Lung Metastatic Breast Cancer Research Articles

Extracellular Matrix-Trapped Bioinspired Lipoprotein Prolongs Tumor Retention to Potentiate Antitumor Immunity.

The immunomodulatory effects of many therapeutic agents are significantly challenged by their insufficient delivery efficiency and short retention time in tumors. Regarding the distinctively upregulated fibronectin (FN1) and tenascin C (TNC) in tumor stroma, herein a protease-activated FN1 and/or TNC binding peptide (FTF) is designed and an extracellular matrix (ECM)-trapped bioinspired lipoprotein (BL) (FTF-BL-CP) is proposed that can be preferentially captured by the TNC and/or FN1 for tumor retention, and then be responsively dissociated from the matrix to potentiate the antitumor immunity. The FTF-BL-CP treatment produces a 6.96-, 9.24-, 6.72-, 7.32-, and 6.73-fold increase of CD3+CD8+ T cells and their interferon-γ-, granzyme B-, perforin-, and Ki67-expressing subtypes versus the negative control, thereby profoundly eliciting the antitumor immunity. In orthotopic and lung metastatic breast cancer models, FTF-BL-CP produces notable therapeutic benefits of retarding tumor growth, extending survivals, and inhibiting lung metastasis. Therefore, this ECM-trapping strategy provides an encouraging possibility of prolonging tumor retention to potentiate the antitumor immunity for anticancer immunotherapy.

Tenascin-C Activation of Lung Fibroblasts in a 3D Synthetic Lung Extracellular Matrix Mimic.

The lung extracellular matrix (ECM) maintains the structural integrity of the tissue and regulates the phenotype and functions of resident fibroblasts. Lung-metastatic breast cancer alters these cell-ECM interactions, promoting fibroblast activation. There is a need for bio-instructive ECM models that match the ECM composition and biomechanics of the lung to study these cell-matrix interactions in vitro. Here, a synthetic, bioactive hydrogel is synthesized that mimics the native lung modulus and includes a representative distribution of the most abundant ECM peptide motifs responsible for integrin-binding and matrix metalloproteinase (MMP)-mediated degradation in the lung, which enables quiescent culture of human lung fibroblasts (HLFs). Stimulation with transforming growth factor β1 (TGF-β1), metastatic breast cancer conditioned media (CM), or tenascin-C-derived integrin-binding peptide activated hydrogel-encapsulated HLFs demonstrates multiple environmental methods to activate HLFs in a lung ECM-mimicking hydrogel. This lung hydrogel platform is a tunable, synthetic approach to studying the independent and combinatorial effects of ECM in regulating fibroblast quiescence and activation.

Sequential targeting dual-responsive magnetic nanoparticle for improved therapy of lung metastatic breast cancer

Lung metastatic breast cancer is a leading cause of cancer-related death in women and difficult to treat due to non-specific drug delivery. Herein a sequential targeting dual-responsive magnetic nanoparticle was fabricated, where Fe3O4 nanoparticle was used as magnetic core, then sequentially coated with tetraethyl orthosilicate, bis[3-(triethoxy-silyl)propyl] tetrasulfide, and 3-(trimethoxysilyl) propylmethacrylate to afford -C = C- on the surface for further polymerisation with acrylic acid, acryloyl-6-ethylenediamine-6-deoxy-β-cyclodextrin using N, N-bisacryloylcy- stamine as cross-linker, obtaining pH/redox dual-responsive magnetic nanoparticle (MNPs-CD) to delivery doxorubicin (DOX) for suppressing lung metastatic breast cancer. Our results suggested DOX-loaded nanoparticle could target the lung metastases site by sequential targeting, in which they first be delivered to the lung and even the metastatic nodules through size-driven, electrical interaction, and magnetic field-guided mechanisms, then be effectively internalised into the cancer cells followed by intelligently triggering DOX release. MTT analysis demonstrated DOX-loaded nanoparticle exhibited high anti-tumour activity against 4T1 and A549 cells. 4T1 tumour-bearing mice were employed to confirm the higher specific accumulation in lung and improved anti-metastatic therapy efficiency of DOX by focussing an extracorporeal magnetic field on the biological target. Our findings suggested the as-proposed dual-responsive magnetic nanoparticle offered a prerequisite to inhibit lung metastasis of breast cancer tumours.

