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1.
J Drug Target ; 29(3): 294-299, 2021 03.
Article in English | MEDLINE | ID: mdl-32886538

ABSTRACT

Much of the bacterial anticancer therapy being developed relies on the ability of bacteria to specifically colonise tumours. Initial attempts to translate promising Salmonella enterica Typhimurium (S. Typhimurium) preclinical results to the clinical setting failed, primarily due to lack of tumour colonisation and the significant toxicities from systemically administered Gram-negative bacteria. To address the difference in results between preclinical experiments performed in mice with transplant tumours and clinical trials in human volunteers with autochthonous tumours, a genetically engineered mouse model of breast cancer (BALB-neuT) was utilised to develop a strain of virulence-attenuated S. Typhimurium capable of robust colonisation of autochthonous tumours. Several genes that code for bacterial surface molecules, responsible for signalling a toxic immune response against the bacteria, were mutated. The resulting S. Typhimurium strain, BCT2, allowed non-toxic intravenous administration of 3 × 106 colony forming units of bacteria in tumour-burdened mice when combined with a vascular disruption agent to induce intratumoral necrotic space and facilitate bacterial colonisation.


Subject(s)
Genetic Engineering , Mammary Neoplasms, Experimental/microbiology , Salmonella typhimurium/physiology , Animals , Female , Mammary Neoplasms, Experimental/therapy , Mice , Mice, Inbred BALB C , Mice, Transgenic , Salmonella typhimurium/genetics , Salmonella typhimurium/pathogenicity
2.
Biomater Sci ; 8(13): 3712-3719, 2020 Jul 07.
Article in English | MEDLINE | ID: mdl-32495762

ABSTRACT

Attenuated bacteria-mediated tumor targeting diagnosis will provide a novel strategy for further cancer treatments owing to the intrinsic facultative anaerobic characteristic of bacteria and rapid proliferation in the tumor sites. In this work, we firstly investigate the in vivo behaviour of the attenuated Salmonella typhimurium (S. typhimuriumΔppGpp/lux) after intravenous injection. S. typhimurium exhibits rapid proliferation in tumor sites after three days of injection through bioluminescence imaging, the Luria-Bertani plate and the Gram-staining assay. In contrast, S. typhimurium does not proliferate in the normal tissues and could be excreted from the body of mice. Afterwards, a targeting peptide ubiquicidin (UBI) labeled with fluorescent dye Cy5.5 or radionuclide 125I was intravenously injected into the mice with or without S. typhimurium treatments for in vivo fluorescence imaging and single-photon emission computed tomography (SPECT/CT) imaging, respectively. The results show that the peptide UBI could specifically target the two independent bacteria-infected tumor models, the 4T1 murine breast cancer model and the CT26 mouse colon cancer model, realizing the sensitive multimodal imaging of tumors. Such a strategy (bacteria-mediated tumor targeting) may further improve the sensitivity to early diagnosis of tumors. We hope that our developed strategy could further be extended to cancer theranostics in the future, bringing good news for cancer patients.


Subject(s)
Biocompatible Materials/chemistry , Breast Neoplasms/diagnostic imaging , Optical Imaging , Peptides/chemistry , Ribosomal Proteins/chemistry , Salmonella typhimurium/isolation & purification , Animals , Biocompatible Materials/administration & dosage , Breast Neoplasms/microbiology , Carbocyanines/chemical synthesis , Carbocyanines/chemistry , Cell Proliferation , Female , Iodine Radioisotopes , Mammary Neoplasms, Experimental/diagnostic imaging , Mammary Neoplasms, Experimental/microbiology , Mice , Molecular Structure , Peptides/administration & dosage , Ribosomal Proteins/administration & dosage
3.
Biochem Biophys Res Commun ; 514(4): 1147-1153, 2019 07 05.
Article in English | MEDLINE | ID: mdl-31103266

ABSTRACT

The hypoxic region microenvironment reduces the susceptibility of the cancer cells to radiotherapy and anticancer drugs of the solid tumors. However, the reduced oxygen surroundings provide an appreciable habitat for anaerobic bacteria to colonize and proliferate. Herein, we present a biocompatible bacteriabased system that can deliver poly(lactic-co-glycolic acid)(PLGA) nanoparticles(PLGA NPs) specifically targeting into solid tumor to achieve precision imaging and treatment. In our strategy, anaerobic bacterium Bifidobacterium longum (B. longum) that colonizes selectively in hypoxic regions of animal body was successfully used as a vehicle to conjugate with PLGA NPs and transported into solid tumors. To improve the efficacy and specificity of tumor therapy, low-boiling point perfluorohexane (PFH) liquid was wrapped in the core of PLGA NPs (PFH/PLGA NPs), which could increase the deposition of energy by affecting the acoustic environment of the tumor and destroy cells after liquid-gas phase transition during High Intensity Focused Ultrasound (HIFU) irradiation. This strategy shows an effective diagnosis and treatment integration for giving stronger imaging, longer retention period and more effective tumor therapy.


