FEATURED ARTICLE
Systemic targeting of primary bone tumor and lung metastasis of high-grade osteosarcoma in nude mice with a tumor-selective strain of Salmonella typhymurium.
Hayashi, K., Zhao, M., Yamauchi, K., Yamamoto, N., Tsuchiya, H., Tomita, K., Kishimoto, H., Bouvet, M., and Hoffman, R.M.
Cell Cycle 8, 870-875, 2009.
FEATURED ARTICLE
In vitro/in vivo biorecognition of lectin-immobilized fluorescent nanospheres for human colorectal cancer cells.
Sakuma, S., Yano, T., Masaoka, Y., Kataoka, M., Hiwatari, K., Tachikawa, H., Shoji, Y., Kimura, R., Ma, H., Yang, Z., Tang, L., Hoffman, R.M., Yamashita, S.
Journal of Controlled Release 134, 2-10, 2009.
FEATURED ARTICLE
Glowing proteins with promising biological and medical applications.
by Linda Zajac
Chem Matters 12-14, December
2008.
FEATURED ARTICLE
Dual-color imaging of angiogenesis and its
inhibition in bone and soft tissue sarcoma.
J. Surg. Res. 140, 165-170,
2007.
FEATURED ARTICLE
Tumor cells genetically labeled with GFP in the nucleus and RFP in the cytoplasm for imaging cellular dynamics. T
Jiang, P., Yamauchi, K., Yang., Tsuji, K., Xu, M., Maitra, A., Bouvet, M., and Hoffman, R.M.
Cell Cycle
5, 1198-1201, 2006.
ABSTRACT
Dual-color fluorescent cells with one color fluorescent protein in the nucleus and another color fluorescent protein in the cytoplasm were genetically engineered. The dual-color cancer cells enable real-time nuclear-cytoplasmic dynamics to be visualized in living cells in vivo as well as in vitro. To obtain the dual-color cells, red fluorescent protein (RFP) was expressed in the cytoplasm of a series of human and rodent cancer cells, and green fluorescent protein (GFP) linked to histone H2B was expressed in the nucleus. Nuclear GFP expression enabled visualization of nuclear dynamics, whereas simultaneous cytoplasmic RFP expression enabled visualization of nuclear-cytoplasmic ratios as well as simultaneous cell and nuclear shape changes. Using the Olympus OV100 Whole-Mouse Imaging System, total sub-cellular dynamics can be visualized in the living dual-color cells in real time in the live mouse after cell injection. Highly elongated cancer cells and nuclei in narrow capillaries were visualized where both the nuclei and cytoplasm deform. Both cytoplasm and nuclei were visualized to undergo extreme deformation during extravasation with cytoplasmic processing exiting vessels first and nuclei following along these processes. The dual-color cells described here thus enable the sub-cellular dynamics of cancer celltrafficking to be imaged in the living animal.
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Technical Highlights of the Recent Literature
[BIOTECHNIQUES 36, No. 1 January, 19,
2004]
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The
development of tumor-induced vasculature is essential for both tumor growth
and the formation of metastases. Newly formed blood vessels thus provide
promising potential therapeutic targets. However, the role of host tissues
and their interactions with the tumor in this angiogenesis has remained
unclear. Yang et al. have developed a technique for delineating tumor-induced
host processes by exploiting a dual-color fluorescent system. They
transplanted tumors expressing red fluorescent protein (RFP) into green
fluorescent protein (GFP)-expressing nude mice. All of the tissues from these
transgenic mice fluoresce green, except for their hair and erythrocytes. This
dual-color fluorescence imaging is unique in that it can depict morphological
events during tumor-host interactions at the cellular level in fresh tissue
in addition to whole-body imaging. They visualized macrophages engulfing tumor
cells, dendritic cells contacting tumor cells, and lymphocytes attacking the
tumor. The insights gleaned from this study and others like it should be
important in elucidating the role of host cells in tumor growth, progression,
and invasion. Proceedings of the National Academy of Sciences of the USA
100 (24): 14259-14262 (2003)
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FEATURED ARTICLE
Truncated Galectin-3 Inhibits Tumor Growth and Metastasis in Orthotopic Nude Mouse Model of Human Breast Cancer.
John, C.M., Leffler, H., Kahl-Knutsson, B., Svensson, I., Jarvis, G.A.