"Cluster Bomb" Based Bismuth Nano-in-Micro Spheres Formed Dry Powder Inhalation for Thermo-Radio Sensitization Effects of Lung Metastatic Breast Cancer.

Lung metastatic breast cancer (LMBC) is mainly diagnosed through CT imaging and radiotherapy could be the most common method in the clinic to inhibit tumor proliferation. While the sensitivity of radiotherapy is always limited due to the hypoxic tumor microenvironment and high doses of irradiation easily induce systemic cytotoxicity. Metal-based materials applied as radiosensitizers have been widely investigated to improve efficiency and reduce the doses of irradiation. Herein, it is aimed to overcome these problems by designing biodegradable lipid-camouflaged bismuth-based nanoflowers (DP-BNFs) as both a photo-thermo-radiosensitizer to develop a novel photothermal therapy (PTT) and radiotherapy combination strategy for LMBC treatment. To achieve effective lung deposition, "Cluster Bomb" structure-based DP-BNFs nano-in-micro dry powder inhalation (DP-BNF@Lat-MPs) are formulated through spray-dried technology. The DP-BNFs "cluster" in the microsphere to improve their tumor-targeted lung deposition with a high fine particle fraction followed by burst releasing of DP-BNFs for targeting delivery and LMBC therapy. The DP-BNF@Lat-MPs exhibit excellent photothermal conversion efficiency, radiotherapy enhancement, and CT imaging ability in vitro, which synergistically inhibit cell proliferation and metastasis. In vitro and in vivo data prove that combining PTT and radiotherapy with DP-BNF@Lat-MPs as a thermo-radio dual-sensitizer significantly enhances LMBC tumor metastasis inhibition with good biocompatibility and low toxicity.

Lipid-coated bismuth nanoflower as the thermos-radio sensiti for therapy of lung metastatic breast cancer: Preparation, optimisation, and characterisation.

Lung metastatic breast cancer (LMBC) leads to a large number of deaths in women with breast cancer, and radiotherapy has been considered the common assay for tumour therapy except for surgery. However, radiotherapy still faces problems of low efficiency due to resistance and easily induced side effects. Here, the authors designed lipid-decorated bismuth-based nanoflowers (DP-BNFs) as both a radiosensitiser and a photothermal therapy agent for LMBC treatment. The BNFs were prepared by oxidation of bismuth nitrate and subsequent reduction using sodium borohydride. The preparation parameters and formulation of DP-BNFs were optimised via a single-factor experiment, with the factors including reaction temperature, a molar ratio of reducing agents, and the types and amount of decorated lipid materials. The result indicated that the BNFs prepared at 170°C with the Bi/NaBH4 ratio of 1:0.7 exhibited the best yield and particle size around 160nm. After being spray dried with lactose to prepare dry powder inhalation (DP-BNF@Lat-MPs), their effects on improving therapeutic efficiency of the radiotherapy and photothermal therapy combination were measured using the western blot assay to determine the tumour apoptosis. In a word, DP-BNF@Lat-MPs could be a novel inhalable integrated microsphere that provides a new possibility for thermoradiotherapy of LMBC.

Clinical outcomes and a nomogram for de novo metastatic breast cancer with lung metastasis: a population-based study

To better understand the clinical characteristics of newly diagnosed lung metastatic breast cancer (LMBC) and quantify its prognosis, we retrieved data on patients with LMBC from the Surveillance, Epidemiology, and End Results database. Eligible patients were randomly assigned to training and validation cohorts (ratio 7:3) to establish a nomogram using the Cox proportional hazards regression model. In total, 4310 patients with LMBC were enrolled, including 52.4% (2259/4310) HR+/HER2−, 17.6% (757/4310) HR+/HER2+, 10.8% (467/4310) HR−/HER2+, and 19.2% (827/4310) HR−/HER2− subtype patients. Inclinations of lung and brain involvement in HR−/HER2+ and HR−/HER2− subgroups, liver involvement in the HER2 overexpressing subgroup, and bone involvement in the HR-positive subgroup were detected in the LMBC population. Regarding prognosis, HR+/HER2+ subtype patients presented the most favorable profile (mOS 35.0 months, 95% CI 30.1–39.9), while HR−/HER2− patients exhibited the worst (mOS 11.0 months, 95% CI, 10.0–11.9). A nomogram was developed in the training cohort and validated internally (C-index 0.70) and externally (C-index 0.71), suggestive of decent performance. This study assessed the clinical outcomes associated with molecular subtypes, metastatic patterns, and surgical intervention and provided a robust nomogram for the estimation of survival probabilities, which are promising for the management of LMBC in clinical practice.