Subject(s)
Antineoplastic Agents/pharmacology , Bifidobacterium longum/chemistry , Fluorocarbons/chemistry , Nanoparticles/chemistry , Optical Imaging , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Animals , Female , Fluorocarbons/administration & dosage , Humans , Mammary Neoplasms, Experimental/diagnostic imaging , Mammary Neoplasms, Experimental/microbiology , Mammary Neoplasms, Experimental/therapy , Mice , Mice, Inbred BALB C , Mice, Nude , Nanoparticles/administration & dosage , Polylactic Acid-Polyglycolic Acid Copolymer/administration & dosage , Tumor Cells, Cultured
4.
Oncotarget ; 7(25): 37513-37523, 2016 Jun 21.
Article in English | MEDLINE | ID: mdl-27175584

ABSTRACT

Salmonella is a Gram-negative, facultative anaerobe that is a common cause of host intestinal infections. Salmonella grows under aerobic and anaerobic conditions, and it has been proven capable of inhibiting tumor growth. However, the molecular mechanism by which Salmonella inhibits tumor growth is still unclear. Angiogenesis plays an important role in the development and progression of tumors. We investigated the antitumor effect of Salmonella in a syngeneic murine tumor model. Hypoxia-inducible factor-1 (HIF-1)α plays a significant role in tumor angiogenesis. We examined the molecular mechanism by which Salmonella regulated vascular endothelial growth factor (VEGF), which is an important angiogenic factor. The expression of VEGF in tumor cells was decreased by treatment with Salmonella. The conditioned medium from Salmonella-treated cells inhibited the proliferation of endothelial cells. Salmonella inhibited the expression of HIF-1α as well as downregulated its upstream signal mediator protein kinase B (AKT). Salmonella significantly inhibited tumor growth in vivo, and immunohistochemical studies of the tumors revealed decreased intratumoral microvessel density. These results suggest that Salmonella therapy, which exerts anti-angiogenic activities, represents a promising strategy for the treatment of tumors.


Subject(s)
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Mammary Neoplasms, Experimental/blood supply , Mammary Neoplasms, Experimental/microbiology , Melanoma, Experimental/blood supply , Melanoma, Experimental/microbiology , Salmonella enterica/physiology , Vascular Endothelial Growth Factor A/metabolism , Animals , Culture Media, Conditioned , Down-Regulation , Endothelial Cells/cytology , Endothelial Cells/metabolism , Endothelial Cells/microbiology , Female , Humans , Mammary Neoplasms, Experimental/metabolism , Melanoma, Experimental/metabolism , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Neovascularization, Pathologic/metabolism , Neovascularization, Pathologic/microbiology , Neovascularization, Pathologic/pathology , Salmonella enterica/metabolism , Signal Transduction , Transfection
5.
Oncotarget ; 7(23): 35169-80, 2016 Jun 07.
Article in English | MEDLINE | ID: mdl-27145267

ABSTRACT

Therapeutic attenuated strains of Salmonella Typhimurium target and eradicate tumors in mouse models. However, the mechanism of S. Typhimurium for tumor targeting is still poorly understood. We performed a high-throughput screening of single-gene deletion mutants of S. Typhimurium in an orthotopic, syngeneic murine mammary model of breast cancer. The mutants under selection in this system were classified into functional categories to identify bacterial processes involved in Salmonella accumulation within tumors. Niche-specific genes involved in preferential tumor colonization were identified and exemplars were confirmed by competitive infection assays. Our results show that the chemotaxis gene cheY and the motility genes motAB confer an advantage for colonization of Salmonella within orthotopic syngeneic breast tumors. In addition, eutC, a gene belonging to the ethanolamine metabolic pathway, also confers an advantage for Salmonella within tumors, perhaps by exploiting either ethanolamine or an alternative nutrient in the inflamed tumor environment.


Subject(s)
Genes, Bacterial/genetics , Mammary Neoplasms, Experimental/microbiology , Salmonella Infections, Animal/genetics , Salmonella typhimurium/growth & development , Salmonella typhimurium/genetics , Animals , Female , High-Throughput Nucleotide Sequencing , Mice , Mice, Inbred BALB C
6.
Anticancer Res ; 35(2): 843-9, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25667465

ABSTRACT

BACKGROUND/AIM: Cancer treatment with attenuated Salmonella enterica Typhimurium (S. Typhimurium) has gained momentum in recent years. However, the effectiveness of this treatment has not been explored in autochthonous models. We report the efficacy of S. Typhimurium in mice with autochthonous mammary tumors. MATERIALS AND METHODS: S. Typhimurium attenuated by deletion of cyclic adenosine monophosphate signaling, SalpNG.1, was injected into female BALB-neuT tumor-bearing mice. Mice were monitored for efficacy and sacrificed for mechanistic studies. RESULTS: In treated mice, seven-week post-treatment tumor burden was reduced by 85% and median survival was increased by 88%. Efficacy was correlated with increased tumor-infiltrating CD8 and natural killer cells. In addition, SalpNG.1 treatment caused a systemic increase of monocytic myeloid-derived suppressor cells that accumulated to high numbers within tumor tissue. Bacteria were not detected in tumor tissue, suggesting that the observed efficacy was due to a systemic rather than a tumor-specific effect of the bacteria. CONCLUSION: S. Typhimurium treatment reduces tumor burden and increases survival in an autochthonous breast cancer model.


