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Manganese-containing Prussian blue nanoparticles for imaging of pediatric brain tumors.

Dumont MF, Yadavilli S, Sze RW, Nazarian J, Fernandes R - Int J Nanomedicine (2014)

Bottom Line: Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs.We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles.Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro.

View Article: PubMed Central - PubMed

Affiliation: Sheikh Zayed Institute for Pediatric Surgical Innovation, Washington, DC, USA.

ABSTRACT
Pediatric brain tumors (PBTs) are a leading cause of death in children. For an improved prognosis in patients with PBTs, there is a critical need to develop molecularly-specific imaging agents to monitor disease progression and response to treatment. In this paper, we describe manganese-containing Prussian blue nanoparticles as agents for molecular magnetic resonance imaging (MRI) and fluorescence-based imaging of PBTs. Our core-shell nanoparticles consist of a core lattice structure that incorporates and retains paramagnetic Mn(2+) ions, and generates MRI contrast (both negative and positive). The biofunctionalized shell is comprised of fluorescent avidin, which serves the dual purpose of enabling fluorescence imaging and functioning as a platform for the attachment of biotinylated ligands that target PBTs. The surfaces of our nanoparticles are modified with biotinylated antibodies targeting neuron-glial antigen 2 or biotinylated transferrin. Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs. We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles. Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro. Finally, we demonstrate the potential of these nanoparticles as PBT imaging agents by measuring their organ and brain biodistribution in an orthotopic mouse model of PBTs using ex vivo fluorescence imaging.

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Flow cytometric analysis of the specificity of biofunctionalized MnPB nanoparticles for PBT cells.Notes: (A) Representative histograms of cell count plotted against Alexa Fluor 488 detection levels for BSG D10 cells treated with MnPB-A488 (no antibody, red line), MnPB-A488-AbC (blue line), and MnPB-A488-ANG2 (black line), and stained with 7-AAD. (B) Percentage Alexa Fluor 488-positive cells (fluorescence intensity cutoff 50) cells for BSG D10 treated with MnPB-A488-ANG2, MnPB-A488-AbC, or MnPB-A488. **P<0.05.Abbreviations: BSG, brainstem glioma; 7-AAD, 7-aminoactinomycin D; MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2; PBT, pediatric brain tumor; au, arbitrary units.
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f7-ijn-9-2581: Flow cytometric analysis of the specificity of biofunctionalized MnPB nanoparticles for PBT cells.Notes: (A) Representative histograms of cell count plotted against Alexa Fluor 488 detection levels for BSG D10 cells treated with MnPB-A488 (no antibody, red line), MnPB-A488-AbC (blue line), and MnPB-A488-ANG2 (black line), and stained with 7-AAD. (B) Percentage Alexa Fluor 488-positive cells (fluorescence intensity cutoff 50) cells for BSG D10 treated with MnPB-A488-ANG2, MnPB-A488-AbC, or MnPB-A488. **P<0.05.Abbreviations: BSG, brainstem glioma; 7-AAD, 7-aminoactinomycin D; MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2; PBT, pediatric brain tumor; au, arbitrary units.

Mentions: We quantitatively measured the binding of MnPB-A488-ANG2 to BSG D10 using flow cytometry. We separately added MnPB-A488-ANG2 and controls (MnPB-A488-AbC and MnPB-A488) to cultures of BSG D10 and measured the distribution of the cell populations that were fluorescently labeled (denoted as Alexa Fluor 488-positive; Figures 7A and S7). The MnPB-A488-ANG2 nanoparticles specifically bound to the BSG D10 cells (81%±4% Alexa Fluor 488-positive; Figure 7B) compared with MnPB-A488-AbC (23%±2% Alexa Fluor 488-positive; Figure 7B) and MnPB-A488 (22%±3% Alexa Fluor 488-positive; Figure 7B) where the nanoparticles showed background levels of binding to BSG D10 cells. Taken together, these results demonstrated the ability of our nanoparticles (with antibody ANG2) to molecularly target BSG D10 cells effectively using fluorescence imaging.


