Oncology
Courtesy of Frauke Alves
Department of Hematology and Oncology, University Medical Center
Göttingen, Germany
Monitoring tumor growth using antibody-based fluorescence imaging and fpVCT
Representative fluorescence images and flat-panel volume computed tomography (fpVCT) scans of a nude mouse were obtained weekly after orthotopic implantation of human MDA-MB-231 mammary carcinoma cells. This study investigates if combining imaging techniques can allow for the visualization of functional information obtained by fluorescence imaging within an adequate anatomic framework.
Top row: Normalized fluorescence intensity measurements obtained 3, 4, and 5 weeks post tumor implantation and 24 hours after injection of uPAR-mAb*Cy5.5, a Cy5.5 labeled tumor-specific anti uPAR antibody.
Bottom row: Contrast-enhanced fpVCT scans obtained on the same days.
Paper
Concept of selective tumour therapy and its evaluation by near-infrared fluorescence imaging and flat-panel volume computed tomography in mice
Frauke Alves, Christian Dullin, Joanna Napp, Jeannine Missbach-Guentner, Katharina Jannasch, Julia Mathejczyk, Luis A. Pardo, Walter Stühmer, and Lutz-F. Tietze. European Journal of Radiology, Volume 70, Number 2 / May 2009: pp. 286-293.
Courtesy of Alain Le Pape
Centre national de la recherche scientifique (National Center for Scientific Research)
Orleans, France
Pulmonary distribution of nebulized Cetuximab labeled with Xenofluor 680
Cetuximab, a chimeric monoclonal antibody that can be used to target EGFR in presence of tumors cells, was used to test nebulization as a means to deliver therapy through the airways for lung cancer treatment. Two mice were subjected to nebulization with Cetuximab labeled with Xenofluor (90 μg/200 μl) and imaged at different times. The in vivo assay performed shows that the distribution of Cetuximab in the lungs was not uniform.
Left: Intensity image map of nebulized mouse. Note that intensity is observed in the right lung only.
Right: Lifetime image shows Cetuximab is in fact in both lungs.
Courtesy of Alain Le Pape
Centre national de la recherche scientifique (National Center for Scientific Research)
Orleans, France
3D concentration distribution of nebulized Cetuximab in mouse lungs
Cetuximab, a chimeric monoclonal antibody that can be used to target EGFR in presence of tumors cells, was used to test nebulization as a means to deliver therapy through the airways for lung cancer treatment. Two mice were subjected to nebulization with Cetuximab labeled with Xenofluor (90 μg/200 μl) and imaged at different times. The in vivo assay performed shows that the distribution of Cetuximab in the lungs was not uniform.
Image: Relative quantification of the Cetuximab-Xenofluor 680 concentration volume detected in each lung using the OptiView concentration analysis tool. The goal of the study was to evaluate optical imaging as an alternative to microPet for quantification measurements.
Courtesy of Joanna Napp
University of Göttingen
Germany
Monitoring proteolytic enzymes as a novel target for anticancer therapy
In this study, 1x106 human pancreatic cancer AsPC-1 cells expressing matriptase were orthotopically implanted into male nude mice. Prior to scanning, either 25 mg of polyclonal Cy5.5 labeled anti-human matriptase antibody (MAD*Cy5.5) or 4 nmol of the fluorescent substrate (S*DY-681) were injected intravenously. Binding of the antibody and thereby matriptase expression in vivo was determined by measurement of fluorescence intensity, lifetime, and location in the tumor-bearing mice over time and confirmed by histological analysis.
Images: Binding kinetics of MAD*Cy5.5 to AsPC-1 cells in vivo. Fluorescence intensity (A) at various time points after MAD*Cy5.5 injection. To exclude unspecific signals, the fluorescence lifetime was gated between 1.6 and 2.0 ns. Correlation of the signal (B) with macroscopic appearance of the primary pancreatic tumor (circled).
Poster
Evaluation of Novel Targeted Tumor Therapies by Non-Invasive Near Infrared Imaging
Joanna Napp, Christian Dullin, Sarah Kimmina, Friedemann Müller, Andres van de Locht, Torsten Steinmetzer, and Frauke Alves. European Society for Molecular Imaging, Naples, Italy, June 2007.
