Highlights from the 2023 International Meeting on the Molecular Biology of Hepatitis B virus.

Since its discovery in 1965, our understanding of the hepatitis B virus (HBV) replication cycle and host immune responses has increased markedly. In contrast, our knowledge of the molecular biology of hepatitis delta virus (HDV), which is associated with more severe liver disease, is less well understood. Despite the progress made, critical gaps remain in our knowledge of HBV and HDV replication and the mechanisms underlying viral persistence and evasion of host immunity. The International HBV Meeting is the leading annual scientific meeting for presenting the latest advances in HBV and HDV molecular virology, immunology, and epidemiology. In 2023, the annual scientific meeting was held in Kobe, Japan and this review summarises some of the advances presented at the Meeting and lists gaps in our knowledge that may facilitate the development of new therapies.

Tea polyphenol-derived nanomedicine for targeted photothermal thrombolysis and inflammation suppression.

Thrombotic diseases impose a significant global health burden, and conventional drug-based thrombolytic therapies are encumbered by the risk of bleeding complications. In this study, we introduce a novel drug-free nanomedicine founded on tea polyphenols nanoparticles (TPNs), which exhibits multifaceted capabilities for localized photothermal thrombolysis. TPNs were synthesized through a one-pot process under mild conditions, deriving from the monomeric epigallocatechin-3-gallate (EGCG). Within this process, indocyanine green (ICG) was effectively encapsulated, exploiting multiple intermolecular interactions between EGCG and ICG. While both TPNs and ICG inherently possessed photothermal potential, their synergy significantly enhanced photothermal conversion and stability. Furthermore, the nanomedicine was functionalized with cRGD for targeted delivery to activated platelets within thrombus sites, eliciting robust thrombolysis upon laser irradiation across diverse thrombus types. Importantly, the nanomedicine's potent free radical scavenging abilities concurrently mitigated vascular inflammation, thus diminishing the risk of disease recurrence. In summary, this highly biocompatible multifunctional nanomaterial holds promise as a comprehensive approach that combines thrombolysis with anti-inflammatory actions, offering precision in thrombosis treatment.

Deep learning and big data mining for Metal-Organic frameworks with high performance for simultaneous desulfurization and carbon capture.

Carbon capture and desulfurization of flue gases are crucial for the achievement of carbon neutrality and sustainable development. In this work, the "one-step" adsorption technology with high-performance metal-organic frameworks (MOFs) was proposed to simultaneously capture the SO2 and CO2. Four machine learning algorithms were used to predict the performance indicators (NCO2+SO2, SCO2+SO2/N2, and TSN) of MOFs, with Multi-Layer Perceptron Regression (MLPR) showing better performance (R2 = 0.93). To address sparse data of MOF chemical descriptors, we introduced the Deep Factorization Machines (DeepFM) model, outperforming MLPR with a higher R2 of 0.95. Then, sensitivity analysis was employed to find that the adsorption heat and porosity were the key factors for SO2 and CO2 capture performance of MOF, while the influence of open alkali metal sites also stood out. Furthermore, we established a kinetic model to batch simulate the breakthrough curves of TOP 1000 MOFs to investigate their dynamic adsorption separation performance for SO2/CO2/N2. The TOP 20 MOFs screened by the dynamic performance highly overlap with those screened by the static performance, with 76 % containing open alkali metal sites. This integrated approach of computational screening, machine learning, and dynamic analysis significantly advances the development of efficient MOF adsorbents for flue gas treatment.

Oxalate anions-intercalated NiFe layered double hydroxide as a highly active and stable electrocatalyst for alkaline seawater oxidation.

Seawater electrolysis is gaining recognition as a promising method for hydrogen production. However, severe anode corrosion caused by the high concentration of chloride ions (Cl-) poses a challenge for the long-term oxygen evolution reaction. Herein, an anti-corrosion strategy of oxalate anions intercalation in NiFe layered double hydroxide on nickel foam (NiFe-C2O42- LDH/NF) is proposed. The intercalation of these highly negatively charged C2O42- serves to establish electrostatic repulsion and impede Cl- adsorption. In alkaline seawater, NiFe-C2O42- LDH/NF requires an overpotential of 337 mV to gain the large current density of 1000 mA cm-2 and operates continuously for 500 h. The intercalation of C2O42- is demonstrated to significantly boost the activity and stability of NiFe LDH-based materials during alkaline seawater oxidation.