Subject(s)
Mammary Neoplasms, Experimental/pathology , Salmonella typhimurium/physiology , Tumor Burden , Animals , Female , Mammary Neoplasms, Experimental/microbiology , Mice , Mice, Inbred BALB C
7.
Lett Appl Microbiol ; 59(6): 580-6, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25163827

ABSTRACT

UNLABELLED: Clostridium sporogenes ATCC 3584 is an obligate anaerobe that has been reported to possess excellent tumour-targeting capacity. Here, we use Cl. sporogenes as a vector to deliver IL-12, a potent antitumour cytokine that bears numerous antitumour properties but that has limited clinical applications due to its strong toxicity when delivered systemically. In this study, Cl. sporogenes was genetically engineered to secrete murine IL-12, and its antitumour efficacy and toxicity were investigated in a murine EMT6 mammary carcinoma model. After intravenous injection, Cl. sporogenes was able to selectively settle and reproduce in the tumours without encroaching on normal tissues, resulting in a clear delay of tumour growth and a 14·3% cure rate. Importantly, the mice showed no obvious toxicity-associated side effects, such as diarrhoea and weight loss, during the treatment process. The significant antitumour efficacy and low toxicity of this treatment may be explained by the selective tumour-targeting properties of Cl. sporogenes and by the sustained release of IL-12 accompanying bacterial proliferation. This moderate local IL-12 concentration would not induce the severe response in the entire body, that is inevitable when IL-12 is administered directly. SIGNIFICANCE AND IMPACT OF THE STUDY: Interleukin-12 (IL-12) is a potent antitumour cytokine, but it is toxic when administrated systemically. This study demonstrates that murine IL-12 can be systemically delivered to hypoxic sites in solid tumours by Clostridium sporogenes, producing a clear delay in tumour growth and a 14·3% cure rate in a mouse tumour model. Importantly, there is no obvious toxicity associated with IL-12 during the treatment process. This result may be accounted for by the excellent tumour-targeting capacity of Cl. sporogenes, targeting IL-12 directly to the tumour site instead of to the entire body.


Subject(s)
Clostridium/genetics , Interleukin-12/metabolism , Mammary Neoplasms, Experimental/microbiology , Mammary Neoplasms, Experimental/therapy , Animals , Cell Hypoxia , Clostridium/growth & development , Clostridium/metabolism , Female , Genetic Therapy , Genetic Vectors , Interferon-gamma/biosynthesis , Interferon-gamma/blood , Interleukin-12/genetics , Mammary Neoplasms, Experimental/immunology , Mice, Inbred BALB C , Recombinant Proteins/metabolism
8.
J Biol Chem ; 289(19): 13054-65, 2014 May 09.
Article in English | MEDLINE | ID: mdl-24668817

ABSTRACT

The intracellular metabolism and cytostatic activity of the anticancer drug gemcitabine (2',2'-difluoro-2'-deoxycytidine; dFdC) was severely compromised in Mycoplasma hyorhinis-infected tumor cell cultures. Pronounced deamination of dFdC to its less cytostatic metabolite 2',2'-difluoro-2'-deoxyuridine was observed, both in cell extracts and spent culture medium (i.e. tumor cell-free but mycoplasma-containing) of mycoplasma-infected tumor cells. This indicates that the decreased antiproliferative activity of dFdC in such cells is attributed to a mycoplasma cytidine deaminase causing rapid drug catabolism. Indeed, the cytostatic activity of gemcitabine could be restored by the co-administration of tetrahydrouridine (a potent cytidine deaminase inhibitor). Additionally, mycoplasma-derived pyrimidine nucleoside phosphorylase (PyNP) activity indirectly potentiated deamination of dFdC: the natural pyrimidine nucleosides uridine, 2'-deoxyuridine and thymidine inhibited mycoplasma-associated dFdC deamination but were efficiently catabolized (removed) by mycoplasma PyNP. The markedly lower anabolism and related cytostatic activity of dFdC in mycoplasma-infected tumor cells was therefore also (partially) restored by a specific TP/PyNP inhibitor (TPI), or by exogenous thymidine. Consequently, no effect on the cytostatic activity of dFdC was observed in tumor cell cultures infected with a PyNP-deficient Mycoplasma pneumoniae strain. Because it has been reported that some commensal mycoplasma species (including M. hyorhinis) preferentially colonize tumor tissue in cancer patients, our findings suggest that the presence of mycoplasmas in the tumor microenvironment could be a limiting factor for the anticancer efficiency of dFdC-based chemotherapy. Accordingly, a significantly decreased antitumor effect of dFdC was observed in mice bearing M. hyorhinis-infected murine mammary FM3A tumors compared with uninfected tumors.


