title: Facile Construction of Chloroquine Containing PLGA-Based pDNA Delivery System for Efficient Tumor and Pancreatitis Targeting in Vitro and in Vivo.
abstract: Chloroquine diphosphate (CQ) was ingeniously used to take place of phosphate salt in traditional calcium phosphate coprecipitation method for pDNA transfection. With multiple roles of CQ in the novel Ca-CQ-pDNA complex including pDNA compaction and assistance in lysosome escape, the transfection efficiency of the pDNA was significantly increased relative to the traditional method. CQ did not intercalate into the DNA double helix as free CQ did, which was probably ascribed to the prior mixing of the pDNA with high concentration of calcium chloride. In order to construct efficacious vector for in vivo gene delivery, Ca-CQ-pDNA-PLGA-NPs was designed and prepared. With entrapment efficiency, particle size and pDNA integrity as screening conditions, the optimal prescription was obtained and CaPi-pDNA-PLGA-NPs made with classic calcium phosphate coprecipitation method after optimization was also prepared as control to systematically study the role of CQ in the novel vector. Physical characters of the vectors were comprehensively studied using TEM, DSC, and XRD. The safety of the vector both in vitro and in vivo was evaluated using MTT, hemolysis test, and histological sections. The Ca-CQ-pDNA-PLGA-NPs dramatically enhanced the gene tranfection efficiency in Human Embryonic kidney HEK293 cells compared with the CaPi-pDNA-PLGA-NPs and presented an increasing gene transfection for up 144 h. The relative fast release of the CQ compared with pDNA from the nanoparticles was responsive for the increased transfection. The Did-labeled-Ca-CQ-pDNA-PLGA-NPs exhibited excellent tumor targeting efficiency and sustained circulation time in CT26 mouse model. The Ca-CQ-pDNA-PLGA-NP loaded with the plasmid pVITRO2 expressing mSurvivin-T34A protein gave 70% tumor inhibition rate, which was partially ascribed to CQ. The Ca-CQ-pDNA-PLGA-NPs showed high targeting efficiency in C57 acute pancreatitis model. In all, the Ca-CQ-pDNA-PLGA-NP was a promising candidate for targeted gene delivery to both tumor and pancreatitis.
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Chengli
authors.initials: C
authors.lastname: Yang
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.|#West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Hu
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.|#West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Cao
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Lijing
authors.initials: L
authors.lastname: Zhang
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.|#West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Pengxiang
authors.initials: P
authors.lastname: Zhou
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Gu
authors.initials: G
authors.lastname: He
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Xiangrong
authors.initials: X
authors.lastname: Song
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Aiping
authors.initials: A
authors.lastname: Tong
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Gang
authors.initials: G
authors.lastname: Guo
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.|‡Department of Gynecology, West China Second University Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: §Laboratory of Pharmaceutics, School of Medicine, Tsinghua University, Beijing 100084, P. R. China.
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Zhang
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Qian
authors.affiliation: ‡Department of Gynecology, West China Second University Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Qi
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.|∥Department of Cerebral Surgery, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Liangxue
authors.initials: L
authors.lastname: Zhou
authors.affiliation: †State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P. R. China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
mesh_terms: D000818:Animals; D002152:Calorimetry, Differential Scanning; D002738:Chloroquine / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry*; D008297:Male; D051379:Mice; D046529:Microscopy, Electron, Transmission; D010195:Pancreatitis / Q000628:therapy*; D010957:Plasmids / Q000008:administration & dosage*; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer
chemical_list: D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D002738:Chloroquine
keywords: Ca-CQ-pDNA-PLGA-NP; chloroquine containing PLGA-based pDNA delivery system; enhanced lysosome escape; pancreatitis targeting; tumor targeting