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Enzyme-Controlled Intracellular Self-Assembly of (18)F Nanoparticles for Enhanced MicroPET Imaging of Tumor.

Liu Y, Miao Q, Zou P, Liu L, Wang X, An L, Zhang X, Qian X, Luo S, Liang G - Theranostics (2015)

Bottom Line: TEM images of 1-Cold-treated MDA-MB-468 cells directly uncovered that the intracellular 1-Cold-NPs were at/near the location of furin (i.e., Golgi bodies).MTT results indicated that 50 µM 1-Cold did not impose cytotoxicity to MDA-MB-468 cells up to 12 hours.Our ''smart'' probe (i.e., 1), together with the strategy of co-injection, might help researchers trace the biomarkers of interest within a longer time window.

View Article: PubMed Central - PubMed

Affiliation: 1. School of Chemical and Material Engineering, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China ; 3. Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.

ABSTRACT
Herein, we report the development of a new "smart" radioactive probe (i.e., 1) which can undergo furin-controlled condensation and self-assembly of radioactive nanoparticles (i.e., 1-NPs) in tumor cells and its application for enhanced microPET imaging of tumors in nude mice co-injected with its cold analog (i.e., 1-Cold). Furin-controlled condensation of 1-Cold and self-assembly of its nanoparticles (i.e., 1-Cold-NPs) in vitro were validated and characterized with HPLC, mass spectra, SEM, and TEM analyses. Cell uptake studies showed that both 1 and 1-Cold have good cell permeability. TEM images of 1-Cold-treated MDA-MB-468 cells directly uncovered that the intracellular 1-Cold-NPs were at/near the location of furin (i.e., Golgi bodies). MTT results indicated that 50 µM 1-Cold did not impose cytotoxicity to MDA-MB-468 cells up to 12 hours. MicroPET imaging of MDA-MB-468 tumor-bearing mice indicated that mice co-injected with 1 and 1-Cold showed higher uptake and longer attenuation of the radioactivity in tumors than those mice only injected with same dosage of 1. Tumor uptake ratios of 1 between these two groups of mice reached the maximum of 8.2 folds at 240 min post injection. Biodistribution study indicated that the uptake ratios of 1 in kidneys between these two groups continuously increased and reached 81.9 folds at 240 min post injection, suggesting the formation of radioactive NPs (i.e., 1-NPs) in MDA-MB-468 tumors of mice co-injected with 1 and 1-Cold. And the nanoparticles were slowly digested and secreted from the tumors, accumulating in the kidneys. Our ''smart'' probe (i.e., 1), together with the strategy of co-injection, might help researchers trace the biomarkers of interest within a longer time window.

No MeSH data available.


Related in: MedlinePlus

(a) HPLC trace of the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C (bottom), and HPLC trace of 1-Cold in water (top). (b) SEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C. (c) TEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C.
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Figure 2: (a) HPLC trace of the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C (bottom), and HPLC trace of 1-Cold in water (top). (b) SEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C. (c) TEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C.

Mentions: All the starting materials were obtained from Adamas or Sangon Biotech. Commercially available reagents were used without further purification, unless noted otherwise. All chemicals were reagent grade or better. Furin was purchased from Biolabs (2,000 U mL-1, one unit (U) corresponds to the amount of furin that releases 1 pmol of methylcoumarinamide (MCA) from the fluorogenic peptide Boc-RVRR-MCA (Bachem) in one minute at 30 °C). 1HNMR spectra were obtained on a 300 MHz Bruker AV 300. MALDI-TOF/TOF and ESI mass spectra were obtained on a time-of-flight Ultrflex II mass spectrometer (Bruker Daltonics) and on a Finnigan LCQ Advantage ion trap mass spectrometer (ThermoFisher Corporation) equipped with a standard ESI source, respectively. High performance liquid chromatography (HPLC) analyses were performed on an Agilent 1200 HPLC system equipped with a G1322A pump and in-line diode array UV detector and a YMC-Pack ODS-AM column with CH3CN (0.1% of TFA) and water (0.1% of TFA) as the eluent. Semipreparative HPLC for radiosyntheses were performed on a Waters preparative chromatography packed with waters 2998 photodiode array detector and Bioscan flow-count equipped with C18 column (5 µm, 10 mm × 250 mm, Wuxi Chrom-matrix Bio-Technology Co., Ltd. China) with CH3CN (0.1% of TFA) and water (0.1% of TFA) as the eluent. Dynamic light scattering (DLS) was measured on a Zeta Sizer Nano Series (Malvern Instruments). Transmission electron micrograph (TEM) images in Figure 2 were obtained on a JEOL 2100 electron microscope, operating at 200 kV. The cryo-dried samples were prepared as following: a copper grid coated with carbon was dipped into the suspension solvent and placed into a vial, which was plunged into liquid nitrogen until no bubbles were apparent. Then water was removed from the frozen specimen by a freeze-drier. TEM images in Figure 3 were obtained on a FEI Techni 12 electron microscope, operating at 120 kV. UV-vis absorbance spectra were recorded on a lambda 25 UV-visible spectrophotometer (PerkinElmer, America) at room temperature. Fluorescence spectra were recorded on a F-4600 fluorescence spectrophotometer (Hitachi High-Technologies Corporation, Japan) with excitation wavelength set to 320 nm. [18F]-fluoride was prepared by medical cyclotron (Sumitomo HM-7, Japan). Micro-PET imaging was performed on an Inveon scanner (Siemens, Germany). The radionuclide activity meter was curiementor-3 (PTW, Germany).


Enzyme-Controlled Intracellular Self-Assembly of (18)F Nanoparticles for Enhanced MicroPET Imaging of Tumor.

