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      1. Author :
        Jenkins, Darlene E; Hornig, Yvette S; Oei, Yoko; Dusich, Joan; Purchio, Tony
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Breast cancer research: BCR
      6. Products :
      7. Volume :
        7
      8. Issue :
        4
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Breast Neoplasms; Disease Models, Animal; Female; Humans; Luciferases; Mammary Neoplasms, Animal; MDA-MB-231-D3H2LN cells; Mice; Mice, Nude; Neoplasm Metastasis; Plasmids; Transplantation, Heterologous; Tumor Cells, Cultured
      12. Abstract :
        INTRODUCTION Our goal was to generate xenograft mouse models of human breast cancer based on luciferase-expressing MDA-MB-231 tumor cells that would provide rapid mammary tumor growth; produce metastasis to clinically relevant tissues such as lymph nodes, lung, and bone; and permit sensitive in vivo detection of both primary and secondary tumor sites by bioluminescent imaging. METHOD Two clonal cell sublines of human MDA-MB-231 cells that stably expressed firefly luciferase were isolated following transfection of the parental cells with luciferase cDNA. Each subline was passaged once or twice in vivo to enhance primary tumor growth and to increase metastasis. The resulting luciferase-expressing D3H1 and D3H2LN cells were analyzed for long-term bioluminescent stability, primary tumor growth, and distal metastasis to lymph nodes, lungs, bone and soft tissues by bioluminescent imaging. Cells were injected into the mammary fat pad of nude and nude-beige mice or were delivered systemically via intracardiac injection. Metastasis was also evaluated by ex vivo imaging and histologic analysis postmortem. RESULTS The D3H1 and D3H2LN cell lines exhibited long-term stable luciferase expression for up to 4-6 months of accumulative tumor growth time in vivo. Bioluminescent imaging quantified primary mammary fat pad tumor development and detected early spontaneous lymph node metastasis in vivo. Increased frequency of spontaneous lymph node metastasis was observed with D3H2LN tumors as compared with D3H1 tumors. With postmortem ex vivo imaging, we detected additional lung micrometastasis in mice with D3H2LN mammary tumors. Subsequent histologic evaluation of tissue sections from lymph nodes and lung lobes confirmed spontaneous tumor metastasis at these sites. Following intracardiac injection of the MDA-MB-231-luc tumor cells, early metastasis to skeletal tissues, lymph nodes, brain and various visceral organs was detected. Weekly in vivo imaging data permitted longitudinal analysis of metastasis at multiple sites simultaneously. Ex vivo imaging data from sampled tissues verified both skeletal and multiple soft tissue tumor metastasis. CONCLUSION This study characterized two new bioluminescent MDA-MB-231-luc human breast carcinoma cell lines with enhanced tumor growth and widespread metastasis in mice. Their application to current xenograft models of breast cancer offers rapid and highly sensitive detection options for preclinical assessment of anticancer therapies in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15987449
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8960
      1. Author :
        Takeshita, Fumitaka; Minakuchi, Yoshiko; Nagahara, Shunji; Honma, Kimi; Sasaki, Hideo; Hirai, Kotaro; Teratani, Takumi; Namatame, Nachi; Yamamoto, Yusuke; Hanai, Koji; Kato, Takashi; Sano, Akihiko; Ochiya, Takahiro
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Proceedings of the National Academy of Sciences of the United States of America
      6. Products :
      7. Volume :
        102
      8. Issue :
        34
