Chitosan-based therapeutic nanoparticles for combination gene therapy and gene silencing of in vitro cell lines relevant to type 2 diabetes

详细信息    查看全文

• 作者：Myriam  ; Jean¹ ; Mohamad  ; Alameh¹ ; Diogo  ; De Jesus ; Marc  ; Thibault ; Marc  ; Lavertu ; Vincent  ; Darras ; Monica  ; Nelea ; Michael D.  ; Buschmann ; Abderrazzak  ; Merzouki  ; ;   ; ^{abderrazzak.merzouki@polymtl.ca}
• 关键词：Type 2 diabetes ; Glucagon like peptide 1 ; Dipeptydil-peptidase IV ; siRNA ; Chitosan ; Gene delivery system
• 刊名：European Journal of Pharmaceutical Sciences
• 出版年：2012
• 出版时间：23 January 2012
• 年：2012
• 卷：45
• 期：1-2
• 页码：138-149
• 全文大小：1217 K

文摘

Glucagon like peptide 1 (GLP-1), a blood glucose homeostasis modulating incretin, has been proposed for the treatment of type 2 diabetes mellitus (T2DM). However, native GLP-1 pharmacokinetics reveals low bioavailability due to degradation by the ubiquitous dipeptydil peptidase IV (DPP-IV) endoprotease. In this study, the glucosamine-based polymer chitosan was used as a cationic polymer-based in vitro delivery system for GLP-1, DPP-IV resistant GLP-1 analogues and siRNA targeting DPP-IV mRNA. We found chitosans to form spherical nanocomplexes with these nucleic acids, generating two distinct non-overlapping size ranges of 141?83 nm and 68?29 nm for plasmid and siRNA, respectively. The low molecular weight high DDA chitosan 92-10-5 (degree of deacetylation, molecular weight and N:P ratio (DDA¨CMn¨CN:P)) showed the highest plasmid DNA transfection efficiency in HepG2 and Caco-2 cell lines when compared to 80?0?0 and 80?0? chitosans. Recombinant native GLP-1 protein levels in media of transfected cells reached 23 ng/L while our DPP-IV resistant analogues resulted in a fivefold increase of GLP-1 protein levels (115 ng/L) relative to native GLP-1, and equivalent to the Lipofectamine positive control. We also found that all chitosan¨CDPP-IV siRNA nanocomplexes were capable of DPP-IV silencing, with 92?0? being significantly more effective in abrogating enzymatic activity of DPP-IV in media of silenced cells, and with no apparent cytotoxicity. These results indicate that specific chitosan formulations may be effectively used for the delivery of plasmid DNA and siRNA in a combination therapy of type 2 diabetes.