C3a elicits unique migratory responses in immature low-density neutrophils.

Neutrophils represent the immune system's first line of defense and are rapidly recruited into inflamed tissue. In cancer associated inflammation, phenotypic heterogeneity has been ascribed to this cell type, whereby neutrophils can manifest anti- or pro-metastatic functions depending on the cellular/micro-environmental context. Here, we demonstrate that pro-metastatic immature low-density neutrophils (iLDNs) more efficiently accumulate in the livers of mice bearing metastatic lesions compared with anti-metastatic mature high-density neutrophils (HDNs). Transcriptomic analyses reveal enrichment of a migration signature in iLDNs relative to HDNs. We find that conditioned media derived from liver-metastatic breast cancer cells, but not lung-metastatic variants, specifically induces chemotaxis of iLDNs and not HDNs. Chemotactic responses are due to increased surface expression of C3aR in iLDNs relative to HDNs. In addition, we detect elevated secretion of cancer-cell derived C3a from liver-metastatic versus lung-metastatic breast cancer cells. Perturbation of C3a/C3aR signaling axis with either a small molecule inhibitor, SB290157, or reducing the levels of secreted C3a from liver-metastatic breast cancer cells by short hairpin RNAs, can abrogate the chemotactic response of iLDNs both in vitro and in vivo, respectively. Together, these data reveal novel mechanisms through which iLDNs prefentially accumulate in liver tissue harboring metastases in response to tumor-derived C3a secreted from the liver-aggressive 4T1 breast cancer cells.

Prognostic factors and survival outcomes according to tumor subtype in patients with breast cancer lung metastases.

BackgroundReports on the incidence and prognoses of lung metastases when diagnosing breast cancer patients with different subtypes are limited. Our study investigated the effect of molecular sub-typing stratification on the prognoses of lung metastatic breast caner patients.MethodsPatients with breast cancer and lung metastases were identified from Surveillance, Epidemiology and End Results population-based data between 2010 and 2015. Univariate and multivariate Cox regression analyses were performed to identify risk factors and prognoses, overall survival (OS) and breast cancer-specific survival for patients with breast cancer lung metastases.ResultsWe identified 6,516 patients with lung metastatic breast cancer, representing 1.7% of the entire cohort and 30.4% of the subset with metastatic disease. This included 2,940 hormone receptor (HR)+/HER2− patients, 852 HR+/HER2+ patients, 547 HR−/HER2+ patients and 983 triple-negative patients. The median OS for all lung metastatic patients was 13 months. Multivariate analysis revealed that those lung metastatic breast cancer patients of older age (>80), black race, with poorly differentiated tumors, carcinoma histology, triple-negative subtype, more metastatic sites and no surgery, and no chemotherapy showed significantly poor survival, both overall and breast cancer-specific.ConclusionsOur findings show that molecular sub-type and more metastatic sites might have significant influence on the incidence and prognosis of breast cancer lung metastases. We also identified several prognostic factors that could guide therapy selection in the treatment of lung metastatic patients.

Co-delivery of TRAIL and siHSP70 using hierarchically modular assembly formulations achieves enhanced TRAIL-resistant cancer therapy

The combined therapy of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and heat shock protein 70-targeting siRNA (siHSP70) has shown an improved anti-tumor effect on TRAIL-resistant tumor. However, vehicles to co-deliver these two biopharmaceuticals are challenging because of the distinct location of their targets on the cell surface and in the cytosol. Here we developed a hierarchically modular assembly formulation (TH-s-RSC) via the copper-free click reaction to co-encapsulate the positively-charged TRAIL and negatively-charged siHSP70 and release them in the extracellular space and cytoplasm. We demonstrate that TH-s-RSC can protect the packaged biopharmaceuticals through its hyaluronic acid shell in vivo, and sequentially release TRAIL in response to extracellular molecular including hyaluronidase (HAase) and matrix metalloproteinase 2 (MMP2), followed by the release of siHSP70 triggered by the reductive conditions in the cytoplasm. We showed that the complementary activity of TRAIL and siHSP70 exhibited superior synergistic anticancer efficacy in both A549 lung cancer xenograft models and 4T1 lung metastatic breast cancer models, compared to either treatment alone. Our strategy provides a promising platform for safe and effective co-delivery and dual-site targeting of biopharmaceuticals in cancer treatment that may be applicable in the future.