CD95 promotes stemness of colorectal cancer cells by lncRNA MALAT1.

Colorectal cancer (CRC) is the second most fatal cancer. Many studies have shown that cancer stemness contributes to resistance to conventional chemotherapy and poor prognosis. However, the mechanisms involved in maintaining cancer stemness in CRC are still obscure and few clinical drugs were used to target cancer stemness. Previous studies had reported CD95 increases the stemness of cancer cells with long-term stimulation of exogenous agonist CD95 ligand (CD95L). However, the expression of CD95L is relative low in certain human tumor tissues. In this study, we found that CD95 was highly expressed in CRC cells, and in vitro it promoted the tumorsphere formation, chemotherapy resistance and in vivo tumor growth without stimulation of exogenous CD95L. Mechanistically, the bulk and single-cell RNA-sequencing results suggested that CD95 promotes stemness of CRC cells through upregulation of long non-coding RNAs metastasis-associated lung adenocarcinoma transcript 1 (lncRNA MALAT1). MALAT1 knockdown inhibited CD95-induced tumorsphere formation and chemotherapy resistance. In summary, our findings reveal that CD95 has the capability to modulate cancer stemness via the action of the lncRNA MALAT1. Targeting CD95 may be a promising strategy to inhibit cancer stemness in CRC.

A computational spatial whole-Cell model for hepatitis B viral infection and drug interactions.

Despite a vaccine, hepatitis B virus (HBV) remains a world-wide source of infections and deaths. We develop a whole-cell computational platform combining spatial and kinetic models describing the infection cycle of HBV in a hepatocyte host. We simulate key parts of the infection cycle with this whole-cell platform for 10 min of biological time, to predict infection progression, map out virus-host and virus-drug interactions. We find that starting from an established infection, decreasing the copy number of the viral envelope proteins shifts the dominant infection pathway from capsid secretion to re-importing the capsids into the nucleus, resulting in more nuclear-localized viral covalently closed circular DNA (cccDNA) and boosting transcription. This scenario can mimic the consequence of drugs designed to manipulate viral gene expression. Mutating capsid proteins facilitates capsid destabilization and disassembly at nuclear pore complexes, resulting in an increase in cccDNA copy number. However, excessive destabilization leads to premature cytoplasmic disassembly and does not increase the cccDNA counts. Finally, our simulations can predict the best drug dosage and its administration timing to reduce the cccDNA counts. Our adaptable computational platform can be parameterized to study other viruses and identify the most central viral pathways that can be targeted by drugs.

CpG-Conjugated Silver Nanoparticles as a Multifunctional Nanomedicine to Promote Macrophage Efferocytosis and Repolarization for Atherosclerosis Therapy.

Atherosclerosis (AS) is a major contributor to cardiovascular diseases, necessitating the development of novel therapeutic strategies to alleviate plaque burden. Macrophage efferocytosis, the process by which macrophages clear apoptotic and foam cells, plays a crucial role in plaque regression. However, this process is impaired in AS lesions due to the overexpression of CD47, which produces a "do not eat me" signal. In this study, we investigated the potential of CpG, a toll-like receptor 9 agonist, to enhance macrophage efferocytosis for AS therapy. We demonstrated that CpG treatment promoted the engulfment of CD47-positive apoptotic cells and foam cells by macrophages. Mechanistically, CpG induced a metabolic shift in macrophages characterized by enhanced fatty acid oxidation and de novo lipid biosynthesis, contributing to its pro-efferocytic effect. To enable in vivo application, we conjugated CpG on silver nanoparticles (AgNPs) to form CpG-AgNPs, which could protect CpG from biological degradation, promote its cellular uptake, and release CpG in response to intracellular glutathione. Combining the intrinsic antioxidative and anti-inflammatory abilities of AgNPs, such nanomedicine displayed multifunctionalities to simultaneously promote macrophage efferocytosis and repolarization. In an ApoE-/- mouse model, intravenous administration of CpG-AgNPs effectively targeted atherosclerotic plaques and exhibited potent therapeutic efficacy with excellent biocompatibility. Our study provides valuable insights into CpG-induced macrophage efferocytosis and highlights the potential of CpG-AgNPs as a promising therapeutic strategy for AS.

