Chitosan and depolymerized chitosan oligomers as condensing carriers for in vivo plasmid delivery.
Chitosan is a polysaccharide that demonstrates much potential as a gene delivery system. The ability of a commercially available chitosan and depolymerized chitosan oligomers to condense plasmid was determined using TEM and microtitration calorimetry, while the diameter and stability of the resultant complexes were measured using laser light scattering. Selected complexes were physically stable to challenge with both serum and salt solutions. Parameters such as chitosan molecular weight, plasmid concentration and charge ratio influenced such stability. The effect of including a pH-sensitive endosomolytic peptide on the physicochemical properties of the complex was determined. The presence of a pH-sensitive endosomolytic peptide enhanced the levels of reporter gene expression in Cos-1 cells 4-fold. A selected complex containing a lytic peptide was administered in the upper small intestine and colon of rabbits, and reporter gene expression was measured in defined intestinal tissues. Reporter gene expression was enhanced in defined intestinal tissues, although levels of expression remained low. The combination of strong complex stability and low in vivo expression levels suggest that uptake and/or decomplexation, but not endosomal release, may be the critical rate-limiting steps in the uptake process.
MacLaughlin
F C
FC
GeneMedicine Inc., 8301 New Trails Drive, The Woodlands, TX 77381-4248, USA.
Mumper
R J
RJ
Wang
J
J
Tagliaferri
J M
JM
Gill
I
I
Hinchcliffe
M
M
Rolland
A P
AP
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002686:Chitin / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry; D048271:Chitosan; D003106:Colon / Q000502:physiology; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004355:Drug Stability; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006863:Hydrogen-Ion Concentration; D007421:Intestine, Small / Q000502:physiology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000008:administration & dosage / Q000494:pharmacology; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011817:Rabbits; D014162:Transfection
D010455:Peptides; D002686:Chitin; D004247:DNA; D048271:Chitosan