Near-Infrared Optical Imaging of Integrin αvβ3 in Human Tumor Xenografts

In vivo optical imaging is potentially useful for evaluating the presence of tumor markers that are targets of molecular medicine. Here we report the synthesis and characterization of integrin alphavbeta3-targeted peptide cyclo(Lys-Arg-Gly-Asp-Phe) [c(KRGDf )] labeled with fluorescence dyes with wavelength spanning from the visible/near infrared (Cy5.5) to the true near infrared (IRDye800) for optical imaging. In vitro, the peptide-dye conjugates bound specifically to tumor cells expressing alphavbeta3. When administered intravenously into mice at a dose of 6 nmol /mouse, the conjugates accumulated in tumors expressing alphavbeta3. The tumor-to-background ratios for human KS1767 Kaposi's sarcoma in mice injected with Cy5.5-c(KRGDf ) and Cy5.5 were 5.5 and 1.5, respectively. Preinjection of c(KRGDf ) blocked the uptake of Cy5.5-c(KRGDf ) in tumors by 89%. In alphavbeta3-positive M21 and alphavbeta3-negative M21-L human melanoma, fluorescence intensity in the tumor of mice injected with IRDye800 - c(KRGDf ) was 2.3 and 1.3 times that in normal tissue, respectively. Dynamic imaging revealed that Cy5.5- c(KRGDf ) was rapidly taken up by KS1767 tumor immediately after bolus injection. The rate of its uptake in the tumor was reduced by preinjection of c(KRGDf ) in an interval time-dependent manner. Our data suggest that near-infrared fluorescence imaging may be applied to the detection of tumors expressing integrin alphavbeta3 and to the assessment of the optimal biological dose and schedule of targeted therapies.

[1]  A. Shilkaitis,et al.  Beta3 integrin expression in melanoma predicts subsequent metastasis. , 1996, The Journal of surgical research.

[2]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[3]  David A. Cheresh,et al.  Detection of tumor angiogenesis in vivo by αvβ3-targeted magnetic resonance imaging , 1998, Nature Medicine.

[4]  M. Schwaiger,et al.  Synthesis and biological evaluation of a 99mTc-labelled cyclic RGD peptide for imaging the αvβ3 expression , 2004, Nuklearmedizin.

[5]  A B Thompson,et al.  Diagnostic imaging of breast cancer using fluorescence-enhanced optical tomography: phantom studies. , 2004, Journal of biomedical optics.

[6]  D. Cheresh.  Human endothelial cells synthesize and express an Arg-Gly-Asp-directed adhesion receptor involved in attachment to fibrinogen and von Willebrand factor. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Brugge,et al.  Integrins and signal transduction pathways: the road taken. , 1995, Science.

[8]  R. Weissleder,et al.  In vivo imaging of tumors with protease-activated near-infrared fluorescent probes , 1999, Nature Biotechnology.

[9]  Ryan Park,et al.  MicroPET and autoradiographic imaging of breast cancer alpha v-integrin expression using 18F- and 64Cu-labeled RGD peptide. , 2004, Bioconjugate chemistry.

[10]  A. Halpern,et al.  A Prognostic Model for Predicting 10-Year Survival in Patients with Primary Melanoma , 1996, Annals of Internal Medicine.

[11]  E. Sevick-Muraca,et al.  Near-Infrared Fluorescence Optical Imaging and Tomography , 2004, Disease markers.

[12]  Daniel L. Farkas,et al.  Tumor labeling in vivo using cyanine-conjugated monoclonal antibodies , 1995, Cancer Immunology, Immunotherapy.

[13]  R. Taillefer,et al.  Imaging characteristics of a novel technetium Tc 99m-labeled platelet glycoprotein IIb/IIIa receptor antagonist in patients With acute deep vein thrombosis or a history of deep vein thrombosis. , 2003, Archives of internal medicine.

[14]  Chun Li,et al.  Near-infrared optical imaging of epidermal growth factor receptor in breast cancer xenografts. , 2003, Cancer research.

[15]  D. Cheresh,et al.  Involvement of integrin alpha V gene expression in human melanoma tumorigenicity. , 1992, The Journal of clinical investigation.

[16]  D. Cheresh,et al.  The role of alphav integrins during angiogenesis: insights into potential mechanisms of action and clinical development. , 1999, The Journal of clinical investigation.

[17]  David A. Cheresh,et al.  Definition of Two Angiogenic Pathways by Distinct αv Integrins , 1995, Science.

[18]  Christoph Ballestrem,et al.  Marching at the front and dragging behind , 2001, The Journal of cell biology.

[19]  Robert E. Lenkinski,et al.  In vivo near-infrared fluorescence imaging of osteoblastic activity , 2001, Nature Biotechnology.

[20]  Horst Kessler,et al.  Radiolabeled αvβ3 Integrin Antagonists: A New Class of Tracers for Tumor Targeting , 1999 .

[21]  D. Ruiter,et al.  Establishment and characterization of a human melanoma cell line (MV3) which is highly metastatic in nude mice , 2007, International journal of cancer.

[22]  M Schwaiger,et al.  Glycosylated RGD-containing peptides: tracer for tumor targeting and angiogenesis imaging with improved biokinetics. , 2001, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[23]  J. Bading,et al.  Pharmacokinetics and tumor retention of 125I-labeled RGD peptide are improved by PEGylation. , 2004, Nuclear medicine and biology.

[24]  E Ruoslahti,et al.  New perspectives in cell adhesion: RGD and integrins. , 1987, Science.

[25]  J. Schneider-Mergener,et al.  Cyanine Dye Labeled Vasoactive Intestinal Peptide and Somatostatin Analog for Optical Detection of Gastroenteropancreatic Tumors , 2000, Annals of the New York Academy of Sciences.

[26]  D. Cheresh,et al.  Requirement of vascular integrin alpha v beta 3 for angiogenesis. , 1994, Science.

[27]  F. Sarkar,et al.  Antiintegrin alpha v beta 3 blocks human breast cancer growth and angiogenesis in human skin. , 1995, The Journal of clinical investigation.

[28]  Milind Rajopadhye,et al.  Tumor targeting with radiolabeled alpha(v)beta(3) integrin binding peptides in a nude mouse model. , 2002, Cancer research.

[29]  M. Schwaiger,et al.  [18F]Galacto-RGD: synthesis, radiolabeling, metabolic stability, and radiation dose estimates. , 2004, Bioconjugate chemistry.

[30]  R. Weissleder,et al.  Charge-coupled-device based scanner for tomography of fluorescent near-infrared probes in turbid media. , 2002, Medical physics.

[31]  S. Achilefu,et al.  Novel receptor-targeted fluorescent contrast agents for in vivo tumor imaging. , 2000, Investigative radiology.

[32]  Sibylle Ziegler,et al.  Noninvasive Imaging of αvβ3 Integrin Expression Using 18F-labeled RGD-containing Glycopeptide and Positron Emission Tomography , 2001 .

[33]  H. Bergh,et al.  Antibody-indocyanin conjugates for immunophotodetection of human squamous cell carcinoma in nude mice. , 1994, Cancer research.