Mechanisms of transcellular transport of wheat germ agglutinin-functionalized polymeric nanoparticles in Caco-2 cells.
Transcellular transport is essential for transmucosal and plasma-to-tissue drug delivery by nanoparticles, whereas its fundamental pathways have not been fully clarified. In this study, an in-depth investigation was conducted into the intracellular itinerary and the transcytosis pathway of wheat germ agglutinin-functionalized nanoparticles (WGA-NP) with various polymer architectures in the Caco-2 cell model. GFP-Rabs, Rab4, Rab5, Rab7, Rab11, GTPases served as key regulators of vesicular transport, and their mutants were transfected to Caco-2 cells respectively to determine the cellular itinerary of WGA-NP and the role of Rabs therein. Transcytosis inhibition experiments indicated that transcellular transport of WGA-NP (PEG(3000)-PLA(40000) formulation) happened in a cytoskeleton-dependent manner and majorly by means of clathrin-mediated mechanism. Intracellular transport, especially the endolysosome pathway was found largely contribute to the transcytosis of WGA-NP. WGA-NP with shorter surface PEG length (2000) resulted in higher cellular association and more colocalization with the clathrin-mediated transport pathway, while that with longer surface PEG length (5000) avoided the clathrin-mediated transport pathway but achieved higher transcytosis after 4 h incubation. WGA-NP with PLGA as the core materials obtained elevated lysosome escape and enhanced transcytosis after 2 h incubation. These findings provided important evidence for the role of polymer architectures in modulating cellular transport of functionalized nanocarriers, and would be helpful in improving carrier design to enhance drug delivery.
Department of Pharmacology, Institute of Medical Sciences, Shanghai Jiaotong University School of Medicine, 280 South Chongqing Road, Shanghai 200025, PR China.
Qingxiang
Q
Song
Lei
L
Yao
Meng
M
Huang
Quanyin
Q
Hu
Qin
Q
Lu
Bingxian
B
Wu
Hong
H
Qi
Zhengxing
Z
Rong
Xinguo
X
Jiang
Xiaoling
X
Gao
Jun
J
Chen
Hongzhuan
H
Chen
D018938:Caco-2 Cells; D003374:Coumarins / Q000737:chemistry / Q000493:pharmacokinetics; D004337:Drug Carriers / Q000737:chemistry / Q000493:pharmacokinetics*; D005822:Genetic Vectors; D006801:Humans; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry / Q000493:pharmacokinetics; D011098:Polyglactin 910 / Q000737:chemistry / Q000493:pharmacokinetics; D057900:Transcytosis / Q000187:drug effects* / Q000502:physiology; D014909:Wheat Germ Agglutinins / Q000737:chemistry / Q000493:pharmacokinetics*; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
D003374:Coumarins; D004337:Drug Carriers; D050505:Mutant Proteins; D014909:Wheat Germ Agglutinins; C542623:monomethoxypolyethyleneglycol-polylactide block copolymer; C102418:poly(lactic-glycolic acid)-poly(ethyleneglycol) copolymer; D011098:Polyglactin 910; D011092:Polyethylene Glycols; D020691:rab GTP-Binding Proteins