Exploration of prognosis and immunometabolism landscapes in ER+ breast cancer based on a novel lipid metabolism-related signature.

Introduction: Lipid metabolic reprogramming is gaining attention as a hallmark of cancers. Recent mounting evidence indicates that the malignant behavior of breast cancer (BC) is closely related to lipid metabolism. Here, we focus on the estrogen receptor-positive (ER+) subtype, the most common subgroup of BC, to explore immunometabolism landscapes and prognostic significance according to lipid metabolism-related genes (LMRGs). Methods: Samples from The Cancer Genome Atlas (TCGA) database were used as training cohort, and samples from the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC), Gene Expression Omnibus (GEO) datasets and our cohort were applied for external validation. The survival-related LMRG molecular pattern and signature were constructed by unsupervised consensus clustering and least absolute shrinkage and selection operator (LASSO) analysis. A lipid metabolism-related clinicopathologic nomogram was established. Gene enrichment and pathway analysis were performed to explore the underlying mechanism. Immune landscapes, immunotherapy and chemotherapy response were further explored. Moreover, the relationship between gene expression and clinicopathological features was assessed by immunohistochemistry. Results: Two LMRG molecular patterns were identified and associated with distinct prognoses and immune cell infiltration. Next, a prognostic signature based on nine survival-related LMRGs was established and validated. The signature was confirmed to be an independent prognostic factor and an optimal nomogram incorporating age and T stage (AUC of 5-year overall survival: 0.778). Pathway enrichment analysis revealed differences in immune activities, lipid biosynthesis and drug metabolism by comparing groups with low- and high-risk scores. Further exploration verified different immune microenvironment profiles, immune checkpoint expression, and sensitivity to immunotherapy and chemotherapy between the two groups. Finally, arachidonate 15-lipoxygenase (ALOX15) was selected as the most prominent differentially expressed gene between the two groups. Its expression was positively related to larger tumor size, more advanced tumor stage and vascular invasion in our cohort (n = 149). Discussion: This is the first lipid metabolism-based signature with value for prognosis prediction and immunotherapy or chemotherapy guidance for ER+ BC.

Hepatitis B Virus Capsid: The Core in Productive Entry and Covalently Closed Circular DNA Formation.

Hepatitis B virus (HBV) relies on the core protein (HBc) to establish productive infection, as defined by the formation of the covalently closed circularized DNA (cccDNA), as well as to carry out almost every step of the lifecycle following cccDNA formation. Multiple copies of HBc form an icosahedral capsid shell that encapsidates the viral pregenomic RNA (pgRNA) and facilitates the reverse transcription of pgRNA to a relaxed circular DNA (rcDNA) within the capsid. During infection, the complete HBV virion, which contains an outer envelope layer in addition to the internal nucleocapsid containing rcDNA, enters human hepatocytes via endocytosis and traffics through the endosomal compartments and the cytosol to deliver its rcDNA to the nucleus to produce cccDNA. In addition, progeny rcDNA, newly formed in cytoplasmic nucleocapsids, is also delivered to the nucleus in the same cell to form more cccDNA in a process called intracellular cccDNA amplification or recycling. Here, we focus on recent evidence demonstrating differential effects of HBc in affecting cccDNA formation during de novo infection vs. recycling, obtained using HBc mutations and small molecule inhibitors. These results implicate a critical role of HBc in determining HBV trafficking during infection, as well as in nucleocapsid disassembly (uncoating) to release rcDNA, events essential for cccDNA formation. HBc likely functions in these processes via interactions with host factors, which contributes critically to HBV host tropism. A better understanding of the roles of HBc in HBV entry, cccDNA formation, and host species tropism should accelerate ongoing efforts to target HBc and cccDNA for the development of an HBV cure and facilitate the establishment of convenient animal models for both basic research and drug development.

Graph Neural Tree: A novel and interpretable deep learning-based framework for accurate molecular property predictions.