Subject(s)
Antimetabolites, Antineoplastic , Bacterial Proteins/metabolism , Breast Neoplasms , Deoxycytidine/analogs & derivatives , Mammary Neoplasms, Experimental , Mycoplasma Infections/enzymology , Mycoplasma hyorhinis/enzymology , Pyrimidine Phosphorylases/metabolism , Animals , Antimetabolites, Antineoplastic/pharmacokinetics , Antimetabolites, Antineoplastic/pharmacology , Breast Neoplasms/drug therapy , Breast Neoplasms/metabolism , Breast Neoplasms/microbiology , Cell Line, Tumor , Deoxycytidine/pharmacokinetics , Deoxycytidine/pharmacology , Female , Humans , Mammary Neoplasms, Experimental/drug therapy , Mammary Neoplasms, Experimental/metabolism , Mammary Neoplasms, Experimental/microbiology , Mice , Tetrahydrouridine/pharmacokinetics , Tetrahydrouridine/pharmacology , Thymidine/metabolism , Tumor Microenvironment/drug effects , Gemcitabine
9.
Br J Cancer ; 108(11): 2281-90, 2013 Jun 11.
Article in English | MEDLINE | ID: mdl-23640395

ABSTRACT

BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are present in large numbers in blood of mice and humans with cancer, and they strongly inhibit T-cell and natural killer (NK) cell responses, at young and old age. We found that a highly attenuated bacterium Listeria monocytogenes (Listeria(at))-infected MDSC and altered the immune-suppressing function of MDSC. METHODS: Young (3 months) and old (18 months) BALB/cByJ mice with metastatic breast cancer (4T1 model) were immunised with Listeria(at) semi-therapeutically (once before and twice after tumour development), and analysed for growth of metastases and primary tumour, in relation to MDSC-, CD8 T-cell and NK cell responses. RESULTS: We found that Listeria(at)-infected MDSC, which delivered Listeria(at) predominantly to the microenvironment of metastases and primary tumours, where they spread from MDSC into tumour cells (infected tumour cells will ultimately become a target for Listeria-activated immune cells). Immunotherapy with Listeria(at) significantly reduced the population of MDSC in blood and primary tumours, and converted a remaining subpopulation of MDSC into an immune-stimulating phenotype producing IL-12, in correlation with significantly improved T-cell and NK cell responses to Listeria(at) at both ages. This was accompanied with a dramatic reduction in the number of metastases and tumour growth at young and old age. CONCLUSIONS: Although preclinical studies show that immunotherapy is less effective at old than at young age, our study demonstrates that Listeria(at)-based immunotherapy can be equally effective against metastatic breast cancer at both young and old age by targeting MDSC.


Subject(s)
Listeria monocytogenes/immunology , Mammary Neoplasms, Experimental/therapy , Age Factors , Animals , Female , Immunotherapy/methods , Interleukin-12/biosynthesis , Interleukin-12/immunology , Killer Cells, Natural/immunology , Killer Cells, Natural/microbiology , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/microbiology , Mice , Mice, Inbred BALB C , Myeloid Cells/immunology , T-Lymphocytes/immunology , T-Lymphocytes/microbiology
10.
Iran J Allergy Asthma Immunol ; 12(1): 81-5, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23454783

ABSTRACT

Breast cancer patients are susceptible to infections such as candidiasis. Due to the importance and the role of matrix metalloproteinases (MMP) in breast cancer progression and its correlation with tumor metastasis, we analyzed the serum level of MMPs -2, -3, -9 and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in breast cancer bearing mice in the presence of systemic Candida albicans infection. Female BALB/c mice were divided into 4 groups: group I had tumor + candidiasis; group II, tumor only; group III, candidiasis only and group IV as negative control. Tumor tissue was separated from stock breast cancer bearing mice and transplanted subcutaneously into the groups I and II mice. Two weeks after tumor transplantation, groups I and III were infected with Candida albicans by intravenous injection. One week after systemic infection, the sera of the experimental groups were prepared and analyzed with ELISA for MMP-2, -3, -9 and TIMP-1 levels. The results showed that the levels of MMP-3, MMP-9 and TIMP-1 were increased in groups I, II and III, as compared to the control group. However, the level of MMP-2 was decreased in mice infected with Candida albicans and in infected mice bearing tumor. These data suggest that candidiasis may have a positive effect on tumor progression and metastasis.


Subject(s)
Adenocarcinoma/microbiology , Candida albicans/immunology , Candidiasis/microbiology , Gene Expression Regulation/immunology , Mammary Glands, Animal/microbiology , Mammary Neoplasms, Experimental/microbiology , Adenocarcinoma/complications , Adenocarcinoma/immunology , Adenocarcinoma/pathology , Animals , Candidiasis/complications , Candidiasis/immunology , Candidiasis/pathology , Female , Male , Mammary Glands, Animal/immunology , Mammary Glands, Animal/pathology , Mammary Neoplasms, Experimental/complications , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/pathology , Matrix Metalloproteinase 2/blood , Matrix Metalloproteinase 2/immunology , Matrix Metalloproteinase 3/blood , Matrix Metalloproteinase 3/immunology , Matrix Metalloproteinase 9/blood , Matrix Metalloproteinase 9/immunology , Mice , Mice, Inbred BALB C , Tissue Inhibitor of Metalloproteinase-1/blood , Tissue Inhibitor of Metalloproteinase-1/immunology
11.
Appl Environ Microbiol ; 78(21): 7603-10, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22923405

ABSTRACT

Many studies have demonstrated that intravenously administered bacteria can target and proliferate in solid tumors and then quickly be released from other organs. Here, we employed the tumor-targeting property of Escherichia coli Nissle 1917 to inhibit mouse B16 melanoma and 4T1 breast tumors through the expression of azurin protein. For this purpose, recombinant azurin-expressing E. coli Nissle 1917 was developed. The levels of in vitro and in vivo azurin secretion in the engineered bacterium were determined by immunochemistry. Our results demonstrated that B16 melanoma and orthotopic 4T1 breast tumor growth were remarkably restrained and pulmonary metastasis was prevented in immunocompetent mice. It is worth noting that this therapeutic effect partially resulted from the antitumor activity of neutrophils and lymphocytes due to inflammatory responses caused by bacterial infections. No toxicity was observed in the animal during the experiments. This study indicates that E. coli Nissle 1917 could be a potential carrier to deliver antitumor drugs effectively for cancer therapy.


