Poly(glycoamidoamine)s for gene delivery. structural effects on cellular internalization, buffering capacity, and gene expression.
The study of polymeric nucleic acid delivery vehicles has recently grown because of their promise for many biomedical applications. In an effort to understand how the chemical traits of polymers affect the biological mechanisms of nucleic acid delivery, we have calculated the buffering capacity in the physiological pH range of a series of 10 poly(glycoamidoamine)s with systematic structural variations in the amine stoichiometry (from 1 to 4), carbohydrate moiety (d-glucarate or l-tartarate), and amine spacer (ethylene or butylene) within their repeat units. In addition, we have compared the buffering capacity of these polymeric vectors to their polyplex (polymer-DNA complex) stability, cellular internalization, and gene expression profiles to understand the parameters that are important for increasing gene delivery efficiency. The results indicate that the buffering capacity is not always the primary characteristic that determines the gene delivery efficiency for all the poly(glycoamidoamine)s. We have found that the buffering capacity may affect the gene delivery efficiency only when analogous structures containing the same number of amines but different carbohydrates are compared. We reveal that the cellular internalization is the key step in the gene delivery process with systems containing different amine stoichiometry. Also, increasing the number of methylene groups between the secondary amines increases toxicity to a large degree. This systematic and heuristic approach of studying the correlations between structural variables and gene delivery efficiency will facilitate the development of effective synthetic vectors for specific nucleic acid delivery applications.
Liu
Yemin
Y
Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172, USA.
Reineke
Theresa M
TM
D000577:Amides / Q000737:chemistry*; D000588:Amines / Q000737:chemistry*; D000818:Animals; D002021:Buffers; D002241:Carbohydrates / Q000737:chemistry; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D015870:Gene Expression / Q000187:drug effects; D006031:Glycosylation; D006493:Heparin / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D008970:Molecular Weight; D017711:Nonlinear Dynamics; D010957:Plasmids / Q000235:genetics; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry* / Q000633:toxicity; D017186:Titrimetry; D014162:Transfection / Q000379:methods*
D000577:Amides; D000588:Amines; D002021:Buffers; D002241:Carbohydrates; D011108:Polymers; D006493:Heparin