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Systemic delivery of protein nanocages bearing CTT peptides for enhanced imaging of MMP-2 expression in metastatic tumor models.

Kawano T, Murata M, Piao JS, Narahara S, Hamano N, Kang JH, Hashizume M - Int J Mol Sci (2014)

Bottom Line: Here, we describe a metastatic cancer cell-targeted protein nanocage.An MMP-2-binding peptide, termed CTT peptide (CTTHWGFTLC), was conjugated to the surface of a naturally occurring heat shock protein nanocage by genetic modification.The engineered protein nanocages showed a binding affinity for MMP-2 and selective uptake in cancer cells that highly expressed MMP-2 in vitro.

View Article: PubMed Central - PubMed

Affiliation: Innovation Center for Medical Redox Navigation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. t-kawano@dem.med.kyushu-u.ac.jp.

ABSTRACT
Matrix metalloproteinase 2 (MMP-2) in metastatic cancer tissue, which is associated with a poor prognosis, is a potential target for tumor imaging in vivo. Here, we describe a metastatic cancer cell-targeted protein nanocage. An MMP-2-binding peptide, termed CTT peptide (CTTHWGFTLC), was conjugated to the surface of a naturally occurring heat shock protein nanocage by genetic modification. The engineered protein nanocages showed a binding affinity for MMP-2 and selective uptake in cancer cells that highly expressed MMP-2 in vitro. In near-infrared fluorescence imaging, the nanocages showed specific and significant accumulation in tumor tissue after intravenous injection in vivo. These protein nanocages conjugated with CTT peptide could be potentially applied to a noninvasive near-infrared fluorescence detection method for imaging gelatinase activity in metastatic tumors in vivo.

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Biodistribution of the protein nanocages in vivo. (a) In vivo real-time NIR fluorescence imaging of intravenously injected Alexa Fluor 750-labeled HspG41C-CTT in HT1080 and HT29 tumor-bearing mice. Time-dependent, tumor-targeting specificities of the nanocages were monitored by the IVIS system. Square regions indicate solid tumor growths of subcutaneously injected cancer cells; (b) Fluorescence signal intensity ratio of the tumor/background obtained from in vivo images. * p < 0.05; ** p < 0.01; (c) Organ sections of tumor-bearing mice injected intravenously with Alexa Fluor 488-labeled HspG41C-CTT at 3 h post-injection.
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ijms-16-00148-f005: Biodistribution of the protein nanocages in vivo. (a) In vivo real-time NIR fluorescence imaging of intravenously injected Alexa Fluor 750-labeled HspG41C-CTT in HT1080 and HT29 tumor-bearing mice. Time-dependent, tumor-targeting specificities of the nanocages were monitored by the IVIS system. Square regions indicate solid tumor growths of subcutaneously injected cancer cells; (b) Fluorescence signal intensity ratio of the tumor/background obtained from in vivo images. * p < 0.05; ** p < 0.01; (c) Organ sections of tumor-bearing mice injected intravenously with Alexa Fluor 488-labeled HspG41C-CTT at 3 h post-injection.

Mentions: Targeting tumor cells or tumor vasculature by peptides is a promising strategy to deliver therapeutic drugs and imaging agents for cancer therapy. CTT peptide-modified protein nanocages showed specific binding to cancer cells that expressed high levels of MMP-2 in vitro. To obtain in vivo evidences of tumor targeting, we examined the in vivo biodistribution of Alexa Fluor 750-labeled HspG41C-CTT nanocages in tumor-bearing mice. HT1080 and HT29 cells were used as positive and negative controls for MMP-2 expression, respectively. The NIR light is absorbed minimally by intrinsic chromophores, such as hemoglobin (<650 nm) and water (>900 nm), and thus result in noninvasive live animal imaging. Fluorescence imaging using NIR fluorescent agents displays relatively low autofluorescence. Immediately after intravenous injection, HspG41C-CTT showed continuous accumulation in tumors, and NIR fluorescence intensities reached a maximum at 3 h post-injection (Figure 5a,b). However, in HT29 tumor-bearing mice as the negative control, HspG41C-CTT showed less accumulation in tumors. Examination of tissue sections obtained at 3 h post-injection revealed HspG41C-CTT nanocages in HT1080 tumor tissues (Figure 5c), whereas non-targeted HspG41C nanocages had accumulated in both types of tumors (data not shown). The distributions of HspG41C after intravenous injection were similar to a previous report of accumulation in the liver and rapid excretion through kidney [28].