[Clinical
Cancer Research 9, 2374-2383, 2003]
Orthotopic breast
cancer xenograft imaged with green fluorescent protein in nude mouse treated
with truncated galectin-3 (galectin-3C). The mammalian lectin galectin-3 has
been suggested to promote cancer by stimulation of cell adhesion and inhibition
of apoptosis reulting in increased tumor growth and metastasis. Galectin-3C, a
fragment of galectin-3 retaining its carbohydrate binding activity but lacking
the cross linking ability and thereby possibly acting as a dominant negative
inhibitor, reduced tumor growth and metastasis and showed no toxic side
effects. For details, see C.M. John et
al., in this issue.
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FEATURED ARTICLE
Green fluorescent protein imaging of tumour growth, metastasis and angiogenesis in mouse models.
Robert M. Hoffman
[Lancet Oncology 3, 546-556, 2002]
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In
this issue:
Green fluorescence protein imaging of tumour growth, metastasis, and
angiogenesis in mouse models by Robert M Hoffman. The figure below shows a
whole-body fluorescent image of a mouse with bone metastases in the (a)
skull, (b) scapula, and (c) spine.
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PAPER
Dissecting p53 tumor suppressor functions in vivo
Clemens A. Schmitt, Jordan S. Fridman, Meng Yang, Eugene Baranov, Robert M. Hoffman, and Scott W. Lowe
[CANCER CELL 1, 289-298 April 2002]
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Whole body fluorescence imaging of lymphoma progression in live mice. The cover shows the temporal and spatial progression of Em-myc lymphoma cells tagged with green fluorescent protein in a live mouse (with time progression from top to bottom). Note that the lymphomas first expand within the lympoid compartments and bone. In the absence of p53 or following Bcl-2 overexpression, these lympomas readily disseminate into nonlympoid compartments. For details see Schmitt et al. (pp. 289-298) in this issue. |
Summary:
Although the p53 tumor supressor acts in plethora of processes that influence
cellular proliferation and survival, it remains unclear which p53 functions
are essential for tumor suppression and, as a consequence, are selected
against during tumor development. Using a mouse model harbouring primary,
genetically modified myc-driven lympomas, we show that disruption of apoptosis
downstream of p53 by Bcl2 or a dominant-negative caspase 9 confers-like p53
loss-a selective advantage, and completely alleviates pressure to inactivate
p53 during lymphomagenesis. Despite their p53-null-like aggressive phenotype,
apoptosis-defective lymphomas that retain intact p53 genes do not display the
checkpoint defects and gross aneuploidy that are charcteristic of p53 mutant
tumors. Therefore, apoptosis is the only p53 function selected against during
lymphoma development, whereas defective cell-cycle checkpoints and aneuploidy
are mere byproducts of p53 loss. Figure 4. Whole body fluorescence imaging
allows visualisation of lymphoma dissemination.
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A Senescence Program Contolled by p53 and p16INK4a Contribures to the Outcome of Cancer Therapy
Clemens A. Schmitt, Jordan S. Fridman, Meng Yang, Soyoung Lee, Eugene Baranov, Robert M. Hoffman, and Scott W. Lowe
[CELL 109, 335-346, May 3, 2002]
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Summary p53 andINK4a/ARF mutations promote tumorogenesis and drug resistance, in part, by disabling apoptosis. We show that primary murine lympomas also respond to chemotherapy by engaging a senescence program controlled by p53 and p16INK4a. Hence, tumors with p53 or INK4a/ARF mutations-but not those lacking ARF alone - respond poorly to cyclophosphamide therapy in vivo. Morover, tumors harboring a Bcl2-mediated apoptotic block undergo a drug-induced cytostasis involving the accumulation of p53, p16INK4a, and senescence markers, and typically acquire p53 or INK4a mutations upon progression to a terminal stage. Finally, mice bearing tumors capable of drug-induced senescence have a much better prognosis following chemotherapy than those harboring tumors with senescence defects. Therefore, cellular senescence contributes to treatment outcome in vivo. Figure 1. Contribution of p53 and Bcl2 to Treatment Responses
Mice harboring ctrl.-MSCV, p53 null-MSCV, and ctrl.-bcl2 lymphomas were treated at comparable tumor burdens (day 0) with a single dose of cyclophosphamide (CTX) and monitored by whole-body fluorescence imaging. Representative examples are shown. |
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REVIEW
GFP-Expressing Metastatic-Cancer Mouse Models.
By Robert M. Hoffman
[In: Teicher, B.,
ed. Tumor Models in Cancer Research.
Totowa, NJ: Humana Press Inc., 99-112, 2002].
