Neurotensin-Conjugated Reduced Graphene Oxide with Multi-Stage Near-Infrared-Triggered Synergic Targeted Neuron Gene Transfection In Vitro and In Vivo for Neurodegenerative Disease Therapy.
Delivery efficiency with gene transfection is a pivotal point in achieving maximized therapeutic efficacy and has been an important challenge with central nervous system (CNS) diseases. In this study, neurotensin (NT, a neuro-specific peptide)-conjugated polyethylenimine (PEI)-modified reduced graphene oxide (rGO) nanoparticles with precisely controlled two-stage near-infrared (NIR)-laser photothermal treatment to enhance the ability to target neurons and achieve high gene transfection in neurons. First-stage NIR laser irradiation on the cells with nanoparticles attached on the surface can increase the permeability of the cell membrane, resulting in an apparent increase in cellular uptake compared to untreated cells. In addition, second-stage NIR laser irradiation on the cells with nanoparticles inside can further induce endo/lysosomal cavitation, which not only helps nanoparticles escape from endo/lysosomes but also prevents plasmid DNA (pDNA) from being digested by DNase I. At least double pDNA amount can be released from rGO-PEI-NT/pDNA under NIR laser trigger release compared to natural release. Moreover, in vitro differentiated PC-12 cell and in vivo mice (C57BL/6) brain transfection experiments have demonstrated the highest transfection efficiency occurring when NT modification is combined with external multi-stage stimuli-responsive NIR laser treatment. The combination of neuro-specific targeting peptide and external NIR-laser-triggered aid provides a nanoplatform for gene therapy in CNS diseases.
Department of Materials Science and Engineering, National Chiao Tung University, No. 1001, Ta-Hsueh Rd, Hsinchu, 30010, Taiwan.
Tsung-Ying
TY
Hsieh
Department of Materials Science and Engineering, National Chiao Tung University, No. 1001, Ta-Hsueh Rd, Hsinchu, 30010, Taiwan.
Wei-Chen
WC
Huang
Department of Materials Science and Engineering, National Chiao Tung University, No. 1001, Ta-Hsueh Rd, Hsinchu, 30010, Taiwan.
Yi-Da
YD
Kang
Department of Materials Science and Engineering, National Chiao Tung University, No. 1001, Ta-Hsueh Rd, Hsinchu, 30010, Taiwan.
Chao-Yi
CY
Chu
Taiwan Institute of Neuroscience, National Chengchi University, No. 64, Sec. 2, Zhinan Rd., Wenshan Dist, Taipei City, 116, Taiwan.
Wen-Lin
WL
Liao
Department of Biomedical Engineering, National Yang Ming University, No. 155, Sec. 2, Linong St, Taipei, Taiwan.
You-Yin
YY
Chen
Department of Materials Science and Engineering, National Chiao Tung University, No. 1001, Ta-Hsueh Rd, Hsinchu, 30010, Taiwan.
San-Yuan
SY
Chen
D000818:Animals; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006108:Graphite / Q000008:administration & dosage* / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry; D019636:Neurodegenerative Diseases / Q000188:drug therapy*; D009474:Neurons / Q000187:drug effects*; D009496:Neurotensin / Q000008:administration & dosage* / Q000737:chemistry; D010087:Oxides / Q000008:administration & dosage* / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011094:Polyethyleneimine / Q000737:chemistry; D051381:Rats; D019265:Spectroscopy, Near-Infrared / Q000379:methods; D014162:Transfection / Q000379:methods
D010087:Oxides; D009496:Neurotensin; D006108:Graphite; D011094:Polyethyleneimine
central nervous system; gene transfection; graphene oxide; laser photothermal treatment; nanoparticles