Intravital imaging of human prostate cancer using viral nanoparticles targeted to gastrin-releasing Peptide receptors.

Multivalent nanoparticles have several key advantages in terms of solubility, binding avidity, and uptake, making them particularly well suited to molecular imaging applications. Herein is reported the stepwise synthesis and characterization of NIR viral nanoparticles targeted to gastrin-releasing peptide receptors that are over-expressed in human prostate cancers. The pan-bombesin analogue, [β-Ala11, Phe13, Nle14]bombesin-(7-14), is conjugated to cowpea mosaic virus particles functionalized with an NIR dye (Alexa Fluor 647) and polyethylene glycol (PEG) using the copper(I)-catalyzed azide-alkyne cycloaddition reaction. Targeting and uptake in human PC-3 prostate cells is demonstrated in vitro. Tumor homing is observed using human prostate tumor xenografts on the chicken chorioallantoic membrane model using intravital imaging. Further development of this viral nanoparticle platform may open the door to potential clinical noninvasive molecular imaging strategies.

[1]  E. Krenning,et al.  Novel 111In-labelled bombesin analogues for molecular imaging of prostate tumours , 2007, European Journal of Nuclear Medicine and Molecular Imaging.

[2]  S. St.-Pierre,et al.  The contractile effect of bombesin, gastrin releasing peptide and various fragments in the rat stomach strip. , 1984, European journal of pharmacology.

[3]  Steve Y. Cho,et al.  New Agents and Techniques for Imaging Prostate Cancer , 2009, Journal of Nuclear Medicine.

[4]  N. Steinmetz,et al.  Structure-based engineering of an icosahedral virus for nanomedicine and nanotechnology. , 2009, Current topics in microbiology and immunology.

[5]  G. Ferro-Flores,et al.  Preparation and evaluation of 99mTc-EDDA/HYNIC-[Lys3]-bombesin for imaging gastrin-releasing peptide receptor-positive tumours , 2006, Nuclear medicine communications.

[6]  N. Steinmetz,et al.  Buckyballs meet viral nanoparticles: candidates for biomedicine. , 2009, Journal of the American Chemical Society.

[7]  R A Sayle,et al.  RASMOL: biomolecular graphics for all. , 1995, Trends in biochemical sciences.

[8]  M. Finn,et al.  Analysis and optimization of copper-catalyzed azide-alkyne cycloaddition for bioconjugation. , 2009, Angewandte Chemie.

[9]  N. Steinmetz,et al.  Chemical Introduction of Reactive Thiols Into a Viral Nanoscaffold: A Method that Avoids Virus Aggregation , 2007, Chembiochem : a European journal of chemical biology.

[10]  A. Beck‐Sickinger,et al.  In vitro and in vivo evaluation of a 99mTc(I)-labeled bombesin analogue for imaging of gastrin releasing peptide receptor-positive tumors. , 2002, Nuclear medicine and biology.

[11]  J. Lewis,et al.  The inhibition of tumor cell intravasation and subsequent metastasis via regulation of in vivo tumor cell motility by the tetraspanin CD151. , 2008, Cancer cell.

[12]  John E. Johnson,et al.  Virus Particle Explorer (VIPER), a Website for Virus Capsid Structures and Their Computational Analyses , 2001, Journal of Virology.

[13]  Q. Wang,et al.  A fluorogenic 1,3-dipolar cycloaddition reaction of 3-azidocoumarins and acetylenes. , 2004, Organic letters.

[14]  Q. Wang,et al.  Icosahedral Virus Particles as Polyvalent Carbohydrate Display Platforms , 2003, Chembiochem : a European journal of chemical biology.

[15]  R. Weissleder,et al.  Multivalent effects of RGD peptides obtained by nanoparticle display. , 2006, Journal of medicinal chemistry.

[16]  R. Roesler,et al.  Gastrin-releasing peptide receptor as a molecular target in experimental anticancer therapy. , 2007, Annals of oncology : official journal of the European Society for Medical Oncology.