Systemic delivery of protein nanocages bearing CTT peptides for enhanced imaging of MMP-2 expression in metastatic tumor models.

Kawano T, Murata M, Piao JS, Narahara S, Hamano N, Kang JH, Hashizume M - Int J Mol Sci (2014)

Biodistribution of the protein nanocages in vivo. (a) In vivo real-time NIR fluorescence imaging of intravenously injected Alexa Fluor 750-labeled HspG41C-CTT in HT1080 and HT29 tumor-bearing mice. Time-dependent, tumor-targeting specificities of the nanocages were monitored by the IVIS system. Square regions indicate solid tumor growths of subcutaneously injected cancer cells; (b) Fluorescence signal intensity ratio of the tumor/background obtained from in vivo images. * p < 0.05; ** p < 0.01; (c) Organ sections of tumor-bearing mice injected intravenously with Alexa Fluor 488-labeled HspG41C-CTT at 3 h post-injection.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4307240&req=5

ijms-16-00148-f005: Biodistribution of the protein nanocages in vivo. (a) In vivo real-time NIR fluorescence imaging of intravenously injected Alexa Fluor 750-labeled HspG41C-CTT in HT1080 and HT29 tumor-bearing mice. Time-dependent, tumor-targeting specificities of the nanocages were monitored by the IVIS system. Square regions indicate solid tumor growths of subcutaneously injected cancer cells; (b) Fluorescence signal intensity ratio of the tumor/background obtained from in vivo images. * p < 0.05; ** p < 0.01; (c) Organ sections of tumor-bearing mice injected intravenously with Alexa Fluor 488-labeled HspG41C-CTT at 3 h post-injection.
Mentions: Targeting tumor cells or tumor vasculature by peptides is a promising strategy to deliver therapeutic drugs and imaging agents for cancer therapy. CTT peptide-modified protein nanocages showed specific binding to cancer cells that expressed high levels of MMP-2 in vitro. To obtain in vivo evidences of tumor targeting, we examined the in vivo biodistribution of Alexa Fluor 750-labeled HspG41C-CTT nanocages in tumor-bearing mice. HT1080 and HT29 cells were used as positive and negative controls for MMP-2 expression, respectively. The NIR light is absorbed minimally by intrinsic chromophores, such as hemoglobin (<650 nm) and water (>900 nm), and thus result in noninvasive live animal imaging. Fluorescence imaging using NIR fluorescent agents displays relatively low autofluorescence. Immediately after intravenous injection, HspG41C-CTT showed continuous accumulation in tumors, and NIR fluorescence intensities reached a maximum at 3 h post-injection (Figure 5a,b). However, in HT29 tumor-bearing mice as the negative control, HspG41C-CTT showed less accumulation in tumors. Examination of tissue sections obtained at 3 h post-injection revealed HspG41C-CTT nanocages in HT1080 tumor tissues (Figure 5c), whereas non-targeted HspG41C nanocages had accumulated in both types of tumors (data not shown). The distributions of HspG41C after intravenous injection were similar to a previous report of accumulation in the liver and rapid excretion through kidney [28].

Bottom Line: Here, we describe a metastatic cancer cell-targeted protein nanocage.An MMP-2-binding peptide, termed CTT peptide (CTTHWGFTLC), was conjugated to the surface of a naturally occurring heat shock protein nanocage by genetic modification.The engineered protein nanocages showed a binding affinity for MMP-2 and selective uptake in cancer cells that highly expressed MMP-2 in vitro.

View Article: PubMed Central - PubMed

Affiliation: Innovation Center for Medical Redox Navigation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. t-kawano@dem.med.kyushu-u.ac.jp.

ABSTRACT
Matrix metalloproteinase 2 (MMP-2) in metastatic cancer tissue, which is associated with a poor prognosis, is a potential target for tumor imaging in vivo. Here, we describe a metastatic cancer cell-targeted protein nanocage. An MMP-2-binding peptide, termed CTT peptide (CTTHWGFTLC), was conjugated to the surface of a naturally occurring heat shock protein nanocage by genetic modification. The engineered protein nanocages showed a binding affinity for MMP-2 and selective uptake in cancer cells that highly expressed MMP-2 in vitro. In near-infrared fluorescence imaging, the nanocages showed specific and significant accumulation in tumor tissue after intravenous injection in vivo. These protein nanocages conjugated with CTT peptide could be potentially applied to a noninvasive near-infrared fluorescence detection method for imaging gelatinase activity in metastatic tumors in vivo.

Show MeSH
Related in: MedlinePlus