Determining various properties of molecules is a critical step in drug discovery. Recently, with the improvement of large heterogeneous datasets and the development of deep learning approaches, more and more scientists have turned their attention to neural network-based virtual preliminary screening to reduce the time and monetary cost of drug discovery. However, the poor interpretability of deep learning masks causality, so models' conclusions are often beyond the comprehension of human users, which reduces the credibility of the model and makes it difficult for chemists to further narrow the huge chemical space based on models' results. Thus, this study develops a novel framework consisting of Graph Neural Networks for feature extraction, Curriculum-Based Learning Strategies for optimization, and a Learning Binary Neural Tree (LBNT) for prediction, to improve the performance of neural networks and reveal their decision-making process to chemists. The framework encodes molecular graph data with graph neural networks (GNNs), then retrains the encoder with curriculum-based learning strategies to reduce uncertainty and improve accuracy, and finally uses LBNT as the predictor, which joint retrains with the encoder after independently training, for prediction and visualization. The framework is validated on the public datasets and compared to single GNNs with normal training strategies as well as GNN encoders with common machine learning predictors instead of the LBNT predictor. The result reveals that the proposed framework enhances the point prediction accuracy of the completely trained GNN and reduces its uncertainty through curriculum-based learning, and further improves the accuracy by combining LBNT. Besides, compared with common machine learning tools, the LBNT predictor generally has the best performance because of joint retraining with the GNN encoder. The decision-making process of LBNT is also better and easier to explain than that of other models.

A general strategy for protein affinity-ligand oriented-immobilization and screening for bioactive compounds.

Natural products containing complex mixtures of potentially bioactive compounds are a major source of new drugs, however, conventional screening for active compounds is a time-consuming and inefficient process. Here, we reported that a facile and efficient protein affinity-ligand oriented-immobilization strategy based on the SpyTag/SpyCatcher(ST/SC) chemistry, was used for bioactive compound screening. Two ST-fused model proteins, that is, GFP (green fluorescent protein) and PqsA (a critical enzyme in the quorum sensing pathway of Pseudomonas aeruginosa), were used to verify the feasibility of this screening method. GFP, as the capturing protein model, was ST-labeled and anchored at a specific orientation onto the surface of activated agarose coupled with SC protein via ST/SC self-ligation. The affinity carriers were characterized by infrared spectroscopy and fluorography. The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses. Although the alkaline stability of the affinity carriers was not ideal, its pH stability was acceptable under pH < 9. The general preparation strategy of this affinity carriers was validated by replacing GFP with PqsA, and PqsA inhibitor, 2-amino-6-fluorobenzoic acid, was successfully isolated from the fermentation broth. The proposed strategy can immobilize protein ligands in one-step and screen compounds that interact specifically with the ligands.

M2 tumor-associated macrophage mediates the maintenance of stemness to promote cisplatin resistance by secreting TGF-ß1 in esophageal squamous cell carcinoma.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a deadly gastrointestinal malignancy, and chemotherapy resistance is a key factor leading to its poor prognosis. M2 tumor-associated macrophages (M2-TAMs) may be an important cause of chemoresistance in ESCC, but its exact mechanism is still unclear. METHODS: In order to study the role of M2-TAMs in ESCC chemoresistance, CCK-8, clone formation assay, flow cytometric apoptosis assay, qRT-PCR, western blotting, and serum-free sphere formation assays were used. In vivo animal experiments and human ESCC tissues were used to confirm the findings. RESULTS: In vitro and in vivo animal experiments, M2-TAMs reduced the sensitivity of ESCC cells to cisplatin. Mechanistically, M2-TAMs highly secreted TGF-ß1 which activated the TGFßR1-smad2/3 pathway to promote and maintain the stemness characteristic of ESCC cells, which could inhibit the sensitivity to cisplatin. Using TGFß signaling inhibitor SB431542 or knockdown of TGFßR1 could reverse the cisplatin resistance of ESCC cells. In 92 cases of human ESCC tissues, individuals with a high density of M2-TAMs had considerably higher levels of TGF-ß1. These patients also had worse prognoses and richer stemness markers. CONCLUSION: TGF-ß1 secreted from M2-TAMs promoted and maintained the stemness characteristic to induce cisplatin resistance in ESCC by activating the TGFß1-Smad2/3 pathway.

Characterization of Intracellular Precore-Derived Proteins and Their Functions in Hepatitis B Virus-Infected Human Hepatocytes.