PO-034 Inhibition of lung carcinoma growth and pulmonary metastasis with DIMATE as single agent and in combination with cisplatin

IntroductionAberrant cell proliferation in NSCLC causes an aberrant redox state that leads to the production of toxic reactive species and aldehydes. To keep oxidative stress until a threshold, above which oxidative damage can be detrimental to cell viability, cancer cells must actively upregulate multiple antioxidant systems. The aldehyde dehydrogenase (ALDH) gene superfamily encodes enzymes that are critical for certain life processes and detoxification of numerous endogenous and exogenous aldehyde substrates, including pharmaceuticals and environmental pollutants.Material and methodsIn this study, a meta-analysis was conducted based on multiple microarray data from The Cancer Genome Atlas (TCGA) and gene expression omnibus (GEO) repositories, spanning lung adenocarcinoma and lung squamous cell carcinoma datasets. Twenty-six NSCLC cell lines with different oncogenic driver alterations were used to analyse the molecular and cellular consequences of ALDHs inhibition. Preclinical studies with orthotopic xenografts of NSCLC and lung metastatic breast cancer were performed to evaluate the effect of the inhibition of ALDH class 1 and class3 activity on tumour growth.Results and discussionsHere, we found that increased expression of aldehyde dehydrogenase isoenzymes ALDH1A1, ALDH1A3 and ALDH3A1 in human NSCLC tumours has a strong impact on chemotherapy resistance and patient overall survival, correlating with poor prognosis. We showed that inhibition of class 1 and class 3 ALDH activity with the novel irreversible ALDH1/3 inhibitor DIMATE suppresses tumour growth in orthotopic human lung cancer xenograft model and inhibits lung metastasis of human breast cancer in athymic nude mice. Accumulation of HNE-protein adducts and depletion of intracellular GSH are main responsible of DIMATE-induced cell death. Consistently, we found that alteration of tumour redox balance further enhances sensitivity of NSCLC cells to DIMATE. Finally, we demonstrate that the combination of DIMATE with Cisplatin-induced ROS generation in an orthotopic lung cancer model can significantly enhance the cytotoxicity of these two drugs in NSCLC cells with a synergistic promotion of cell death and marginal systemic toxicity in animals.ConclusionTargeting the detoxification machinery of ALDHs constitutes a novel therapeutic avenue for NSCLC. Patients with increased expression of ALDH1 or ALDH3 might greatly benefit from a combination therapy that include drugs interfering with the activity of these enzymes to overcome patient-specific drug resistance.

76P - Serological Detection of Matrix Metalloproteinase Generated Fragments of Elastin and Type IV Collagen may be Biomarkers of Lung Metastatic Breast Cancer

Introduction: Lung is one of the main sites of metastasis in human breast cancer. The pathogenesis of lung cancer is similar to chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF) where elastin and type IV collagen play critical roles and excess tissue turnover mediated by elevated levels of matrix metalloproteinase (MMP)-9 and -12 is a pathological feature. The aim was to evaluate whether novel biochemical markers of elastin and type IV collagen degradation mediated by MMP-9 and -12 may provide information in relation to lung tissue alterations during pulmonary diseases.

Expression of Immunomodulatory Neutrophil-activating Protein of Helicobacter pylori Enhances the Antitumor Activity of Oncolytic Measles Virus

Helicobacter pylori neutrophil-activating protein (NAP) is a major virulence factor and powerful inducer of inflammatory reaction and Th1-polarized immune response. Here, we evaluated the therapeutic efficacy of measles virus (MV) strains engineered to express secretory NAP forms against metastatic breast cancer. Recombinant viruses encoding secretory NAP forms (MV-lambda-NAP and MV-s-NAP) efficiently infect and destroy breast cancer cells by cell-to-cell viral spread and large syncytia formation independently of hormone receptor status. Intrapleural administration of MV-s-NAP doubled the median survival in a pleural effusion xenograft model: 65 days as compared to 29 days in the control group (P < 0.0001). This therapeutic effect correlated with a brisk Th1 type cytokine response in vivo. Secretory NAP was expressed at high levels by infected tumor cells and increased tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-12/23 cytokine concentrations were detected in the pleural effusion. In an aggressive model of lung metastatic breast cancer, MV-lambda-NAP and MV-s-NAP also significantly improved survival of the treated animals (P < 0.05) as compared to the control MV strain. These data suggest that potent immunomodulators of bacterial origin, such as H. pylori NAP, can enhance the antitumor effect of oncolytic viruses and support the feasibility and potential of a combined viroimmunotherapy approach.