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Bone Neoplasms; Cell Line, Tumor; Collagen; DNA-Binding Proteins; Drug Carriers; Gene Expression Regulation, Neoplastic; Gene Therapy; Humans; Luciferases; Male; Mice; PC-3M-luc; Phosphatidylinositol 3-Kinases; Prostatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; Transcription Factors
      12. Abstract :
        Silencing of gene expression by small interfering RNAs (siRNAs) is rapidly becoming a powerful tool for genetic analysis and represents a potential strategy for therapeutic product development. However, there are no reports of systemic delivery for siRNAs toward treatment of bone-metastatic cancer. Accordingly, we report here that i.v. injection of GL3 luciferase siRNA complexed with atelocollagen showed effective reduction of luciferase expression from bone-metastatic prostate tumor cells developed in mouse thorax, jaws, and/or legs. We also show that the siRNA/atelocollagen complex can be efficiently delivered to tumors 24 h after injection and can exist intact at least for 3 days. Furthermore, atelocollagen-mediated systemic administration of siRNAs such as enhancer of zeste homolog 2 and phosphoinositide 3'-hydroxykinase p110-alpha-subunit, which were selected as candidate targets for inhibition of bone metastasis, resulted in an efficient inhibition of metastatic tumor growth in bone tissues. In addition, upregulation of serum IL-12 and IFN-alpha levels was not associated with the in vivo administration of the siRNA/atelocollagen complex. Thus, for treatment of bone metastasis of prostate cancer, an atelocollagen-mediated systemic delivery method could be a reliable and safe approach to the achievement of maximal function of siRNA.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/16091473
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8979
      1. Author :
        Dernell, William S.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        N/A
      6. Products :
      7. Volume :
        N/A
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        *Breast Cancer; *Chemotherapy; *Genes; *Luciferase; Anatomy and Physiology; Biochemistry; Bioware; Cells(Biology); Diseases; Drugs; Efficacy; Gel Polymers; Gels; Growth(Physiology); Humans; Image Processing; In Vitro Analysis.; In Vivo Analysis; Luciferase Genes; Medicine and Medical Research; Metastasis; Mouse Models; Paclitaxel Sensitivity; Poloxamer Polymers; Polymers; Preclinical Evaluations; surgery; Synergism; Toxicity; Tumor Cell Lines
      12. Abstract :
        This project evaluated paclitaxel chemotherapy delivery from a gel polymer system placed into a wound bed following conservative (marginal) surgical removal of human breast cancers grown in nude mice. This delivery method was shown to control local tumor disease as well as assist in control of systemic metastasis. We established 5 human breast cancer cell lines within our laboratory. We elected purchase and implement a unique (luciferase) imaging system which allows in vivo imaging of tumor growth and metastasis (and subsequently decrease animal use). Tumor cell lines were transfected with the luciferase gene. In vitro testing of cell lines established paclitaxel sensitivity and showed a synergistic effect of delivering paclitaxel by the poloxamer polymer, especially for the chemotherapy resistant cell line, MCF-7-ADR. We completed the simultaneous evaluation of local and systemic toxicity, local, regional and systemic distribution and local and systemic efficacy of locally delivered paclitaxel chemotherapy following tumor removal using the MCF-7-ADR cell line in nude mice. Intracavitary administration of taxol in poloxamer was well tolerated (locally and systemically) afld resulted in significantly improved control of local tumor regrowth and comparable control of metastasis following marginal tumor removal as compared to intravenous paclitaxel (parent drug) . Sustained drug levels (from polymer delivery) were seen in plasma and liver tissue at 60 days.