Hepatitis B virus (HBV) precore protein is not essential for viral replication but is thought to facilitate chronic infection. In addition to the secreted precore products, including the hepatitis B e antigen (HBeAg) and PreC protein, intracellular precore-derived proteins in HBV-infected human hepatocytes remain poorly characterized, and their roles, if any, remain largely unknown. Here, we detected multiple precore derivatives, including the nonprocessed precursor p25 and the processing intermediate p22, in HBV-infected human hepatocytes as well as human hepatoma cells overexpressing the HBV precore protein. Both p25 and p22 showed phosphorylated and unphosphorylated forms, which were located in different intracellular compartments. Interestingly, precore expression was associated with decreases in intracellular HBV core protein (HBc) and secreted DNA-containing virions but was also associated with an increase in secreted empty virions. The decrease in HBc by precore could be attributed to cytosolic p22, which caused HBc degradation, at least in part by the proteasome, and consequently decreased HBV pregenomic RNA packaging and DNA synthesis. In addition, cytosolic p22 formed chimeric capsids with HBc in the cell, which were further secreted in virions. In contrast, the PreC antigen, like HBeAg, was secreted via the endoplasmic reticulum (ER)-Golgi secretory pathway and was thus unable to form capsids in the cell or be secreted in virions. Furthermore, p25, as well as p22, were secreted in virions from HBV-infected human hepatocytes and were detected in the sera of HBV-infected chimpanzees. In summary, we have detected multiple intracellular precore-derived proteins in HBV-infected human hepatocytes and revealed novel precore functions in the viral life cycle. IMPORTANCE Chronic hepatitis B remains a worldwide public health issue. The hepatitis B virus (HBV) precore protein is not essential for HBV replication but may facilitate viral persistence. In this study, we have detected multiple precore protein species in HBV-infected human hepatocytes and studied their functions in the HBV life cycle. We found that the HBV precore proteins decreased intracellular HBV core protein and reduced secretion of complete virions but enhanced secretion of empty virions. Interestingly, the cytosolic precore protein species formed chimeric capsids with the core protein and were secreted in virions. Our results shed new light on the functions of intracellular precore protein species in the HBV life cycle and have implications for the roles of precore proteins in HBV persistence and pathogenesis.

HPV 16 E6 promotes growth and metastasis of esophageal squamous cell carcinoma cells in vitro.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is one of the most lethal malignancies worldwide. Increasing evidence suggests that human papillomavirus (HPV) infection may be associated with the etiology of ESCC. However, the precise role of HPV in ESCC remains unclear. METHODS AND RESULTS: Proliferation and apoptosis of ESCC cells upon infection with HPV16 E6 were detected using CCK-8 assays and Western blot analyses. The migration rate was measured with a wound healing assay, and a Transwell Matrigel invasion assay was used to detect the invasive ability. RT-qPCR was performed to detect the expression of E6AP, p53, and miR-34a. The proliferation rates were significantly higher in HPV16E6-transfected cell groups compared with the negative control groups. Bax protein expression was downregulated in HPV16E6-treated groups compared to the controls. The wound healing and Transwell Matrigel invasion assays indicated that HPV16 E6 infection could increase ESCC cell migration and invasion. Furthermore, E6AP, p53 and miR-34a expression were decreased in HPV16 E6-transfected cell lines. CONCLUSION: Our results not only provide evidence that HPV16 E6 promotes cell proliferation, migration, and invasion in ESCC, but also suggests a correlation between HPV infection and E6AP, p53 and miR-34a expression. Consequently, HPV16 E6 may play an important role in ESCC development.

Replication-driven HBV cccDNA loss in chimeric mice with humanized livers.

Hepatitis B virus (HBV) infection is largely noncytopathic and requires the establishment of covalently closed circular DNA (cccDNA), which is considered stable in the nuclei of infected cells. Although challenging, approaches to directly target cccDNA molecules or kill infected cells are recommended to eliminate cccDNA. Herein, cccDNA levels were investigated in HBV-infected chimeric mice with humanized livers. HBV-infected cells support robust replication, progressively retain viral products, and head for cytopathic destruction and cccDNA loss. It is difficult for infected cells to retain cccDNA and remain noncytopathic. Replication-driven cccDNA loss is observed at both phases of spread of and persistent infection. The cccDNA replenishment is required to compensate for cccDNA loss. Blocking cccDNA replenishment pathways reduces cccDNA levels by >100-fold. These results prove an unconventional cccDNA elimination strategy that does not directly target cccDNA but aims to transform spontaneous cccDNA loss into progressive cccDNA elimination by blocking cccDNA replenishment.

