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MUC1-Targeted Cancer Cell Photothermal Ablation Using Bioinspired Gold Nanorods.

Zelasko-Leon DC, Fuentes CM, Messersmith PB - PLoS ONE (2015)

Bottom Line: MUC1-BSA-PD-NRs substantially decreased cell viability in photoirradiated MCF-7 cell lines vs.Agents exhibited no cytotoxicity in the absence of photothermal treatment.The facile nature of the coating method, combined with targeting and photoablation efficacy, are attractive features of these candidate cancer nanotherapeutics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Engineering, Chemistry of Life Processes Institute, Northwestern University, Evanston, Illinois, United States of America; Department of Bioengineering, University of California, Berkeley, Berkeley, California, United States of America.

ABSTRACT
Recent studies have highlighted the overexpression of mucin 1 (MUC1) in various epithelial carcinomas and its role in tumorigenesis. These mucins present a novel targeting opportunity for nanoparticle-mediated photothermal cancer treatments due to their unique antenna-like extracellular extension. In this study, MUC1 antibodies and albumin were immobilized onto the surface of gold nanorods using a "primer" of polydopamine (PD), a molecular mimic of catechol- and amine-rich mussel adhesive proteins. PD forms an adhesive platform for the deposition of albumin and MUC1 antibodies, achieving a surface that is stable, bioinert and biofunctional. Two-photon luminescence confocal and darkfield scattering imaging revealed targeting of MUC1-BSA-PD-NRs to MUC1+ MCF-7 breast cancer and SCC-15 squamous cell carcinoma cells lines. Treated cells were exposed to a laser encompassing the near-infrared AuNR longitudinal surface plasmon and assessed for photothermal ablation. MUC1-BSA-PD-NRs substantially decreased cell viability in photoirradiated MCF-7 cell lines vs. MUC1- MDA-MB-231 breast cancer cells (p < 0.005). Agents exhibited no cytotoxicity in the absence of photothermal treatment. The facile nature of the coating method, combined with targeting and photoablation efficacy, are attractive features of these candidate cancer nanotherapeutics.

No MeSH data available.


Related in: MedlinePlus

Anti-MUC1 conjugation efficiency.(A) LSPR peak red shifting is observed with sequential BSA and anti-MUC1 modifications. (B) ELISA reveals total incorporation of MUC1-N and—C antibodies was achieved (* p < 0.05) only in PD-primed NRs additionally stabilized via a BSA layer. Near 100% loading in MUC1-C-BSA-NR samples has no practical significance as antibody addition induced complete NR aggregation (#). To quantify the differential MUC1 expression profiles in our cell lines, cells were treated with PE-conjugated MUC1 antibodies and analyzed via flow cytometry for their mean fluorescence intensities (MFI) against cells labeled with isotype controls. Both MCF-7 (MUC1++) and SCC15 (MUC1+) cells demonstrated an enhanced MFI in agreement with literature reports [16, 67, 68]. As expected, MUC1-deficient MDA-MB-231 (MUC1-) cells did not show an appreciable increase in anti-MUC1 PE labeling (Fig F in S1 File).
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pone.0128756.g003: Anti-MUC1 conjugation efficiency.(A) LSPR peak red shifting is observed with sequential BSA and anti-MUC1 modifications. (B) ELISA reveals total incorporation of MUC1-N and—C antibodies was achieved (* p < 0.05) only in PD-primed NRs additionally stabilized via a BSA layer. Near 100% loading in MUC1-C-BSA-NR samples has no practical significance as antibody addition induced complete NR aggregation (#). To quantify the differential MUC1 expression profiles in our cell lines, cells were treated with PE-conjugated MUC1 antibodies and analyzed via flow cytometry for their mean fluorescence intensities (MFI) against cells labeled with isotype controls. Both MCF-7 (MUC1++) and SCC15 (MUC1+) cells demonstrated an enhanced MFI in agreement with literature reports [16, 67, 68]. As expected, MUC1-deficient MDA-MB-231 (MUC1-) cells did not show an appreciable increase in anti-MUC1 PE labeling (Fig F in S1 File).

Mentions: Antibody conjugation was qualitatively confirmed via SDS-PAGE (Fig E in S1 File) and optical spectroscopy of LSPR red shift (Fig 3A). To quantify the degree of MUC1 antibody conjugation to modified AuNRs, a MUC1 ELISA was designed. Goat anti-mouse coated microplates were used to capture unbound mouse anti-human MUC1 antibodies sourced from AuNR sample supernatants to calculate the amount of depleted antibody from known starting concentrations. The results of the ELISA indicate that antibody was completely incorporated in BSA-PD-NR samples whereas BSA-NR modified with MUC1-N (BSA-NR N) incorporated only 72% of the available antibody under the same conditions (p < 0.05)–affording a facile and versatile coupling strategy for cancer targeting applications. Treatment of BSA-NR with MUC1-C antibodies (BSA-NR C) resulted in the immediate aggregation of the sample (Fig 3B), further supporting the potential covalent nature of the BSA-PD coating and its robust ability to survive in physiologically relevant environments.


