The biodistribution of self-assembling protein nanoparticles shows they are promising vaccine platforms.
BACKGROUND
Because of the need to limit side-effects, nanoparticles are increasingly being studied as drug-carrying and targeting tools. We have previously reported on a scheme to produce protein-based self-assembling nanoparticles that can act as antigen display platforms. Here we attempted to use the same system for cancer-targeting, making use of a C-terminal bombesin peptide that has high affinity for a receptor known to be overexpressed in certain tumors, as well as an N-terminal polyhistidine tag that can be used for radiolabeling with technetium tricarbonyl.


RESULTS
In order to increase circulation time, we experimented with PEGylated and unPEGylated varities typo particle. We also tested the effect of incorporating different numbers of bombesins per nanoparticle. Biophysical characterization determined that all configurations assemble into regular particles with relatively monodisperse size distributions, having peaks of about 33-36 nm. The carbonyl method used for labeling produced approximately 80% labeled nanoparticles. In vitro, the nanoparticles showed high binding, both specific and non-specific, to PC-3 prostate cancer cells. In vivo, high uptake was observed for all nanoparticle types in the spleens of CD-1 nu/nu mice, decreasing significantly over the course of 24 hours. High uptake was also observed in the liver, while only low uptake was seen in both the pancreas and a tumor xenograft.


CONCLUSIONS
The data suggest that the nanoparticles are non-specifically taken up by the reticuloendothelial system. Low uptake in the pancreas and tumor indicate that there is little or no specific targeting. PEGylation or increasing the amount of bombesins per nanoparticle did not significantly improve targeting. In particular, the uptake in the spleen, which is a primary organ of the immune system, highlights the potential of the nanoparticles as vaccine carriers. Also, the decrease in liver and spleen radioactivity with time implies that the nanoparticles are broken down and cleared. This is an important finding, as it shows that the nanoparticles can be safely used as a vaccine platform without the risk of prolonged side effects. Furthermore, it demonstrates that technetium carbonyl radiolabeling of our protein-based nanoparticles can be used to evaluate their pharmacokinetic properties in vivo.
Yongkun
Y
Yang
Tobias
T
Neef
Christian
C
Mittelholzer
Elisa
E
Garcia Garayoa
Peter
P
Bläuenstein
Roger
R
Schibli
Ueli
U
Aebi
Department of Molecular and Cell Biology and Institute of Materials Science, University of Connecticut, 97 N, Eagleville Road, Storrs, CT 06250, USA. peter.burkhard@uconn.edu.
Peter
P
Burkhard
D000230:Adenocarcinoma / Q000276:immunology / Q000517:prevention & control; D000595:Amino Acid Sequence; D000818:Animals; D001839:Bombesin / Q000737:chemistry*; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000276:immunology; D016503:Drug Delivery Systems / Q000379:methods*; D005260:Female; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism; D010316:Particle Size; D010455:Peptides / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D011092:Polyethylene Glycols / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000276:immunology / Q000517:prevention & control; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D013154:Spleen / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D013194:Staining and Labeling; D013667:Technetium; D023041:Xenograft Model Antitumor Assays
D019496:Cancer Vaccines; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011092:Polyethylene Glycols; D013667:Technetium; D001839:Bombesin