Courtesy of Joanna Napp
University of Göttingen
Germany
Monitoring proteolytic enzymes as a novel target for anticancer therapy
In this study, 1x106 human pancreatic cancer AsPC-1 cells expressing matriptase were orthotopically implanted into male nude mice. Prior to scanning, either 25 mg of polyclonal Cy5.5 labeled anti-human matriptase antibody (MAD*Cy5.5) or 4 nmol of the fluorescent substrate (S*DY-681) were injected intravenously. Binding of the antibody and thereby matriptase expression in vivo was determined by measurement of fluorescence intensity, lifetime, and location in the tumor-bearing mice over time and confirmed by histological analysis.
Images: Correlation of (D) in vivo fluorescent signals to (A) the pancreatic tumor (circled in white) and metastatic sites in the liver (red) and bladder (black) or scar (C, D).
Poster
Evaluation of Novel Targeted Tumor Therapies by Non-Invasive Near Infrared Imaging
Joanna Napp, Christian Dullin, Sarah Kimmina, Friedemann Müller, Andres van de Locht, Torsten Steinmetzer, and Frauke Alves. European Society for Molecular Imaging, Naples, Italy, June 2007.
Courtesy of In-Sam Kim
Department of Biochemistry and Cell Biology, Kyungpook National University School of Medicine
Daegu, Korea
In vivo imaging of drug-induced apoptosis using a PS-recognizing peptide
Tumor-bearing mice were either left untreated or treated with a single dose of the apoptosis inducing chemical camptothecin (10 mg/kg) 24 hours prior to intravenous injections of a fluorescein-labeled PS-recognizing peptide (CLSYYPSYC) or control peptides. For each mouse, in vivo baseline fluorescence was assessed prior to peptide injection, and at 2, 6, and 18 hours post-injection.
Top row: Fluorescence intensities of treated mouse injected with CLSTTPSYC.
Middle row: Fluorescence intensities of untreated mouse injected with CLSTTPSYC.
Bottom row: Fluorescence intensities of treated mouse injected with a control peptide.
Application Note
ART's Optix Small Animal Imager Applied to Non-Invasive Apoptosis Research in Mouse Xenograft Model (PDF: 707 Kb)
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Paper
Discovery of a Phosphatidylserine Recognizing Peptide and Its Utility in Molecular Imaging of Tumor Apoptosis
Narendra Thapa, Soyoun Kim, In-Sup So, Byung-Heon Lee, Ick-Chan Kwon, Kuiwon Choi, and In-San Kim. Journal of Cellular and Molecular Medicine, Volume 12, Number 5A / September-October 2008: pp. 1649-1660.
ART Advanced Research Technologies Inc.
Quantitative analysis of multiple tumor-targeting agents using fluorescence lifetime
A U87MG glioblastoma cell line with a mutation in the EGF receptor was xenografted to nude mice and allowed to grow for 21 days. The mice were then injected intravenously through the tail vein with a contrast agent conjugated with either a C225 antibody (Cetuximab) or a transferrin (Tf) protein. A third category of mice received an injection of C225-Cy5.5, followed by a Tf-DY682 injection 44 hours later. To boost the signal of Tf-DY682, a second dose was injected 24 hours later.
Images: Fluorescence intensities of C225-Cy5.5 in liver (left) and in tumor (middle), and of Tf-DY682 in tumor (right).
Application Note
ART Optix quantitative analysis of multiple tumor targeting agents using fluorescence lifetime (PDF: 459 Kb)
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ART Advanced Research Technologies Inc.
Quantitative analysis of multiple tumor-targeting agents using fluorescence lifetime
A U87MG glioblastoma cell line with a mutation in the EGF receptor was xenografted to nude mice and allowed to grow for 21 days. The mice were then injected intravenously through the tail vein with a contrast agent conjugated with either a C225 antibody (Cetuximab) or a transferrin (Tf) protein. A third category of mice received an injection of C225-Cy5.5, followed by a Tf-DY682 injection 44 hours later. To boost the signal of Tf-DY682, a second dose was injected 24 hours later.
Images: Fluorescence lifetime (ns) image maps of C225-Cy5.5 in the liver (left) and tumor (middle), and of Tf-DY682 in tumor (right). Note that the unbound deactivated Cy5.5 dye has a lifetime value of about 1.6 ns, while that of the conjugated C225-Cy5.5 probe is approximately 2.0 ns. The lifetime of Tf-tagged DY682 is around 2.6 ns. Although a fluorescent signal is detected in both the liver and tumor, lifetime analysis determined that the bound probe is found only in the tumor region and that the signal localized to the liver shortly after the injection can be attributed to metabolized free dye. Similarly, the signals of different dyes in the tumor region can also be differentiated by their lifetimes. By applying lifetime gating, which can be used to selectively display signals with specific lifetime values, the signals from free Cy5.5, C225-conjugated Cy5.5, and Tf-tagged DY682 are clearly distinguishable.
