Dendrimer-based targeted delivery of an apoptotic sensor in cancer cells.
Our previous studies have demonstrated the applicability of poly(amidoamine) (PAMAM) dendrimers as a platform for the targeted delivery of chemotherapeutic drugs both in vitro and in vivo. To monitor the rate and extent of cell-killing caused by the delivered chemotherapeutic drug, we wished to analyze the degree of apoptosis in targeted cells on a real-time basis. As the apoptosis-regulating caspases are activated during the apoptotic process, several caspase-hydrolyzable, fluorescence resonance energy transfer (FRET)-based substrates have been marketed for the detection of apoptosis. However, the applicability of these agents is limited because of their nonspecificity and the consequent high background fluorescence in tissues. Here we show the synthesis, characterization, and in vitro targeting of an engineered PAMAM nanodevice in which folic acid (FA) is conjugated as the targeting molecule and a caspase-specific FRET-based agent (PhiPhiLux G1D2) is conjugated as the apoptosis-detecting agent. This conjugate specifically targets FA-receptor-positive, KB cells. In these cells, the apoptosis-inducing agent staurosporine caused a 5-fold increase in the cellular fluorescence. These results show, for the first time, the potential applicability of a targeted apoptosis-measuring nanodevice, which could be used for simultaneously monitoring the apoptotic potential of a delivered drug.
Myc
Andrzej
A
Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan, Ann Arbor, Michigan 48109, USA.
Majoros
István J
IJ
Thomas
Thommey P
TP
Baker
James R
JR
D017209:Apoptosis; D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D050091:Dendrimers / Q000737:chemistry*; D016503:Drug Delivery Systems; D004563:Electrochemistry; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D006868:Hydrolysis; D019169:Jurkat Cells; D008956:Models, Chemical; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*
D050091:Dendrimers