Peroxiredoxin 2 specifically regulates the oxidative and metabolic stress response of human metastatic breast cancer cells in lungs

Little is known about metastatic pathways that are specific to the lung rather than other organs. We previously showed that antioxidant proteins such as peroxiredoxins were specifically upregulated in lung metastatic breast cancer cells. We hypothesize that cancer cells that live under aerobic conditions, as might be the case in lungs, protect themselves against the damage caused by reactive oxygen species (ROS). To examine this hypothesis, we studied the role of peroxiredoxin-2 (PRDX2) in lung vs bone metastasis formation. A metastatic variant of MDA-MB-435 breast cancer cells that specifically metastasize to lungs (435-L3) was transduced with short hairpin RNAs to specifically silence PRDX2. Conversely, a bone metastatic variant of MDA-MB-231 cells (BO2) was stably transfected to overexpress PRDX2. The 435-L3 cells silenced for PRDX2 were significantly more sensitive to H(2)O(2)-induced oxidative stress than the parental and scrambled transfected cells. BO2/PRDX2 cells produced less ROS than BO2/green fluorescent protein control cells under oxidative stress. Moreover, PRDX2 knockdown inhibited the growth of 435-L3 cells in the lungs, whereas lymph node metastasis remained unaffected. In contrast, PRDX2 overexpression in bone metastatic BO2 breast cancer cells led to drastic inhibition of the skeletal tumor burden and reduction of bone destruction. Furthermore, PRDX2 expression in breast cancer cells was associated with a glucose-dependent phenotype, different from bone metastatic cells. Overall, our results strongly suggest that PRDX2 is a targetable 'metabolic adaptor' driver protein implicated in the selective growth of metastatic cells in the lungs by protecting them against oxidative stress.

Abstract P2-18-01: Treatment of Human Breast Tumors by Oncolytic Herpes Simplex Virus

Abstract Background: Breast cancer is the most common cancer for women in the world, although current therapies have improve the outcome, the breast cancer deaths are still the second highest among cancers in women. Metastasis to the lung frequently results in severe complications, and currently, there are no curative treatment options available for patients with metastetic breast cancer. Oncolytic viral therapy is a new strategy for treating cancers. Replication-competent herpes simplex virus (HSV) can infect, replicate and kill tumor cells by a direct cytopathic effect and then spread within the tumor cells. In this study, the cytotoxicity of HSV vectors, G47Δ were examined on different human breast cancer cell lines, immortalized human breast cell lines, and primary culture normal human breast cells. Pulmonary metastasis model of breast cancer was constructed and the treatment effect of G47D was evaluated. Methods: Human BC cell lines SK-BR-3, MDA-MB-453, and MCF-7, immortalized breast cells MCF-10A and 76N-tert, and cultured primary normal breast cells HMEC1 and HMEC2 were cultured in vitro and infected with G47Δ at different multiplicities of infection (MOIs). The viability of infected cells was measured. G47Δ-infected cells were identified using X-gal histochemistry for LacZ expression. A pulmonary metastatic BC model was established with Balb-c nude mice by tail vein injection of MDA-MB-435 cells. Treatment was initiated 21 days after injection of tumor cells, with the virus group treated twice weekly with four total i.v. administrations of G47Δ (2*107pfu/100ul) and on day 60 mice were sacrificed and tumor nodules on the surface of the lung counted after Indian ink staining. Results: G47Δ was highly cytotoxic to BC cells and 76N-tert in vitro at very low MOI (0.01). For all of these cell lines, more than 85% and 95% cells were killed at MOI=0.01 and MOI=0.1 respectively on the fifth day after infection. However, both HMEC1 and HMEC2 cells were viable even 5 days after infection. A slight effect was found with MCF-10A, of which only 7% and 24.5% were killed at MOI=0.01 and MOI=0.1 respectively on day 5. In the pulmonary metastatic model, the average number of surface lung tumor nodules in the G47Δ group was one, but ten in the control group. X-gal staining illustrated viral replication and spread in the tumor cells in vitro and in vivo. Conclusions: Recently constructed oncolytic HSV vector G47Δ was effective at killing human breast cancer cells and immortalized breast cells, but didn't infect the normal breast cells. The i.v. administrations of G47Δ could effectively cure metastatic breast cancer. Our studies demonstrated that as a novel therapeutic agents, G47Δ was safe and exhibited superior antitumor effects, thereby making it appropriate for human clinical trials. It is worth noting that G47Δ was effective for lung metastatic breast cancer, which could assure the long survival rate of patients with advance breast cancer. The efficiency of G47Δ warrants consideration of clinical application. (Key words) breast cancer; cancer therapy; herpes simplex virus; oncolytic virus; gene therapy; Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-18-01.

