Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases

Yang, Meng; Baranov, Eugene; Jiang, Ping; Sun, Fangxian; Li, Xiaoming; Li, Lingna; Hasegawa, S.; Bouvet, Michael; Al-Tuwaijri, Maraya; Chishima, Takashi; Shimada, Hiroshi; Moossa, A. R.; Penman, Sheldon; Hoffman, Robert M.

doi:10.1073/pnas.97.3.1206

Cited by 460 publications

(305 citation statements)

References 20 publications

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“…The usage of GFP-expressing cancer cells permits the detection and visualization of distant micrometastases in fresh organ. 8 Yang et al 12 quantitated metastatic tumor growth by observation of external GFP images. In our study, we observed micrometastases (30 m in size) in whole lung or lymph nodes with thicknesses of 1 to 3 mm.…”

Section: Discussionmentioning

confidence: 99%

Anti‐metastatic effect of capecitabine on human colon cancer xenografts in nude mouse rectum

Ninomiya

Terada

Yoshizumi

et al. 2004

Intl Journal of Cancer

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Key words: capecitabine; lymph node metastasis; lung metastasis; thymidine phosphorylase; rectal xenograft modelGastrointestinal cancer is one of the most common internal malignancies and is one of the leading causes of cancer-related morbidity and mortality in the world. The primary definitive treatment modality is surgical excision. Adjuvant therapies subsequent to surgery have been studied extensively in recent years because of the high incidence of postoperative recurrences. More than 40 years after its development, 5-FU remains the chemotherapeutic mainstay for the management of patients with many types of cancer. In recent years, several new approaches to the treatment of cancer have undergone examination. These include the introduction of 2 novel cytotoxic compounds. Another strategy that has received attention has been the development of several oral forms of fluoropyrimidine, offering an alternative route of administration to intravenous 5-FU 1 . Capecitabine (N 4 -pentyloxycarbonyl-5Ј-deoxy-5-fluorocytidine) is a new fluoropyrimidine carbamate, which is designed to be activated to 5-FU by 3 sequential steps of enzyme reactions. 1 In humans, it is sequentially converted first to 5Ј-deoxy-5-fluorocytidine by carboxylesterase located in the liver, then to 5Ј-deoxy-5-fluorouridine by cytidine deaminase also with high activity in the liver and in various solid tumors and finally to 5-FU by thymidine phosphorylase (dThdPase) with high activity in many types of tumors. [1][2][3] This oral fluoropyrimidine has an advantage over parental therapy with 5-FU in its potential for affording patients greater convenience and comfort. 4 -6 Randomized clinical trials that compared capecitabine with parenteral 5-FU/leucovorin against metastatic colorectal cancer demonstrated that capecitabine provides at least equivalence, a favorable benefit-risk ratio and greater patient preference making it an acceptable replacement for 5-FU/leucovorin. 4,5,7 Our interest focused on the possibility of capecitabine usage in the prevention of metastasis. Neoadjuvant or adjuvant chemotherapy to inhibit spontaneous metastasis or suppress occult micrometastasis may reduce the incidence of postoperative distant metastasis and improve survival. To confirm this possibility, large randomized clinical trials should be needed. These clinical trials are expensive and time consuming. Animal experiments may take their place to examine the efficacy of new therapeutic modalities. Here, we develop an animal model of gastrointestinal cancer by the intrarectal injection of tumor cells, which resulted in the formation of local tumors with lymph node and lung metastasis consistent with the progression of gastrointestinal cancer in humans.In our study, we examined the efficacy of capecitabine for the inhibition of metastasis and its mechanism for doing so by using a mouse metastasis model. MATERIAL AND METHODS AnimalsMale BALB/c-nu/nu mice aged 5 weeks were obtained from Charles River Japan, Inc. (Kanagawa, Japan). After at least 1 week of observation, th...

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Section: Discussionmentioning

confidence: 99%

Anti‐metastatic effect of capecitabine on human colon cancer xenografts in nude mouse rectum

Ninomiya

Terada

Yoshizumi

et al. 2004

Intl Journal of Cancer

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“…Also human-specific surface markers like HLA-DR or human endogenous retroviral sequences (Ailles et al, 1999) are used. Others, artificially transfected cells with marker genes like lac Z (Hayashi et al, 1996;Krüger et al, 1999) or green fluorescent protein (Yang et al, 2000) for a detection of the specific gene product in the in vivo xenotransplantation system.…”

