Combining Stimulus-Triggered Release and Active Targeting Strategies Improves Cytotoxicity of Cytochrome c Nanoparticles in Tumor Cells.
Proteins often possess highly specific biological activities that make them potential therapeutics, but their physical and chemical instabilities during formulation, storage, and delivery have limited their medical use. Therefore, engineering of nanosized vehicles to stabilize protein therapeutics and to allow for targeted treatment of complex diseases, such as cancer, is of considerable interest. A micelle-like nanoparticle (NP) was designed for both, tumor targeting and stimulus-triggered release of the apoptotic protein cytochrome c (Cyt c). This system is composed of a Cyt c NP stabilized by a folate-receptor targeting amphiphilic copolymer (FA-PEG-PLGA) attached to Cyt c through a redox-sensitive bond. FA-PEG-PLGA-S-S-Cyt c NPs exhibited excellent stability under extracellular physiological conditions, whereas once in the intracellular reducing environment, Cyt c was released from the conjugate. Under the same conditions, the folate-decorated NP reduced folate receptor positive HeLa cell viability to 20%, while the same complex without FA only reduced it to 80%. Confocal microscopy showed that the FA-PEG-PLGA-S-S-Cyt c NPs were internalized by HeLa cells and were capable of endosomal escape. The specificity of the folate receptor-mediated internalization was confirmed by the lack of uptake by two folate receptor deficient cell lines: A549 and NIH-3T3. Finally, the potential as antitumor therapy of our folate-decorated Cyt c-based NPs was confirmed with an in vivo brain tumor model. In conclusion, we were able to create a stable, selective, and smart nanosized Cyt c delivery system.
Department of Biology, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Moraima
M
Morales-Cruz
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Alejandra
A
Cruz-Montañez
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Cindy M
CM
Figueroa
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Tania
T
González-Robles
Department of Biochemistry, Universidad Central del Caribe, School of Medicine , Bayamón 00560, Puerto Rico.
Josue
J
Davila
Department of Physiology, Universidad Central del Caribe, School of Medicine , Bayamón 00560, Puerto Rico.
Mikhail
M
Inyushin
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Sergio A
SA
Loza-Rosas
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Anna M
AM
Molina
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Laura
L
Muñoz-Perez
Department of Biochemistry, Universidad Central del Caribe, School of Medicine , Bayamón 00560, Puerto Rico.
Lilia Y
LY
Kucheryavykh
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Arthur D
AD
Tinoco
Department of Chemistry, University of Puerto Rico , Río Piedras Campus, San Juan 00931, Puerto Rico.
Kai
K
Griebenow
D000072283:A549 Cells; D000818:Animals; D017209:Apoptosis; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism*; D004337:Drug Carriers / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods; D005910:Glioma / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008823:Micelles; D041681:NIH 3T3 Cells; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry
D004337:Drug Carriers; D008823:Micelles; D011108:Polymers; D045304:Cytochromes c
active targeting; biodegradable polymer; drug delivery; nanoparticle; protein drug; triggered release