Subject(s)
Azurin/biosynthesis , Drug Delivery Systems , Escherichia coli , Mammary Neoplasms, Experimental/therapy , Melanoma, Experimental/therapy , Animals , Antineoplastic Agents/therapeutic use , Azurin/genetics , Azurin/therapeutic use , Cell Line, Tumor , Disease Models, Animal , Escherichia coli/classification , Escherichia coli/genetics , Escherichia coli/metabolism , Female , Genetic Engineering , Lung Neoplasms/prevention & control , Lung Neoplasms/secondary , Mammary Neoplasms, Experimental/metabolism , Mammary Neoplasms, Experimental/microbiology , Melanoma, Experimental/metabolism , Melanoma, Experimental/microbiology , Mice , Mice, Inbred BALB C , Plasmids
12.
Br J Cancer ; 101(10): 1683-91, 2009 Nov 17.
Article in English | MEDLINE | ID: mdl-19861961

ABSTRACT

BACKGROUND: An effective cancer therapeutic must selectively target tumours with minimal systemic toxicity. Expression of a cytotoxic protein using Salmonella typhimurium would enable spatial and temporal control of delivery because these bacteria preferentially target tumours over normal tissue. METHODS: We engineered non-pathogenic S. typhimurium to secrete murine TNF-related apoptosis-inducing ligand (TRAIL) under the control of the prokaryotic radiation-inducible RecA promoter. The response of the RecA promoter to radiation was measured using fluorometry and immunoblotting. TRAIL toxicity was determined using flow cytometry and by measuring caspase-3 activation. A syngeneic murine tumour model was used to determine bacterial accumulation and the response to expressed TRAIL. RESULTS: After irradiation, engineered S. typhimurium secreted TRAIL, which caused caspase-3-mediated apoptosis and death in 4T1 mammary carcinoma cells in culture. Systemic injection of Salmonella and induction of TRAIL expression using 2 Gy gamma-irradiation caused a significant delay in mammary tumour growth and reduced the risk of death by 76% when compared with irradiated controls. Repeated dosing with TRAIL-bearing Salmonella in conjunction with radiation improved the 30-day survival from 0 to 100%. CONCLUSION: These results show the pre-clinical utility of S. typhimurium as a TRAIL expression vector that effectively reduces tumour growth and extends host survival.


Subject(s)
Mammary Neoplasms, Experimental/drug therapy , Mammary Neoplasms, Experimental/microbiology , Salmonella typhimurium/physiology , TNF-Related Apoptosis-Inducing Ligand/administration & dosage , Animals , Apoptosis/drug effects , Caspase 3/genetics , Caspase 3/metabolism , Cell Line, Tumor , Disease Models, Animal , Female , Flow Cytometry , Gene Expression Regulation, Neoplastic , Immunoblotting , Immunohistochemistry , Mammary Neoplasms, Experimental/pathology , Mice , Mice, Inbred BALB C , Promoter Regions, Genetic , Rec A Recombinases/genetics , Salmonella typhimurium/genetics , Salmonella typhimurium/metabolism , TNF-Related Apoptosis-Inducing Ligand/genetics , TNF-Related Apoptosis-Inducing Ligand/metabolism
13.
Cell Microbiol ; 10(6): 1235-48, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18208564

ABSTRACT

The successful application of live bacteria in cancer therapy requires a more detailed understanding of bacterial interaction with the tumour microenvironment. Here, we analysed the effect of Escherichia coli K-12 colonization on the tumour microenvironment by immunohistochemistry and fluorescence microscopy in the murine 4T1 breast carcinoma model. We described the colonization of tumour-bearing mice, as well as the spatiotemporal distribution of E. coli K-12 in the 4T1 tumour tissue over a period of 14 days. The colonization resulted within 3 days in large avascular necrotic tissue, redistribution of hypoxic areas and an enhanced collagen IV deposition within the colonized tumour tissue, which changed the tumoral perfusion of systemically injected immunoglobulins. In addition, E. coli K-12 colonization led to the redistribution of tumour-associated macrophages, forming a granulation tissue around bacterial colonies, and also to an increase in TNFalpha and matrix metalloproteinase 9 expression. Colonization of 4T1 tumours by E. coli K-12 resulted in strong reduction of pulmonary metastatic events. These new insights will contribute to the general understanding of the tumour-microbe cross-talk and to the design of bacterial strains with enhanced anticancer efficiency.


