An inhalable β₂-adrenoceptor ligand-directed guanidinylated chitosan carrier for targeted delivery of siRNA to lung.
SiRNA-based strategies appear to be an exciting new approach for the treatment of respiratory diseases. To extrapolate siRNA-mediated interventions from bench to bedside in this area, several aspects have to be jointly considered, including a safe and efficient gene carrier with pulmonary deposition efficiency, as well as in vivo method for siRNA/nanoparticles delivery. Accordingly, in this work, (i) a non-viral DNA vector, guanidinylated chitosan (GCS) that has been developed in our previous study [X.Y. Zhai, P. Sun, Y.F. Luo, C.N. Ma, J. Xu, W.G. Liu, 2011], was tested for siRNA delivery. We demonstrated that GCS was able to completely condense siRNA at weight ratio 40:1, forming nanosize particles of diameter ~100 nm, 15 mV in surface potential. Guanidinylation of chitosan not only decreased the cytotoxicity but also facilitated cellular internalization of siRNA nanoparticles, leading to an enhanced gene-silencing efficiency compared to the pristine chitosan (CS). (ii) We chemically coupled salbutamol, a β(2)-adrenoceptor agonist, to GCS (SGCS), which successfully improved targeting specificity of the green fluorescent protein (GFP)-siRNA carrier to lung cells harbored with β(2)-adrenergic receptor, and remarkably enhanced the efficacy of gene silence in vitro and in the lung of enhanced green fluorescent protein (EGFP)-transgenic mice in vivo. (iii) It was proved that this chitosan-based polymer was able to provide both the pDNA and siRNA with the protection against destructive shear forces generated by the mesh-based nebulizers. Aerosol treatment improved the nanoparticle size distribution, which should be in favor of enhancing the transfection efficiency. We suggest a potential application of the chitosan-derived nanodelivery vehicle (SGCS) in RNA interference therapy for lung diseases via aerosol inhalation.
State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou 510120, Guangdong Province, China.
Yongfeng
Y
Luo
Xinyun
X
Zhai
Chaonan
C
Ma
Peng
P
Sun
Zhiping
Z
Fu
Wenguang
W
Liu
Jun
J
Xu
D058666:Adrenergic beta-2 Receptor Agonists / Q000737:chemistry / Q000378:metabolism; D000420:Albuterol / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D048271:Chitosan / Q000031:analogs & derivatives* / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry* / Q000378:metabolism; D016503:Drug Delivery Systems; D049452:Green Fluorescent Proteins / Q000235:genetics; D019791:Guanidine / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009330:Nebulizers and Vaporizers; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism*
D058666:Adrenergic beta-2 Receptor Agonists; D004337:Drug Carriers; D034741:RNA, Small Interfering; D018343:Receptors, Adrenergic, beta-2; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D048271:Chitosan; D019791:Guanidine; D000420:Albuterol