Polo-like kinase 1 as a potential therapeutic target and prognostic factor for various human malignancies: A systematic review and meta-analysis.

Objective: The overexpression of polo-like kinase 1 (PLK-1) has been found in a broad spectrum of human tumors, making it an attractive prognostic tumor biomarker. Nowadays, PLK-1 is considered a cancer therapeutic target with clinical therapeutic value. The aim of the present study was to systematically review the prognostic and therapeutic value of PLK-1 in different malignant neoplasms. Methods: A systematic literature search of the Cochrane Library, PubMed, Web of Science, and China National Knowledge Internet (CNKI) databases was conducted between December 2018 and September 2022. In total, 41 published studies were screened, comprising 5,301 patients. We calculated the pooled odds ratios (ORs) and corresponding 95%CIs for the clinical parameters of patients included in these studies, as well as the pooled hazard ratios (HRs) and corresponding 95% CIs for 5-year overall survival (OS). Results: Our analysis included 41 eligible studies, representing a total of 5,301 patients. The results showed that overexpression of PLK-1 was significantly associated with poor OS (HR, 1.57; 95% CI, 1.18-2.08) and inferior 5-year disease-free survival/relapse-free survival ((HR, 1.89; 95% CI, 1.47-2.44). The pooled analysis showed that PLK-1 overexpression was significantly associated with lymph node metastasis, histological grade, clinical stages (p < 0.001 respectively), and tumor grade (p < 0.001). In digestive system neoplasms, PLK-1 overexpression was significantly associated with histopathological classification, primary tumor grade, histological grade, and clinical stages (p = 0.002, p = 0.001, p < 0.0001, respectively). In breast cancer, PLK-1 was significantly associated with 5-year overall survival, histological grade, and lymph node metastasis (p < 0.001, p = 0.003, p < 0.001, respectively). In the female reproductive system, PLK-1 was significantly associated with clinical stage (p = 0.011). In the respiratory system, PLK-1 was significantly associated with clinical stage (p = 0.021). Conclusion: Our analysis indicates that high PLK-1 expression is associated with aggressiveness and poor prognosis in malignant neoplasms. Therefore, PLK-1 may be a clinically valuable target for cancer treatment.

A novel Bayesian ensembling model for wind power forecasting.

Precise and robust wind power prediction can effectively alleviate the problem caused by the randomness and volatility of wind power. Ensemble learning can successfully improve forecasting precision and robustness, and quantify the uncertainty of the prediction. This paper presents a new ensemble probabilistic forecasting framework, based on modified randomized maximum a posteriori (MAP) sampling technique, echo state network (ESN) and generalized mixture (GM) function to bring superior forecasting results. The proposed model first trains a set of independent ESN models for probabilistic forecasting using the modified randomized MAP sampling technique, and then dynamically weighs and ensembles the base model forecasting through the GM function. The proposed model and other benchmark models have been implemented on four wind power datasets from different places to illustrate the advantage of the proposed method. The compared result indicates that the suggested model outperforms some state-of-the-art models and can successfully achieve dynamic ensemble probabilistic prediction.

Crosstalk between the peripheral nervous system and breast cancer influences tumor progression.

Recent studies have shown that peripheral nerves play an important role in the progression of breast cancer. Breast cancer cells (BCCs) promote local peripheral nerve growth and branching by secreting neuroactive molecules, including neurotrophins and axon guidance molecules (AGMs). Sympathetic nerves promote breast cancer progression, while parasympathetic and sensory nerves mainly have anti-tumor effects in the progression of breast cancer. Specifically, peripheral nerves can influence the progression of breast cancer by secreting neurotransmitters not only directly binding to the corresponding receptors of BCCs, but also indirectly acting on immune cells to modulate anti-tumor immunity. In this review, we summarize the crosstalk between breast cancer and peripheral nerves and the roles of important neuroactive molecules in the progression of breast cancer. In addition, we summarize indicators, including nerve fiber density and perineural invasion (PNI), that may help determine the prognosis of breast cancer based on current research results, as well as potential therapeutic approaches, such as ß-blockers and retroviral-mediated genetic neuroengineering techniques, that may enhance the prognosis of breast cancer. In addition, we propose suggestions for future research priorities based on a current lack of knowledge in this area.