Subject(s)
Carcinoma/therapy , Escherichia coli K12 , Mammary Neoplasms, Experimental/therapy , Animals , Carcinoma/metabolism , Carcinoma/microbiology , Carcinoma/pathology , Carcinoma/secondary , Cell Line, Tumor , Collagen Type IV/metabolism , Colony Count, Microbial , Escherichia coli K12/isolation & purification , Female , Humans , Immunohistochemistry , Liver/microbiology , Lung Neoplasms/pathology , Lung Neoplasms/secondary , Macrophages/pathology , Mammary Neoplasms, Experimental/metabolism , Mammary Neoplasms, Experimental/microbiology , Mammary Neoplasms, Experimental/pathology , Matrix Metalloproteinase 9/metabolism , Mice , Mice, Inbred BALB C , Mice, Nude , Microscopy, Fluorescence , Necrosis/pathology , Oxygen/metabolism , Spleen/microbiology , Tumor Necrosis Factor-alpha/metabolism
14.
Int J Med Microbiol ; 297(3): 151-62, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17448724

ABSTRACT

Systemic administration of microorganisms into tumor-bearing mice revealed preferential accumulation in tumors in comparison to clearance in organs such as spleen and liver. Here we compared the efficiency of tumor-specific colonization of pathogenic Salmonella typhimurium strains 14028 and SL1344 to the enteroinvasive Escherichia coli 4608-58 strain and to the attenuated Salmonella flexneri 2a SC602 strain, as well as to the uropathogenic E. coli CFT073, the non-pathogenic E. coli Top10, and the probiotic E. coli Nissle 1917 strain. All strains colonized and replicated in tumors efficiently each resulting in more than 1 x 10(8) colony-forming units per gram tumor tissue. Colonization of spleen and liver were significantly lower when E. coli strains were used in comparison to S. typhimurium and the non-pathogenic strains did not colonize those organs at all. Further investigation of E. coli Nissle 1917 showed that no drastic differences in colonization and amplification were seen when immunocompetent and immunocompromised animals were used, and we were able to show that E. coli Nissle 1917 replicates at the border of live and necrotic tumor tissue. We also demonstrated exogenously applied L-arabinose-dependent gene activation in colonized tumors in live mice. These findings will prepare the way for bacterium-mediated controlled protein delivery to solid tumors.


Subject(s)
Breast Neoplasms/therapy , Carcinoma, Intraductal, Noninfiltrating/therapy , Escherichia coli , Gene Expression , Mammary Neoplasms, Experimental/therapy , Animals , Arabinose/biosynthesis , Arabinose/genetics , Breast Neoplasms/microbiology , Carcinoma, Intraductal, Noninfiltrating/microbiology , Colony Count, Microbial , Escherichia coli/growth & development , Escherichia coli/metabolism , Female , Genetic Vectors/physiology , Liver/microbiology , Mammary Neoplasms, Experimental/microbiology , Mice , Mice, Inbred BALB C , Mice, Nude , Probiotics , Recombinant Proteins/biosynthesis , Salmonella typhimurium/growth & development , Shigella flexneri/growth & development , Species Specificity , Spleen/microbiology
15.
Cancer Res ; 66(15): 7395-400, 2006 Aug 01.
Article in English | MEDLINE | ID: mdl-16885333

ABSTRACT

Inflammation associated with bacterial infections is a risk factor for cancers in humans, yet its role in breast cancer remains poorly understood. We have previously shown that innate immune inflammatory response against intestinal bacteria is sufficient to induce colon cancer. Here we report that infecting Rag2-deficient C57BL/6 Apc(Min/+) mice with an intestinal bacterial pathogen, Helicobacter hepaticus, significantly promotes mammary carcinoma in females and enhances intestinal adenoma multiplicity by a tumor necrosis factor alpha (TNFalpha)-dependent mechanism. The mammary and intestinal tumor development as well as the increase in proinflammatory mediators is suppressed by adoptive transfer of interleukin 10-competent CD4+CD45RB(lo)CD25+ regulatory (T(R)) cells. Furthermore, prior exposure of donor mice to H. hepaticus significantly enhances antitumor potency of their T(R) cells. Interestingly, these microbially experienced T(R) cells suppress tumorigenesis more effectively in recipient mice irrespective of their tumor etiology. These data suggest that infections with enteric pathogens enhance T(R)-cell potency and protect against epithelial cancers later in life, potentially explaining paradoxical increases in cancer risk in developed countries having more stringent hygiene practices. The possibility that dysregulated gut microbial infections in humans may lead to cancer in anatomically distant organs, such as breast, highlights the need for novel immune-based strategies in cancer prevention and treatment.