Abstract 75: Electron transport chain-dependent oxygen consumption in metastatic breast cancer cells

Abstract Although mitochondrial ATP-production deficiency has been historically implicated to account for the Warburg effect or “aerobic” glycolysis in cancer cells, recent observations indicate the presence of functionally competent mitochondria in cancer cells. We hypothesized that a lung-metastatic breast cancer cell line with intact mitochondrial membrane potential would exhibit mitochondria-dependent oxygen consumption. A murine breast cancer cell line (4T1) was used to evaluate mitochondrial membrane potential by JC-1 fluorescence microscopy. Oxygen consumption by cancer cells in suspension was determined using a dissolved oxygen meter. Fluorescence microscopy of JC-1 red aggregates indicated the presence of highly polarized mitochondria in 4T1 cells, suggesting that these cells are capable of oxidative phosphorylation. Oxygen consumption rate of 4T1 cells suspended in 10 mM glucose-containing medium equilibrated with 21% oxygen (258 µM dissolved oxygen) at 25°C was 2.98 nmol/min/million cells (Control). Oxygen consumption was significantly enhanced by carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), a mitochondrial uncoupler (7.88 nmol/min/million cells, p&amp;lt;0.05 vs. Control). Myxothiazol (a respiratory chain complex III inhibitor) inhibited the oxygen consumption (0.75 nmol/min/million cells; p&amp;lt;0.05 vs. Control), indicating the presence of a functional electron transport chain. A nitric oxide donor also inhibited oxygen consumption (0.84 nmol/min/million cells; p&amp;lt;0.05 vs. Control). These data indicate the presence of oxidative phosphorylation-competent mitochondria in breast cancer cells. We conclude that cancer cells possess respiration-capable mitochondria and their “aerobic” glycolysis may not reflect a compensatory mechanism for ATP generation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 75.

Systemic efficacy of oncolytic adenoviruses in imagable orthotopic models of hormone refractory metastatic breast cancer

Conditionally replicating oncolytic adenoviruses represent a promising developmental strategy for the treatment of cancer refractory to current treatments, such as hormone refractory metastatic breast cancer. In clinical cancer trials, adenoviral agents have been well tolerated, but gene transfer has been insufficient for clinical benefit. One of the main reasons may be the deficiency of the primary adenovirus receptor, and therefore viral capsid modifications have been employed. Another obstacle to systemic delivery is rapid clearance of virus by hepatic Kupffer cells and subsequent inadequate bioavailability. In this study, we compared several capsid-modified oncolytic adenoviruses for the treatment of breast cancer with and without Kupffer cell inactivation. Replication deficient capsid-modified viruses were analyzed for their gene transfer efficacy in vitro in breast cancer cell lines and clinical samples and in vivo in orthotopic models of breast cancer. The effect of Kupffer cell depleting agents on gene transfer efficacy in vivo was evaluated. An aggressive lung metastatic model was developed to study the effect of capsid-modified oncolytic adenoviruses on survival. Capsid-modified viruses displayed increased gene transfer and cancer cell killing in vitro and resulted in increased survival in an orthotopic model of lung metastatic breast cancer in mice. Biodistribution of viruses was favorable, tumor burden and treatment response could be monitored repeatedly. Kuppfer cell inactivation led to enhanced systemic gene delivery, but did not increase the survival of mice. These results facilitate clinical translation of oncolytic adenoviruses for the treatment of hormone refractory metastatic breast cancer.