Application Note
ART Optix quantitative analysis of multiple tumor targeting agents using fluorescence lifetime (PDF: 459 Kb)
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Courtesy of Abedelnasser Abulrob
National Research Council of Canada, Institute for Biological Sciences
Ottawa, Canada
Imaging of brain tumors using anti-EGFR single domain antibodies
Left: U87MG human glioblastoma cells over-expressing EGFR or EGFRvIII were injected intracranially into the right striatum of nude CD-1 mice. After the tumor was developed for 2 weeks, Cy5.5 labeled 225 Mab and V2C sdAb were injected by tail vein and imaged at various time points. Fluorescence image obtained 24 hours after injection of 225 Mab.
Right: U87MG human glioblastoma cells over-expressing EGFR or EGFRvIII were injected subcutaneously into the left flank of nude CD-1 mice. After the tumor was developed for 10 days, Cy5.5 labeled anti-EGFR V2C sdAb variants were injected by tail vein and imaged at various time points. Fluorescence image obtained 24 hours after injection of V2C pentameric sdAb.
Poster
Molecular Imaging of Brain Tumors Using Single-Domain Antibodies
Umar Iqbal, Abedelnasser Abulrob, Ulrike Trojahn, Jianbing Zhang, Maureen O’Connor-McCourt, Roger Mackenzie, and Danica Stanimirovic. Joint Conference of the Academy of Molecular Imaging and the Society of Molecular Imaging, Providence, Rhode Island, USA, Sept 2007.
ART Advanced Research Technologies Inc.
Assessing murine lung tumor burden with fluorescent mAb therapy
This experiment involved three animals (M1, M2, and M3) that were induced with tumors via a tail vein injection of the human xenograft cell line NIH-H11650, one negative control (M185) that had no tumor burden, and a transgenic mouse (M6098) of unknown tumor status. Cetuximab, an EGFR inhibitor, conjugated to Alexa Fluor 680 was used as the reagent for tumor localization. The distribution and localization over various time points was determined after Cetuximab-AF680 injection.
Top row: Comparison of normalized intensity images pre-injection (prescan) and 2 hrs post-injection for animals M1, M2, and M6098.
Bottom row: Normalized intensity images lifetime gated between 1.32 and 1.46 ns for the same images as above. By gating out the fluorescence signal from other sources, the signal of Cetuximab-AF680 localizing to the tumor was isolated and observed.
Application Note
Assessing murine lung tumor burden with fluorescent mAb therapy using the ART Optix, an in vivo fluorescent time-domain imaging system (PDF: 931 Kb)
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ART Advanced Research Technologies Inc.
Fluorescence lifetime assessment of Cetuximab-Alexa Fluor 680
This experiment involved three animals (M1, M2, and M3) that were induced with tumors via a tail vein injection of the human xenograft cell line NIH-H11650, one negative control (M185) that had no tumor burden, and a transgenic mouse (M6098) of unknown tumor status. Cetuximab, an EGFR inhibitor, conjugated to Alexa Fluor 680 was used as the reagent for tumor localization. The distribution and localization over various time points was followed after Cetuximab-AF680 injection.
Images: Normalized intensity images lifetime gated between 1.32 and 1.46 ns comparing M1 and M185 over time. Cetuximab-AF680 shows clear specificity for tumor and does not bind to lung tissue non-specifically in the absence of tumor.
Application Note
Assessing murine lung tumor burden with fluorescent mAb therapy using the ART Optix, an in vivo fluorescent time-domain imaging system (PDF: 931 Kb)
Log in to download ART application notes.
“Our results show that fluorescence lifetime imaging (FLI) can be useful for the dynamic imaging of protein-losing nephropathy due to diabetes mellitus and other renal diseases and suggest potential use of FLI to distinguish tumors from fluid-filled cysts in the body.”
“The lifetime of rituximab-Cy5.5 is a parameter sufficiently sensitive to discriminate between the exogenous signal of rituximab-Cy5.5 and auto-fluorescence and to monitor changes in the tumor tissue microenvironment”
Be Quantitative
From oncology and neurological disease models to pharmacokinetics, the Optix MX3 system captures more data than any other in vivo imaging system.