      13. URL :
        http://oai.dtic.mil/oai/oai?verb=getRecord&metadataPrefix=html&identifier=ADA437225
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8994
      1. Author :
        Jenkins, Darlene E; Hornig, Yvette S; Oei, Yoko; Dusich, Joan; Purchio, Tony
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Breast cancer research: BCR
      6. Products :
      7. Volume :
        7
      8. Issue :
        4
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Breast Neoplasms; Disease Models, Animal; Female; Humans; Luciferases; Mammary Neoplasms, Animal; MDA-MB-231-D3H1 cells; Mice; Mice, Nude; Neoplasm Metastasis; Plasmids; Transplantation, Heterologous; Tumor Cells, Cultured
      12. Abstract :
        INTRODUCTION Our goal was to generate xenograft mouse models of human breast cancer based on luciferase-expressing MDA-MB-231 tumor cells that would provide rapid mammary tumor growth; produce metastasis to clinically relevant tissues such as lymph nodes, lung, and bone; and permit sensitive in vivo detection of both primary and secondary tumor sites by bioluminescent imaging. METHOD Two clonal cell sublines of human MDA-MB-231 cells that stably expressed firefly luciferase were isolated following transfection of the parental cells with luciferase cDNA. Each subline was passaged once or twice in vivo to enhance primary tumor growth and to increase metastasis. The resulting luciferase-expressing D3H1 and D3H2LN cells were analyzed for long-term bioluminescent stability, primary tumor growth, and distal metastasis to lymph nodes, lungs, bone and soft tissues by bioluminescent imaging. Cells were injected into the mammary fat pad of nude and nude-beige mice or were delivered systemically via intracardiac injection. Metastasis was also evaluated by ex vivo imaging and histologic analysis postmortem. RESULTS The D3H1 and D3H2LN cell lines exhibited long-term stable luciferase expression for up to 4-6 months of accumulative tumor growth time in vivo. Bioluminescent imaging quantified primary mammary fat pad tumor development and detected early spontaneous lymph node metastasis in vivo. Increased frequency of spontaneous lymph node metastasis was observed with D3H2LN tumors as compared with D3H1 tumors. With postmortem ex vivo imaging, we detected additional lung micrometastasis in mice with D3H2LN mammary tumors. Subsequent histologic evaluation of tissue sections from lymph nodes and lung lobes confirmed spontaneous tumor metastasis at these sites. Following intracardiac injection of the MDA-MB-231-luc tumor cells, early metastasis to skeletal tissues, lymph nodes, brain and various visceral organs was detected. Weekly in vivo imaging data permitted longitudinal analysis of metastasis at multiple sites simultaneously. Ex vivo imaging data from sampled tissues verified both skeletal and multiple soft tissue tumor metastasis. CONCLUSION This study characterized two new bioluminescent MDA-MB-231-luc human breast carcinoma cell lines with enhanced tumor growth and widespread metastasis in mice. Their application to current xenograft models of breast cancer offers rapid and highly sensitive detection options for preclinical assessment of anticancer therapies in vivo.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15987449
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8993
      1. Author :
        Hunter, John J.; Neben, Tamlyn Yee; Purchio, Tony; Jenkins, Darlene
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        AACR Meeting Abstracts
      6. Products :
      7. Volume :
        2005
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Bioware; SKOV3-luc-D3 cells
      12. Abstract :
        Peritoneal dissemination is a common feature of human ovarian carcinoma. While this can be mimicked in preclinical models by intraperitoneal injection of human ovarian tumor cells into immunocompromised mice, the resulting tumor burden is difficult to monitor and quantify. Intraperitoneal tumor growth is typically evaluated indirectly by measured changes in mouse abdominal girth and body weight or, directly, by macroscopic and histological examination at the endpoint of the study. In order to establish a model system that allows continuous and accurate assessment of ovarian cancer growth and spread over time we transfected SKOV-3 cells with the firefly luciferase gene. The resulting cell line, SKOV3-luc-D3, expresses stable levels of luciferase in vitro and emits a strong luminescent signal when exposed to luciferin. Xenograft tumors established with this cell line can be tracked and quantified non-invasively by bioluminescent imaging using a highly sensitive, cooled CCD camera (IVIS(R) Imaging System, Xenogen Corp). In addition to providing a direct measure of primary tumor burden and growth, the SKOV3-luc-D3 cell line also allows for real-time evaluation of tumor response to various therapeutic agents, as well as enhanced detection of distal metastases.