MUC1-Targeted Cancer Cell Photothermal Ablation Using Bioinspired Gold Nanorods.

Zelasko-Leon DC, Fuentes CM, Messersmith PB - PLoS ONE (2015)

Anti-MUC1 conjugation efficiency.(A) LSPR peak red shifting is observed with sequential BSA and anti-MUC1 modifications. (B) ELISA reveals total incorporation of MUC1-N and—C antibodies was achieved (* p < 0.05) only in PD-primed NRs additionally stabilized via a BSA layer. Near 100% loading in MUC1-C-BSA-NR samples has no practical significance as antibody addition induced complete NR aggregation (#). To quantify the differential MUC1 expression profiles in our cell lines, cells were treated with PE-conjugated MUC1 antibodies and analyzed via flow cytometry for their mean fluorescence intensities (MFI) against cells labeled with isotype controls. Both MCF-7 (MUC1++) and SCC15 (MUC1+) cells demonstrated an enhanced MFI in agreement with literature reports [16, 67, 68]. As expected, MUC1-deficient MDA-MB-231 (MUC1-) cells did not show an appreciable increase in anti-MUC1 PE labeling (Fig F in S1 File).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493038&req=5

pone.0128756.g003: Anti-MUC1 conjugation efficiency.(A) LSPR peak red shifting is observed with sequential BSA and anti-MUC1 modifications. (B) ELISA reveals total incorporation of MUC1-N and—C antibodies was achieved (* p < 0.05) only in PD-primed NRs additionally stabilized via a BSA layer. Near 100% loading in MUC1-C-BSA-NR samples has no practical significance as antibody addition induced complete NR aggregation (#). To quantify the differential MUC1 expression profiles in our cell lines, cells were treated with PE-conjugated MUC1 antibodies and analyzed via flow cytometry for their mean fluorescence intensities (MFI) against cells labeled with isotype controls. Both MCF-7 (MUC1++) and SCC15 (MUC1+) cells demonstrated an enhanced MFI in agreement with literature reports [16, 67, 68]. As expected, MUC1-deficient MDA-MB-231 (MUC1-) cells did not show an appreciable increase in anti-MUC1 PE labeling (Fig F in S1 File).
Mentions: Antibody conjugation was qualitatively confirmed via SDS-PAGE (Fig E in S1 File) and optical spectroscopy of LSPR red shift (Fig 3A). To quantify the degree of MUC1 antibody conjugation to modified AuNRs, a MUC1 ELISA was designed. Goat anti-mouse coated microplates were used to capture unbound mouse anti-human MUC1 antibodies sourced from AuNR sample supernatants to calculate the amount of depleted antibody from known starting concentrations. The results of the ELISA indicate that antibody was completely incorporated in BSA-PD-NR samples whereas BSA-NR modified with MUC1-N (BSA-NR N) incorporated only 72% of the available antibody under the same conditions (p < 0.05)–affording a facile and versatile coupling strategy for cancer targeting applications. Treatment of BSA-NR with MUC1-C antibodies (BSA-NR C) resulted in the immediate aggregation of the sample (Fig 3B), further supporting the potential covalent nature of the BSA-PD coating and its robust ability to survive in physiologically relevant environments.

Bottom Line: MUC1-BSA-PD-NRs substantially decreased cell viability in photoirradiated MCF-7 cell lines vs.Agents exhibited no cytotoxicity in the absence of photothermal treatment.The facile nature of the coating method, combined with targeting and photoablation efficacy, are attractive features of these candidate cancer nanotherapeutics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Engineering, Chemistry of Life Processes Institute, Northwestern University, Evanston, Illinois, United States of America; Department of Bioengineering, University of California, Berkeley, Berkeley, California, United States of America.

ABSTRACT
Recent studies have highlighted the overexpression of mucin 1 (MUC1) in various epithelial carcinomas and its role in tumorigenesis. These mucins present a novel targeting opportunity for nanoparticle-mediated photothermal cancer treatments due to their unique antenna-like extracellular extension. In this study, MUC1 antibodies and albumin were immobilized onto the surface of gold nanorods using a "primer" of polydopamine (PD), a molecular mimic of catechol- and amine-rich mussel adhesive proteins. PD forms an adhesive platform for the deposition of albumin and MUC1 antibodies, achieving a surface that is stable, bioinert and biofunctional. Two-photon luminescence confocal and darkfield scattering imaging revealed targeting of MUC1-BSA-PD-NRs to MUC1+ MCF-7 breast cancer and SCC-15 squamous cell carcinoma cells lines. Treated cells were exposed to a laser encompassing the near-infrared AuNR longitudinal surface plasmon and assessed for photothermal ablation. MUC1-BSA-PD-NRs substantially decreased cell viability in photoirradiated MCF-7 cell lines vs. MUC1- MDA-MB-231 breast cancer cells (p < 0.005). Agents exhibited no cytotoxicity in the absence of photothermal treatment. The facile nature of the coating method, combined with targeting and photoablation efficacy, are attractive features of these candidate cancer nanotherapeutics.

No MeSH data available.


Related in: MedlinePlus