Manganese-containing Prussian blue nanoparticles for imaging of pediatric brain tumors.

Dumont MF, Yadavilli S, Sze RW, Nazarian J, Fernandes R - Int J Nanomedicine (2014)

Flow cytometric analysis of the specificity of biofunctionalized MnPB nanoparticles for PBT cells.Notes: (A) Representative histograms of cell count plotted against Alexa Fluor 488 detection levels for BSG D10 cells treated with MnPB-A488 (no antibody, red line), MnPB-A488-AbC (blue line), and MnPB-A488-ANG2 (black line), and stained with 7-AAD. (B) Percentage Alexa Fluor 488-positive cells (fluorescence intensity cutoff 50) cells for BSG D10 treated with MnPB-A488-ANG2, MnPB-A488-AbC, or MnPB-A488. **P<0.05.Abbreviations: BSG, brainstem glioma; 7-AAD, 7-aminoactinomycin D; MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2; PBT, pediatric brain tumor; au, arbitrary units.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043710&req=5

f7-ijn-9-2581: Flow cytometric analysis of the specificity of biofunctionalized MnPB nanoparticles for PBT cells.Notes: (A) Representative histograms of cell count plotted against Alexa Fluor 488 detection levels for BSG D10 cells treated with MnPB-A488 (no antibody, red line), MnPB-A488-AbC (blue line), and MnPB-A488-ANG2 (black line), and stained with 7-AAD. (B) Percentage Alexa Fluor 488-positive cells (fluorescence intensity cutoff 50) cells for BSG D10 treated with MnPB-A488-ANG2, MnPB-A488-AbC, or MnPB-A488. **P<0.05.Abbreviations: BSG, brainstem glioma; 7-AAD, 7-aminoactinomycin D; MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2; PBT, pediatric brain tumor; au, arbitrary units.
Mentions: We quantitatively measured the binding of MnPB-A488-ANG2 to BSG D10 using flow cytometry. We separately added MnPB-A488-ANG2 and controls (MnPB-A488-AbC and MnPB-A488) to cultures of BSG D10 and measured the distribution of the cell populations that were fluorescently labeled (denoted as Alexa Fluor 488-positive; Figures 7A and S7). The MnPB-A488-ANG2 nanoparticles specifically bound to the BSG D10 cells (81%±4% Alexa Fluor 488-positive; Figure 7B) compared with MnPB-A488-AbC (23%±2% Alexa Fluor 488-positive; Figure 7B) and MnPB-A488 (22%±3% Alexa Fluor 488-positive; Figure 7B) where the nanoparticles showed background levels of binding to BSG D10 cells. Taken together, these results demonstrated the ability of our nanoparticles (with antibody ANG2) to molecularly target BSG D10 cells effectively using fluorescence imaging.

Bottom Line: Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs.We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles.Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro.

View Article: PubMed Central - PubMed

Affiliation: Sheikh Zayed Institute for Pediatric Surgical Innovation, Washington, DC, USA.

ABSTRACT
Pediatric brain tumors (PBTs) are a leading cause of death in children. For an improved prognosis in patients with PBTs, there is a critical need to develop molecularly-specific imaging agents to monitor disease progression and response to treatment. In this paper, we describe manganese-containing Prussian blue nanoparticles as agents for molecular magnetic resonance imaging (MRI) and fluorescence-based imaging of PBTs. Our core-shell nanoparticles consist of a core lattice structure that incorporates and retains paramagnetic Mn(2+) ions, and generates MRI contrast (both negative and positive). The biofunctionalized shell is comprised of fluorescent avidin, which serves the dual purpose of enabling fluorescence imaging and functioning as a platform for the attachment of biotinylated ligands that target PBTs. The surfaces of our nanoparticles are modified with biotinylated antibodies targeting neuron-glial antigen 2 or biotinylated transferrin. Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs. We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles. Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro. Finally, we demonstrate the potential of these nanoparticles as PBT imaging agents by measuring their organ and brain biodistribution in an orthotopic mouse model of PBTs using ex vivo fluorescence imaging.

Show MeSH
Related in: MedlinePlus