      13. URL :
        http://www.aacrmeetingabstracts.org/cgi/content/abstract/2005/1/256-b
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9014
      1. Author :
        Neben, Tamlyn Yee; Clermont, Anne O.; Esposito, Lin; Oei, Yoko; Neben, Tamlyn Yee; Jenkins, Darlene E.; Clermont, Anne O.; Esposito, Lin; Oei, Yoko; Jenkins, Darlene E.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        AACR Meeting Abstracts
      6. Products :
      7. Volume :
        2005
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Bioware; Lovo-6-luc-1 cells
      12. Abstract :
        Colorectal cancer is the fourth most common cancer in the United States with an estimated 130,000 new cases diagnosed each year. Many cases are asymptomatic and not diagnosed until late stage of disease. Identification of primary tumors at an earlier stage is advantageous in treatment planning and aids in decreasing the morbidity/mortality rate from recurrence. The aim of our studies is to establish a xenograft system for monitoring tumor growth and metastasis in vivo which allows continual evaluation of drug and drug regimen efficacy at all stages of tumor progression. LoVo-6-luc-1, a luciferase expressing cell line derived from LoVo human colorectal adenocarcinoma cells, was injected by various routes (subcutaneous, intraperitoneal and intracecal) into female SCID-bg mice. Tumor growth and metastatic spread was monitored weekly by in vivo imaging using the Xenogen IVISTM imaging platform. Visible bioluminescence signals were detected immediately after injection and high tumor take was seen in all of the models. In the subcutaneous model, we found a high correlation between mean bioluminescence and mean tumor volume. In the intraperitoneal and ceacum injected models, the onset of tumor spread was rapid and ex vivo imaging confirmed metastasis to multiple organs such as liver, lung, kidney, adrenal gland, spleen and ovary.
      13. URL :
        http://www.aacrmeetingabstracts.org/cgi/content/abstract/2005/1/908-d
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9016
      1. Author :
        Xiong, Y. Q.; Willard, J.; Kadurugamuwa, J. L.; Yu, J.; Francis, K. P.; Bayer, A. S.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Antimicrobial Agents and Chemotherapy
      6. Products :
      7. Volume :
        49
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen; Bioware; Xen29
      12. Abstract :
        Therapeutic options for invasive Staphylococcus aureus infections have become limited due to rising antimicrobial resistance, making relevant animal model testing of new candidate agents more crucial than ever. In the present studies, a rat model of aortic infective endocarditis (IE) caused by a bioluminescently engineered, biofilm-positive S. aureus strain was used to evaluate real-time antibiotic efficacy directly. This strain was vancomycin and cefazolin susceptible but gentamicin resistant. Bioluminescence was detected and quantified daily in antibiotic-treated and control animals with IE, using a highly sensitive in vivo imaging system (IVIS). Persistent and increasing cardiac bioluminescent signals (BLS) were observed in untreated animals. Three days of vancomycin therapy caused significant reductions in both cardiac BLS (>10-fold versus control) and S. aureus densities in cardiac vegetations (P < 0.005 versus control). However, 3 days after discontinuation of vancomycin therapy, a greater than threefold increase in cardiac BLS was observed, indicating relapsing IE (which was confirmed by quantitative culture). Cefazolin resulted in modest decreases in cardiac BLS and bacterial densities. These microbiologic and cardiac BLS differences during therapy correlated with a longer time-above-MIC for vancomycin (>12 h) than for cefazolin (?4 h). Gentamicin caused neither a reduction in cardiac S. aureus densities nor a reduction in BLS. There were significant correlations between cardiac BLS and S. aureus densities in vegetations in all treatment groups. These data suggest that bioluminescent imaging provides a substantial advance in the real-time monitoring of the efficacy of therapy of invasive S. aureus infections in live animals.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15743898
      14. Call Number :
        144577
      15. Serial :
        7474
      1. Author :
        Yu, Jun; Wu, Jenny; Francis, Kevin P; Purchio, Tony F; Kadurugamuwa, Jagath L
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        The Journal of antimicrobial chemotherapy