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REVIEW
Green Fluorescent Protein Imaging of Tumor Cells in Mice
By Robert M. Hoffman
[LAB ANIMAL 31, No. 4, 34-41 April 2002]
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Real-Time Optical Imaging of Primary Tumor Growth and Multiple Metastatic Events in a Pancreatic Cancer Orthotopic Model
Michael Bouvet, Jinwei Wang, Stephanie R. Nardin, Rounak Nassirpour, Meng Yang, Eugene Baranov, Ping Jang, A. R. Moosa and Robert M. Hoffman
[CANCER RESEARCH 62, 1534-1540, March 1,
2002]
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ABSTRACT: We report here whole-body
optical imaging, in real time, of genetically fluorescent pancreatic tumors
growing and metastasizing to multiple sites in live mice. The whole-body
optical imaging system is external and noninvasive. Human pancreatic tumor
cell lines, BxPC-3 and MiaPaCa-2, were engineered to stably express
high-levels of the Acqurea victoria green fluorescent protein (GFP).
The GFP-expressing pancreatic tumor cell lines were surgically orthotopically
implanted as tissue fragments in the body of the pancreas of nude mice.
Whole-body optical images visualized real-time primary tumor growth and
formation of metastatic lesions that developed in the spleen, bowel, portal
lymph nodes, omentum, and liver. Intravital imaging was used for
quatification of growth of micrometastasis on the liver and stomach.
Whole-body imaging was carried out with either a trans-illuminated
epi-fluorescence microscope or a fluorescent light box, both with a
thermoelectrically cooled color CCD camera. The simple, noninvasive, and
highly selective imaging made possible by the strong GFP fluorescence allowed
detailed simultaneous quantitative imaging of tumor growth and multiple
metastasis formation of pancreatic cancer. The GFP imaging affords
unprecendented contionuous visual monitoring of malignant growth and spread
within intact animals without the need for anesthesia, substrate injection,
control agents, or restraint of animals required by the other imaging
methods. The GFP imaging technology presented in this report will facilitate
studies of modulators of pancreatic cancer growth, including inhibition by
potential chemotherapeutic agents. -----------------------------------------------
The human pancreatic tumor cell line, BxPC-3, was engineered to stably express high-levels of the Aquorea victoria green fluorescent protein (GFP). The GFP-expressing pancreatic tumor cell line was surgically orthotopically implanted (SOI) as tissue fragments in the body of the pancreas of nude mice. Intravital imaging was used for quantification of growth metastasis on the liver. The diameters of the three micrometastases were 288 mm and 208 mm the right lobe and 344 mm on the left lobe as quantified by image analysis at day-70 post SOI. |
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Visualization of GFP-Expressing Tumors and Metastasis In Vivo
Robert M. Hoffman
[BIOTECHNIQUES 30, No. 5 May,
1016-1016, 2001]
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ABSTRACT: We have developed mouse
models of metastatic cancer with genetically fluorescent tumors that can be
imaged in fresh tissue, in situ, as
well as externally. To achieve this
capability, we have transduced the green fluorescent protein (GFP) gene,
cloned from the bioluminescent jellyfish Aequorea
victoria, into a series of human and rodent cancer cell lines that were
selected in vitro to stably express
GFP in vivo after transplantation
to metastatic rodent models. Techniques were also developed for transduction
of tumors by GFP in vivo. With this fluorescent tool, we detected and
visualized for the first time tumors and metastasis in fresh viable tissue or
in situ in host organs down to the single cell level. GFP tumors on the colon, prostate, breast,
brain, liver, lymph nodes, lung, pancreas, bone and other organs can also be
visualized externally, transcutaneously by quantitative whole-body
fluorescence optical imaging.
Real-time tumor and metastatic growth and angiogenesis and inhibition
by representative drugs can be imaged and quantified for rapid anti-tumor,
anti-metastatic and anti-angiogenesis drug screening. The GFP-transfected tumor cells enabled a
fundamental advance in the visualization of tumor growth and metastasis in
real time in vivo. -------------------------------------------
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Three-dimensional histoculture: Origins and applications in cancer research. Cancer Cells 3, 86-92, 1991.
Robert M. Hoffman
Cancer Cells 3, 86-92, 1991.
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Cover: Confocal photomicrograph of human colon cancer in three-dimensional histoculture on collagen containing sponges. The culture on the right has been treated with 5-fluorouracil. Viable cells are stained with BCEF-AM (green) and nonviable cells are stained with propidium iodide (red). For a review of methods and applications of three-dimensional histoculture, see the article.
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