TAT-LHRH conjugated low molecular weight chitosan as a gene carrier specific for hepatocellular carcinoma cells

Abstract

To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC). TLC/DNA nanoparticles (TLCDNPs) were characterized by agarose gel retardation, atomic force microscopy, and dynamic light scattering analysis. In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System. The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70–85 nm and zeta potential of +30 mV and were relatively stable in solution. The in vitro study demonstrated TLC was highly selective for hepatoma cells and essentially nontoxic.

Biodistribution of TAT‐LHRH conjugated chitosan/DNA nanoparticles in the mice bearing hepatoma xenografts

“…Because chitosan/DNA nanoparticles are unstable under physiological condition, they would rapidly disassemble and release the DNA once getting into circulation, which led to a marginal delayed renal clearance as compared with the naked DNA. However, as revealed in our previous report, TLC was able to form nanoscaled polyplexes with DNA that remained relatively stable in solution, and thus could prevent the rapid clearance of DNA through kidney. This might also hint that TLCDNPs had a longer circulation time as compared with chitosan/DNA nanoparticles, and this postulation would draw further support from the percentage data and the phenomenon that a rebound in the DNA accumulation in all the organs tested was observed at day 7.…”

“…To improve its tumor targeting capability and gene delivery efficiency, we developed a novel gene carrier by conjugating low molecular weight chitosan with transactivator of transcription (TAT) peptide and luteinizing hormone‐releasing hormone (LHRH) analog, combining the cell penetrating capability of TAT, cancer targeting specificity of LHRH and the DNA condensing power of chitosan. The resultant TAT‐LHRH‐chitosan conjugate (TLC) was demonstrated to have stronger DNA condensing power than unmodified chitosan, and form stable nanoscaled complexes with DNA, which was highly selective for hepatoma cells and essentially nontoxic as evaluated in vitro . However, it remains unclear whether TLC can deliver the genes to the target organs and tissues in vivo , which is one of the critical features determining their medical application potential.…”

“…The resultant TAT-LHRH-chitosan conjugate (TLC) was demonstrated to have stronger DNA condensing power than unmodified chitosan, and form stable nanoscaled complexes with DNA, which was highly selective for hepatoma cells and essentially nontoxic as evaluated in vitro. 34 However, it remains unclear whether TLC can deliver the genes to the target organs and tissues in vivo, which is one of the critical features determining their medical application potential.…”

Cell Penetrating Peptide Conjugated Chitosan for Enhanced Delivery of Nucleic Acid

“…To improve the transfection effectiveness and specificity, a bifunctional peptide of TAT along with luteinizing hormone-releasing hormone (LHRH) was chemically conjugated to low molecular weight chitosan, to create a TAT-LHRH-chitosan conjugate (TLC) [ 131 ]. The TLC showed stronger pDNA condensing capacity compared to unmodified chitosan and formed stable nanoscale TLC/pDNA polyplexes (70–85 nm) with a net positive surface charge of approximately +30 mV.…”

CD147 monoclonal antibody mediated by chitosan nanoparticles loaded with α-hederin enhances antineoplastic activity and cellular uptake in liver cancer cells

“…Silymarin, a hepatoprotective drug 16 , could be entrapped into CS through ionic gelation for passive targeting delivery. As a biodegradable material, CS can encapsulate antigen or DNA to protect them from damage or form complexes with DNA for gene delivery 17 18 19 20 . As a siRNA delivery nanocarrier, the transfection efficiency could be as high as 89% 21 .…”