      6. Products :
      7. Volume :
        55
      8. Issue :
        4
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Anti-Bacterial Agents; Biofilms; Bioware; Drug Resistance, Bacterial; Mice; Mutation; Rifampin; Staphylococcus aureus; Xen29
      12. Abstract :
        OBJECTIVES To investigate in vivo fitness of rifampicin-resistant Staphylococcus aureus mutants in a mouse biofilm model using bioluminescence imaging. MATERIALS AND METHODS S. aureus was engineered with a luciferase operon to emit bioluminescence that can be detected in vivo using an IVIS imaging system. Two rifampicin-resistant strains of S. aureus that were previously isolated from animals undergoing rifampicin treatment, S464P (resistant to low concentrations of rifampicin) and H481Y (resistant to high concentrations of rifampicin), were characterized and then compared with their parental strain for in vivo fitness to form biofilm infections in the absence of rifampicin. RESULTS The mutant S464P showed better adaptation to in vivo growth than either the parental strain or H481Y without selective pressure. Six days after implanting pre-colonized catheters, bioluminescent signals were seen from 100% of the catheters coated by the mutant S464P. In comparison, only 83% and 61% of the catheters coated by the parental strain and H481Y, respectively, maintained a signal in vivo. Rifampicin treatment of S464P biofilms in vivo resulted in a slight decline, but earlier rebound in bioluminescence from these catheters compared with the parental signal, whereas rifampicin had no affect on bioluminescence in mice infected with mutant H481Y. CONCLUSIONS The mutant with low-level rifampicin resistance appears to be better adapted to in vivo growth than the mutant that has high-level rifampicin resistance. Moreover, the former mutant may actually have a slight competitive advantage over the rifampicin-susceptible strain (parental), raising awareness for the occurrence of such strains in clinical environments.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/15743898
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9055
      1. Author :
        Georgel, Philippe; Crozat, Karine; Lauth, Xavier; Makrantonaki, Evgenia; Seltmann, Holger; Sovath, Sosathya; Hoebe, Kasper; Du, Xin; Rutschmann, Sophie; Jiang, Zhengfan; Bigby, Timothy; Nizet, Victor; Zouboulis, Christos C; Beutler, Bruce
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2005
      5. Publication :
        Infection and immunity
      6. Products :
      7. Volume :
        73
      8. Issue :
        8
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Anti-Bacterial Agents; Bioware; Chromosome Mapping; Eye Diseases; Fatty Acids, Monounsaturated; Likelihood Functions; Lod Score; Mice; Mice, Inbred C57BL; Oleic Acid; Receptors, Immunologic; Sequence Analysis, DNA; Skin; Staphylococcal Skin Infections; Stearoyl-CoA Desaturase; Streptococcus pyogenes; Time Factors; Toll-Like Receptor 2; Xen8.1, Xen20, Xen14
      12. Abstract :
        flake (flk), an N-ethyl-N-nitrosourea-induced recessive germ line mutation of C57BL/6 mice, impairs the clearance of skin infections by Streptococcus pyogenes and Staphylococcus aureus, gram-positive pathogens that elicit innate immune responses by activating Toll-like receptor 2 (TLR2). Positional cloning and sequencing revealed that flk is a novel allele of the stearoyl coenzyme A desaturase 1 gene (Scd1). flake homozygotes show reduced sebum production and are unable to synthesize the monounsaturated fatty acids (MUFA) palmitoleate (C(16:1)) and oleate (C(18:1)), both of which are bactericidal against gram-positive (but not gram-negative) organisms in vitro. However, intradermal MUFA administration to S. aureus-infected mice partially rescues the flake phenotype, which indicates that an additional component of the sebum may be required to improve bacterial clearance. In normal mice, transcription of Scd1-a gene with numerous NF-kappaB elements in its promoter--is strongly and specifically induced by TLR2 signaling. Similarly, the SCD1 gene is induced by TLR2 signaling in a human sebocyte cell line. These observations reveal the existence of a regulated, lipid-based antimicrobial effector pathway in mammals and suggest new approaches to the treatment or prevention of infections with gram-positive bacteria.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/16040962
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9990
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