential to find other therapiesto treat this aggressive cancer. The inhibition of BAG3 with YM-1 is an option because BAG3 is overexpressed in over50% of TNBC patients. BAG3 positively regulates the EGFR pathway and cell proliferation, which means inhibiting itcould reduce TNBC tumor size and lessen the recurrence of TNBC. This paper looks at xenografting BT-549 cell linesin mice and treating them with docetaxel, PBS, and YM-1 at various concentrations. Changes in the tumor are trackedusing the tumor volume and weight of the mice. The results of this work will provide a better understanding of howBAG3 inhibition impacts cell proliferation and whether it can be an indicator of TNBC recurrence. Studying YM-1gives patients more options and the chance to find the best treatment.

Recurrent versus non-recurrent triple negative breast cancer: An institutional study.

e13094 Background: Triple negative breast cancer(TNBC) is associated with(w/) worse outcomes than hormone positive cancers(CAs). We compared patients(pts) w/ TNBC recurrence at 5 years (recurrence cohort-R) against pts without(w/o)(non-recurrence cohort-N). Methods: Retrospective review. Inclusion criteria: female, 18-85, stage 1-3 TNBC, between 2010-2015. Results: The population had 219 pts. R: 36 pts(16.4%); N: 183 pts(83.6%). Most pts (95.9%) were Caucasian; Black pts: 3.2%, Asian pts: 0.5%, unknown race: 0.5%. Most pts (94.1%) were non-Hispanic; Hispanic:5.5%; unknown ethnicity:0.5%. No differences in race and ethnicity existed between (btwn) R and N. Mean age was 57.3 (standard deviation(σ) 12.8); mean BMI: 31.0 (σ 7.4); mean Charlson Comorbidity Index: 2.3 (σ 2.0); mean ECOG score: 0.3 (σ 0.6). No differences existed btwn R and N regarding the above variables. A personal hx of CA was present in 17.3% (38/219); there were no differences btwn R (19.4%) and N (16.9%), p 0.717. Genetic testing was performed in 37.8% (83/219); positive results were similar btwn R (20.0%) and N (25.0%); p 0.777. Mean tumor size was 25.2mm (σ 23.3); R had a larger mean size (34.4mm, σ 30.3) than N (23.3mm, σ 21.2), p 0.009. The most common disease stage was stage 2 (97/219, 44.3%). R had a greater proportion of stage 3 (30.6% vs 11.5%) and 2 disease (47.2% vs 43.7%) than N. There were no differences in the prevalence of histological grades (p 0.612) or subtypes (p 0.221) btwn R and N. All pts had surgery; clear margins occurred in 86.3%(189/219); margins were similar btwn R and N. Radiotherapy (RDT) was performed in 75.3% (165/219); more N pts (78.7%) received RDT than R (58.3%), p 0.01. Chemotherapy (CMT) was administered in 79.9%(175/219), and there were no differences btwn the prevalence of CMT btwn N (79.2%) and R (83.3%), p 0.575. Adjuvant CMT was received by 60.7% (133/219); there were no differences btwn N (62.3%) and R (52.8%), p 0.285. Neoadjuvant CMT was administered to 20.5% (45/219); there was a greater prevalence in R (33.3%) than in N(18.0%); p 0.038. Doxorubicin, cyclophosphamide and taxane(ACT) regimen was administered in 57.1% (125/219), while the combination of a taxane and cyclophosphamide (TC) was administered in 20.1% (44/219); there were no differences btwn R and N regarding ACT (p 0.073) or TC (p 0.069). Within 5 years of diagnosis, 31.5% were deceased. In N, 21.3% were deceased, while 83.3% were deceased in R(p&lt;0.001). The mean time-to-mortality (TTM) from CA diagnosis was 51.1 months (σ 38.9). TTM was significantly shorter in R (37.4 months, σ 20.0) than in N (61.7 months, σ 46.2), p 0.005. Conclusions: R had more advanced (stage II and III) disease, while the N had less advanced (Stage 0 and I) disease. More N pts received RDT compared w/ R pts. Neoadjuvant CMT was more prevalent in R than N. R pts had significantly larger tumor sizes than N pts. Mortality was significantly greater in the R than N, and the TTM was almost twice as long in the N than R.

Postsurgical wound management and prevention of triple-negative breast cancer recurrence with a pryoptosis-inducing, photopolymerizable hydrogel.

Surgical removal remains the predominant treatment strategy for triple-negative breast cancer (TNBC). However, risks that include high locoregional recurrence and remote metastasis threaten patient survival and quality of life after surgery. In this study, a hydrogel based on poly (ethylene glycol) dimethacrylate and sericin methacryloyl was fabricated by photopolymerization to fill the resection cavity and prevent recurrence. The obtained hydrogel exhibited mechanical properties compatible with breast tissue and facilitated postsurgical wound management by promoting tissue regeneration. The DNA methylation inhibitor decitabine (DEC) and poly (lactic-co-glycolic acid)-encapsulated phytochemical gambogic acid (GA) were loaded into the hydrogel. The as-prepared hydrogel promoted fast release of DEC and sustained release of GA, leading to gasdermin E-mediated tumor cell pyroptosis and activating antitumor immune responses. Inducing postsurgical tumor cell pyroptosis inhibited local tumor recurrence and lung metastasis. While the dual-drug-loaded hydrogel system cured less than half of tumor-bearing mice, the cured mice survived for over half a year. These findings indicate that our hydrogel system is an excellent biocompatible platform for postsurgical TNBC therapy.