Subject(s)
Adenocarcinoma/immunology , Adenocarcinoma/microbiology , Helicobacter Infections/complications , Helicobacter Infections/immunology , Helicobacter hepaticus/immunology , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/microbiology , Adenocarcinoma/genetics , Adenocarcinoma/prevention & control , Animals , DNA-Binding Proteins/deficiency , DNA-Binding Proteins/genetics , Female , Helicobacter Infections/genetics , Immunity, Innate/immunology , Immunoglobulins/genetics , Immunoglobulins/pharmacology , Immunotherapy, Adoptive , Interleukin-10/genetics , Interleukin-10/immunology , Interleukin-10/pharmacology , Intestinal Neoplasms/genetics , Intestinal Neoplasms/immunology , Intestinal Neoplasms/microbiology , Intestinal Neoplasms/prevention & control , Mammary Neoplasms, Experimental/genetics , Mammary Neoplasms, Experimental/prevention & control , Mice , Mice, Inbred C57BL , Receptors, Interleukin-2/immunology , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/pharmacology , T-Lymphocytes, Regulatory/immunology , Tumor Necrosis Factor-alpha/immunology
16.
Vopr Onkol ; 48(2): 223-6, 2002.
Article in Russian | MEDLINE | ID: mdl-12227073

ABSTRACT

Our experiments were concerned with formation of antitumor cross-resistance by Trichinella spiralis infection in rats. Prophylactic infection was followed by an insignificant decrease in the number of 3,4-benzpyrene-induced malignant fibrous histiocytoma. It also inhibited mammary gland cancer induced by 7,12-dimethylbenzo(a)anthracene. No significant influence on the formation of fibroadenomas as a side effect was reported.


Subject(s)
Histiocytoma, Benign Fibrous/immunology , Histiocytoma, Benign Fibrous/prevention & control , Mammary Neoplasms, Experimental/immunology , Mammary Neoplasms, Experimental/prevention & control , Trichinellosis/immunology , Animals , Female , Histiocytoma, Benign Fibrous/chemically induced , Histiocytoma, Benign Fibrous/microbiology , Male , Mammary Neoplasms, Experimental/chemically induced , Mammary Neoplasms, Experimental/microbiology , Rats
17.
Anticancer Res ; 22(6A): 3261-6, 2002.
Article in English | MEDLINE | ID: mdl-12530073

ABSTRACT

BACKGROUND: Attenuated Salmonella typhimurium has been demonstrated as a potential gene delivery vector. Previous findings induce the necessity to optimize tumor selectivity and bacterial dosing in relation to tumor volume and intratumoral therapeutic gene expression. MATERIALS AND METHODS: Attenuated Salmonella VNP20009 and VNP20047 (expressing cytosine deaminase) were systemically administered to tumor-bearing rats. The bacteria were quantified in tumor and normal organs. Conversion of 5-fluorocytosine to 5-fluorouracil was evaluated using thin layer chromatography. RESULTS: Tumor colonization efficiency was dependent on Salmonella density, administration route and tumor volume. Colonization of normal tissues gradually decreased with time, while intratumoral proliferation of bacteria remained high during the follow-up period. The Optimal Therapeutic Dose (OTD) was found to be 5.10(7) cfu/rat. Intratumoral VNP20047-expressed CDase leading to the conversion of 5-FC to 5-FU was detected in vivo. CONCLUSION: Our results indicate the need to define an OTD, probably for each species, when using genetically engineered Salmonella as a tumor- and species-selective vector in cancer therapy.


Subject(s)
Genetic Therapy/methods , Mammary Neoplasms, Experimental/microbiology , Mammary Neoplasms, Experimental/therapy , Rhabdomyosarcoma/microbiology , Rhabdomyosarcoma/therapy , Salmonella typhimurium/genetics , Animals , Antimetabolites, Antineoplastic/pharmacokinetics , Cytosine Deaminase , Flucytosine/pharmacokinetics , Fluorouracil/pharmacokinetics , Genetic Vectors/genetics , Genetic Vectors/metabolism , Male , Mammary Neoplasms, Experimental/drug therapy , Neoplasm Transplantation , Nucleoside Deaminases/genetics , Nucleoside Deaminases/metabolism , Prodrugs/pharmacokinetics , Rats , Rats, Inbred F344 , Rhabdomyosarcoma/drug therapy , Salmonella typhimurium/enzymology , Salmonella typhimurium/metabolism , Tissue Distribution
18.
J Virol ; 68(8): 5019-26, 1994 Aug.
Article in English | MEDLINE | ID: mdl-8035502

ABSTRACT

Mouse mammary tumor virus is a replication-competent B-type murine retrovirus responsible for mammary gland tumorigenesis in some strains of laboratory mice. Mouse mammary tumor virus is transmitted horizontally through the milk (exogenous or milk-borne virus) to susceptible offspring or vertically through the germ line (endogenous provirus). Exogenously acquired and some endogenous mouse mammary tumor viruses are expressed at high levels in lactating mammary glands. We show here that there is packaging of the endogenous Mtv-1 virus, which is expressed at high levels in the lactating mammary glands of C3H/HeN mice, by the virions of exogenous C3H mouse mammary tumor virus [MMTV(C3H)]. The mammary tumors induced in C3H/HeN mice infected with exogenous MMTV (C3H) virus contained integrated copies of recombinant virus containing a region of the env gene from an endogenous virus. This finding indicates that there was copackaging of the Mtv-1 and MMTV(C3H) RNAs in the same virions. Moreover, because Mtv-1 encodes a superantigen protein with a V beta specificity different from that encoded by the exogenous virus, the packaging of Mtv-1 results in an infectious virus with a broader host range than MMTV(C3H).