Liu Y, Miao Q, Zou P, Liu L, Wang X, An L, Zhang X, Qian X, Luo S, Liang G - Theranostics (2015)

(a) HPLC trace of the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C (bottom), and HPLC trace of 1-Cold in water (top). (b) SEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C. (c) TEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4508496&req=5

Figure 2: (a) HPLC trace of the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C (bottom), and HPLC trace of 1-Cold in water (top). (b) SEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C. (c) TEM images of the nanoparticles in the incubation mixture of 1-Cold at 100 µM after 4 h incubation with 1 nmol/U of furin at 37 °C.
Mentions: All the starting materials were obtained from Adamas or Sangon Biotech. Commercially available reagents were used without further purification, unless noted otherwise. All chemicals were reagent grade or better. Furin was purchased from Biolabs (2,000 U mL-1, one unit (U) corresponds to the amount of furin that releases 1 pmol of methylcoumarinamide (MCA) from the fluorogenic peptide Boc-RVRR-MCA (Bachem) in one minute at 30 °C). 1HNMR spectra were obtained on a 300 MHz Bruker AV 300. MALDI-TOF/TOF and ESI mass spectra were obtained on a time-of-flight Ultrflex II mass spectrometer (Bruker Daltonics) and on a Finnigan LCQ Advantage ion trap mass spectrometer (ThermoFisher Corporation) equipped with a standard ESI source, respectively. High performance liquid chromatography (HPLC) analyses were performed on an Agilent 1200 HPLC system equipped with a G1322A pump and in-line diode array UV detector and a YMC-Pack ODS-AM column with CH3CN (0.1% of TFA) and water (0.1% of TFA) as the eluent. Semipreparative HPLC for radiosyntheses were performed on a Waters preparative chromatography packed with waters 2998 photodiode array detector and Bioscan flow-count equipped with C18 column (5 µm, 10 mm × 250 mm, Wuxi Chrom-matrix Bio-Technology Co., Ltd. China) with CH3CN (0.1% of TFA) and water (0.1% of TFA) as the eluent. Dynamic light scattering (DLS) was measured on a Zeta Sizer Nano Series (Malvern Instruments). Transmission electron micrograph (TEM) images in Figure 2 were obtained on a JEOL 2100 electron microscope, operating at 200 kV. The cryo-dried samples were prepared as following: a copper grid coated with carbon was dipped into the suspension solvent and placed into a vial, which was plunged into liquid nitrogen until no bubbles were apparent. Then water was removed from the frozen specimen by a freeze-drier. TEM images in Figure 3 were obtained on a FEI Techni 12 electron microscope, operating at 120 kV. UV-vis absorbance spectra were recorded on a lambda 25 UV-visible spectrophotometer (PerkinElmer, America) at room temperature. Fluorescence spectra were recorded on a F-4600 fluorescence spectrophotometer (Hitachi High-Technologies Corporation, Japan) with excitation wavelength set to 320 nm. [18F]-fluoride was prepared by medical cyclotron (Sumitomo HM-7, Japan). Micro-PET imaging was performed on an Inveon scanner (Siemens, Germany). The radionuclide activity meter was curiementor-3 (PTW, Germany).

Bottom Line: TEM images of 1-Cold-treated MDA-MB-468 cells directly uncovered that the intracellular 1-Cold-NPs were at/near the location of furin (i.e., Golgi bodies).MTT results indicated that 50 µM 1-Cold did not impose cytotoxicity to MDA-MB-468 cells up to 12 hours.Our ''smart'' probe (i.e., 1), together with the strategy of co-injection, might help researchers trace the biomarkers of interest within a longer time window.

View Article: PubMed Central - PubMed

Affiliation: 1. School of Chemical and Material Engineering, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China ; 3. Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.

ABSTRACT
Herein, we report the development of a new "smart" radioactive probe (i.e., 1) which can undergo furin-controlled condensation and self-assembly of radioactive nanoparticles (i.e., 1-NPs) in tumor cells and its application for enhanced microPET imaging of tumors in nude mice co-injected with its cold analog (i.e., 1-Cold). Furin-controlled condensation of 1-Cold and self-assembly of its nanoparticles (i.e., 1-Cold-NPs) in vitro were validated and characterized with HPLC, mass spectra, SEM, and TEM analyses. Cell uptake studies showed that both 1 and 1-Cold have good cell permeability. TEM images of 1-Cold-treated MDA-MB-468 cells directly uncovered that the intracellular 1-Cold-NPs were at/near the location of furin (i.e., Golgi bodies). MTT results indicated that 50 µM 1-Cold did not impose cytotoxicity to MDA-MB-468 cells up to 12 hours. MicroPET imaging of MDA-MB-468 tumor-bearing mice indicated that mice co-injected with 1 and 1-Cold showed higher uptake and longer attenuation of the radioactivity in tumors than those mice only injected with same dosage of 1. Tumor uptake ratios of 1 between these two groups of mice reached the maximum of 8.2 folds at 240 min post injection. Biodistribution study indicated that the uptake ratios of 1 in kidneys between these two groups continuously increased and reached 81.9 folds at 240 min post injection, suggesting the formation of radioactive NPs (i.e., 1-NPs) in MDA-MB-468 tumors of mice co-injected with 1 and 1-Cold. And the nanoparticles were slowly digested and secreted from the tumors, accumulating in the kidneys. Our ''smart'' probe (i.e., 1), together with the strategy of co-injection, might help researchers trace the biomarkers of interest within a longer time window.

No MeSH data available.


Related in: MedlinePlus