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confidence: 99%

Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems

et al. 2002

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The sensitive detection of human cells in immunodeficient rodents is a prerequisite for the monitoring of micrometastasis of solid tumours, dissemination of leukaemic cells, or engraftment of haematological cells. We developed a universally applicable polymerase chain reaction method for the detection of a human-specific 850-bp fragment of the a-satellite DNA on human chromosome 17. The method allows the detection of one human cell in 10 6 murine cells and could be established as both, a conventional DNA polymerase chain reaction-assay for routine screening, and a quantitative real-time polymerase chain reaction-assay using TaqMan-methodology. It was applied to the following xenotransplantation systems in SCID and NOD/ SCID mice: (1) In a limiting dilution assay, cells of the MDA-MB 435 breast carcinoma were injected into the mammary fat pad of NOD/SCID mice. It could be shown that 10 cells mouse 71 were sufficient to induce a positive polymerase chain reaction signal in liver and lung tissue 30 days after transplantation as an indicator for micrometastasis. At this time a palpable tumour was not yet detectable in the mammary fat pad region. (2) Cells of a newly established human acute lymphatic leukaemia were administered intraperitoneally to SCID mice. These cells apparently disseminated and were detectable as early as day 50 in the peripheral blood of living mice, while the leukaemia manifestation was delayed by day 140. (3) In a transplantation experiment using mature human lymphocytes we wanted to standardise conditions for a successful survival of these cells in NOD/SCID mice. It was established that at least 5610 7 cells given intravenously were necessary and that the mice had to be conditioned by 2 Gy body irradiation to get positive polymerase chain reaction bands in several organs. (4) Engraftment studies with blood stem cells originating from cytapheresis samples of tumour patients or from cord blood were undertaken in NOD/SCID mice in order to define conditions of successful engraftment and to use this model for further optimisation strategies. The polymerase chain reaction method presented allowed a reliable prediction of positive engraftment and agreed well with the results of immunohistochemical or FACS analysis. All together, the polymerase chain reaction method developed allows a sensitive and reliable detection of low numbers of human cells in immunodeficient hosts. In combination with real-time (TaqMan) technique it allows an exact quantification of human cells. As this method can be performed with accessible material of living animals, follow up studies for the monitoring of therapeutic interventions are possible in which the survival time of mice as evaluation criteria can be omitted.

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“…NV1066 carries such a marker gene, a constitutively expressed transgene for EGFP, the protein product of which is identifiable 4-6 hours following viral entry into cells. While green fluorescent protein and its derivative proteins have been used extensively as reporters and/or markers in numerous biologic studies (60)(61)(62)(63)(64)(65), their use to determine infection and spread of replication-competent viral vectors in vivo is an emerging technology. Previous studies have demonstrated that the fluorescent signal from cells constitutively expressing GFP correlates strongly with cell number in vitro (66).…”

Section: Discussionmentioning

confidence: 99%

Minimally invasive localization of oncolytic herpes simplex viral therapy of metastatic pleural cancer

et al. 2005

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Purpose-Herpes simplex virus-one (HSV-1) oncolytic therapy and gene therapy are promising treatment modalities against cancer. NV1066, one such HSV-1 virus carries a marker gene for enhanced green fluorescent protein (EGFP). The purpose of this study was to determine whether NV1066 is cytotoxic to lung cancer and whether EGFP is a detectable marker of viral infection in vitro and in vivo. We further investigated whether EGFP expression in infected cells can be used to localize the virus and to identify small metastatic tumor foci (< 1 mm.) in vivo by means of minimally invasive endoscopic systems equipped with fluorescent filters.Experimental Design-In A549 human lung cancer cells, in vitro viral replication was determined by plaque assay, cell kill by LDH release assay, and EGFP expression by flow cytometry. In vivo, A549 cells were injected into the pleural cavity of athymic mice. Mice were treated with intrapleural injection of NV1066 or saline and examined for EGFP expression in tumor deposits using a stereomicroscope or a fluorescent thoracoscopic system. Results-NV1066 replicated in, expressed EGFP in infected cells and killed tumor cells in vitro.In vivo, treatment with intrapleural NV1066 decreased pleural disease burden, as measured by chest wall nodule counts and organ weights. EGFP was easily visualized in tumor deposits, including microscopic foci, by fluorescent thoracoscopy.Conclusions-NV1066 has significant oncolytic activity against a human NSCLC cell line and is effective in limiting the progression of metastatic disease in an in vivo orthotopic model. By incorporating fluorescent filters into endoscopic systems, a minimally-invasive means for diagnosing small metastatic pleural deposits and localization of viral therapy for thoracic malignancies may be developed using the EGFP marker gene inserted in oncolytic herpes simplex viruses.

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Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases

Cited by 460 publications

References 20 publications

Anti‐metastatic effect of capecitabine on human colon cancer xenografts in nude mouse rectum

Anti‐metastatic effect of capecitabine on human colon cancer xenografts in nude mouse rectum

Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems

Minimally invasive localization of oncolytic herpes simplex viral therapy of metastatic pleural cancer

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