Subject(s)
Mammary Tumor Virus, Mouse/genetics , RNA, Viral/biosynthesis , Recombination, Genetic , Animals , Base Sequence , DNA, Viral , Female , Lactation , Male , Mammary Neoplasms, Experimental/microbiology , Mammary Tumor Virus, Mouse/physiology , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Molecular Sequence Data , Proviruses/physiology , Retroviridae Infections/microbiology , Retroviridae Infections/transmission , Tumor Virus Infections/microbiology , Tumor Virus Infections/transmission , Virus Replication
19.
Cancer Lett ; 79(2): 199-211, 1994 May 16.
Article in English | MEDLINE | ID: mdl-8019980

ABSTRACT

The effects of caloric restriction on the incidence of mammary tumor development, the levels of the expression of mouse mammary tumor virus (MMTV)- and prolactin-RNA, as well as the levels of serum prolactin, were investigated in virgin RIII/Sa mice, a strain known to display a high incidence of spontaneous mammary tumor development. Of the 54 mice fed a low-calorie (LC; 10 kcal/day) diet containing low fat (LF; 5% corn oil) for a period of 72 weeks, only seven mice were found to develop mammary tumors, an incidence of 13%. By contrast, the cumulative tumor incidence in a similar sized group of mice, fed a high-calorie (HC; 16 kcal/day) low fat-containing diet was 73%. Estimation of the relative levels of MMTV-RNA, as determined by Northern and slot blot hybridizations, in the mammary glands of mice fed LCLF and HCLF diets for 8, 10, 16, 28, and 36 weeks revealed that the LCLF diet-fed mice expressed 4-15-fold less RNA than the HCLF diet-fed mice. Interestingly, however, the LCLF diet did not appear to exert any effect on the expression of prolactin RNA even though it reduced the levels of serum prolactin. We suggest that in RII/Sa mice the modulation of MMTV-induced mammary tumors by dietary calorie is linked to the secretion of serum prolactin which, in turn, affects the replication of MMTV required for mammary cell transformation.


Subject(s)
Diet , Dietary Fats/administration & dosage , Energy Intake , Mammary Neoplasms, Experimental/etiology , Animals , Blotting, Northern , Body Weight , Female , Incidence , Mammary Neoplasms, Experimental/epidemiology , Mammary Neoplasms, Experimental/genetics , Mammary Neoplasms, Experimental/microbiology , Mammary Tumor Virus, Mouse/isolation & purification , Mice , Prolactin/blood , Prolactin/genetics , RNA, Viral/isolation & purification , Random Allocation
20.
Cancer Res ; 54(6): 1529-35, 1994 Mar 15.
Article in English | MEDLINE | ID: mdl-8137259

ABSTRACT

Lymphocytic infiltrates of BALB/c C4 hyperplastic alveolar nodule (HAN) have elevated natural killer (NK) activity, which correlates positively with the progression of C4 HAN to tumor: C4 HAN produces an infectious mouse mammary tumor virus, MMTV(C4), which encodes a superantigen that activates and deletes T-cells with the V beta 2 segment in the T-cell receptor. In this report, NK activation by both MMTV(C4) and MMTV(C4) superantigen was tested. NK activity was measured in naive BALB/c mice, BALB/c mice depleted of V beta 2+ T-cell, or V beta 2-transgenic mice after they received injections of either purified MMTV(C4) or MMTV(C4)-infected splenocytes. Elevated NK activity was observed in BALB/c mice receiving MMTV(C4) or MMTV(C4)-infected splenocytes. Depletion of V beta 2+, but not V beta 8+, T-cells by specific anti-V beta hybridoma before injection of MMTV(C4)-infected cells reduced but did not eliminate NK activation. NK activation in V beta 2-transgenic mice occurred before massive CD4 T-cell deletion took place and was more pronounced than that in the nontransgenic littermates. These results indicate that MMTV activates NK cells through superantigen-dependent and -independent pathways and supports the role of MMTV(C4) in the augmented NK activity observed in C4 HAN infiltrates. The progression of C4 HAN to tumor represents a model system for the analysis of how tumorigenesis may be affected by lesion-associated viruses.


Subject(s)
Killer Cells, Natural/immunology , Killer Cells, Natural/microbiology , Lymphocyte Activation/immunology , Mammary Tumor Virus, Mouse , Receptors, Antigen, T-Cell, alpha-beta/genetics , Receptors, Antigen, T-Cell, alpha-beta/physiology , Animals , Base Sequence , Female , Hyperplasia/microbiology , Killer Cells, Natural/drug effects , Lymphocyte Activation/drug effects , Lymphocyte Activation/physiology , Lymphocyte Depletion , Male , Mammary Neoplasms, Experimental/microbiology , Mammary Tumor Virus, Mouse/immunology , Mice , Mice, Inbred BALB C , Mice, Transgenic , Molecular Sequence Data , Pulmonary Alveoli/microbiology , Pulmonary Alveoli/pathology , Retroviridae Infections/immunology , Retroviridae Infections/microbiology , Spleen/cytology , Spleen/metabolism , Superantigens/immunology , Superantigens/pharmacology , T-Lymphocytes/cytology , T-Lymphocytes/immunology , Tumor Virus Infections/immunology , Tumor Virus Infections/microbiology
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