[17]  G. Slegers,et al.  Technetium-99m RP527, a GRP analogue for visualisation of GRP receptor-expressing malignancies: a feasibility study , 2000, European Journal of Nuclear Medicine.

[18]  P. Singh,et al.  Virus-based nanoparticles (VNPs): platform technologies for diagnostic imaging. , 2006, Advanced drug delivery reviews.

[19]  J. Lewis,et al.  Synthesis of bombesin-functionalized iron oxide nanoparticles and their specific uptake in prostate cancer cells , 2010, Journal of nanoparticle research : an interdisciplinary forum for nanoscale science and technology.

[20]  Donald L. Hayes,et al.  Radiochemical investigations of gastrin-releasing peptide receptor-specific [(99m)Tc(X)(CO)3-Dpr-Ser-Ser-Ser-Gln-Trp-Ala-Val-Gly-His-Leu-Met-(NH2)] in PC-3, tumor-bearing, rodent models: syntheses, radiolabeling, and in vitro/in vivo studies where Dpr = 2,3-diaminopropionic acid and X = H2O or P(CH2 , 2003, Cancer research.

[21]  V. Erspamer,et al.  Isolation and structure of bombesin and alytesin, two analogous active peptides from the skin of the european amphibiansBombina andAlytes , 1971, Experientia.

[22]  H. Wagner,et al.  Design, synthesis, and initial evaluation of high-affinity technetium bombesin analogues. , 1998, Bioconjugate chemistry.

[23]  John E. Johnson,et al.  New addresses on an addressable virus nanoblock; uniquely reactive Lys residues on cowpea mosaic virus. , 2004, Chemistry & biology.

[24]  H. Maeda,et al.  Exploiting the enhanced permeability and retention effect for tumor targeting. , 2006, Drug discovery today.

[25]  A. Jemal,et al.  Cancer Statistics, 2008 , 2008, CA: a cancer journal for clinicians.

[26]  N. Steinmetz,et al.  Hydrazone ligation strategy to assemble multifunctional viral nanoparticles for cell imaging and tumor targeting. , 2010, Nano letters.

[27]  J. Reubi,et al.  Bombesin Receptor Subtypes in Human Cancers: Detection with the Universal Radioligand 125I-[d-TYR6, β-ALA11, PHE13, NLE14] Bombesin(6–14) , 2002 .

[28]  Marianne Manchester,et al.  Folic acid-mediated targeting of cowpea mosaic virus particles to tumor cells. , 2007, Chemistry & biology.

[29]  G. Erspamer,et al.  RELATIVE POTENCY OF BOMBESIN‐LIKE PEPTIDES , 1975, British journal of pharmacology.

[30]  Gary Siuzdak,et al.  Endothelial Targeting of Cowpea Mosaic Virus (CPMV) via Surface Vimentin , 2009, PLoS pathogens.

[31]  N. Steinmetz,et al.  PEGylated viral nanoparticles for biomedicine: the impact of PEG chain length on VNP cell interactions in vitro and ex vivo. , 2009, Biomacromolecules.

[32]  Andries Zijlstra,et al.  Viral nanoparticles as tools for intravital vascular imaging , 2006, Nature Medicine.

[33]  S. Chevalier,et al.  Bombesin specifically induces intracellular calcium mobilization via gastrin-releasing peptide receptors in human prostate cancer cells. , 1996, Journal of molecular endocrinology.

[34]  N. Steinmetz,et al.  Cowpea mosaic virus nanoparticles target surface vimentin on cancer cells. , 2011, Nanomedicine.

[35]  H. Maeda,et al.  Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[36]  M. Finn,et al.  Virus-glycopolymer conjugates by copper(I) catalysis of atom transfer radical polymerization and azide-alkyne cycloaddition. , 2005, Chemical communications.

[37]  Kristopher J. Koudelka,et al.  Interaction between a 54-Kilodalton Mammalian Cell Surface Protein and Cowpea Mosaic Virus , 2006, Journal of Virology.

[38]  S. Mather,et al.  Targeting prostate cancer with radiolabelled bombesins , 2006, Cancer imaging : the official publication of the International Cancer Imaging Society.