title: Cationic nanocarriers induce cell necrosis through impairment of Na(+)/K(+)-ATPase and cause subsequent inflammatory response.
abstract: Nanocarriers with positive surface charges are known for their toxicity which has limited their clinical applications. The mechanism underlying their toxicity, such as the induction of inflammatory response, remains largely unknown. In the present study we found that injection of cationic nanocarriers, including cationic liposomes, PEI, and chitosan, led to the rapid appearance of necrotic cells. Cell necrosis induced by cationic nanocarriers is dependent on their positive surface charges, but does not require RIP1 and Mlkl. Instead, intracellular Na(+) overload was found to accompany the cell death. Depletion of Na(+) in culture medium or pretreatment of cells with the Na(+)/K(+)-ATPase cation-binding site inhibitor ouabain, protected cells from cell necrosis. Moreover, treatment with cationic nanocarriers inhibited Na(+)/K(+)-ATPase activity both in vitro and in vivo. The computational simulation showed that cationic carriers could interact with cation-binding site of Na(+)/K(+)-ATPase. Mice pretreated with a small dose of ouabain showed improved survival after injection of a lethal dose of cationic nanocarriers. Further analyses suggest that cell necrosis induced by cationic nanocarriers and the resulting leakage of mitochondrial DNA could trigger severe inflammation in vivo, which is mediated by a pathway involving TLR9 and MyD88 signaling. Taken together, our results reveal a novel mechanism whereby cationic nanocarriers induce acute cell necrosis through the interaction with Na(+)/K(+)-ATPase, with the subsequent exposure of mitochondrial damage-associated molecular patterns as a key event that mediates the inflammatory responses. Our study has important implications for evaluating the biocompatibility of nanocarriers and designing better and safer ones for drug delivery.
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Xiawei
authors.initials: X
authors.lastname: Wei
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Shao
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Zhiyao
authors.initials: Z
authors.lastname: He
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Tinghong
authors.initials: T
authors.lastname: Ye
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Min
authors.initials: M
authors.lastname: Luo
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Yaxiong
authors.initials: Y
authors.lastname: Sang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Shuntao
authors.initials: S
authors.lastname: Luo
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Shengyong
authors.initials: S
authors.lastname: Yang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Changyang
authors.initials: C
authors.lastname: Gong
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Maling
authors.initials: M
authors.lastname: Gou
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Hongxing
authors.initials: H
authors.lastname: Deng
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Yinglan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Hanshuo
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Senyi
authors.initials: S
authors.lastname: Deng
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Chengjian
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Qian
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Jiong
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China.
authors.forename: Jiahuai
authors.initials: J
authors.lastname: Han
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Department of Biochemistry and Molecular Biology, Peking Union Medical College, Beijing 100005, China.
authors.forename: Chengyu
authors.initials: C
authors.lastname: Jiang
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Medicine and Health Sciences, University of North Dakota, Grand Forks, ND 58202, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Wu
authors.affiliation: Key Laboratory of Drug Targeting of Ministry of Education, State Key Laboratory of Biotherapy/Collaborative Innovation Center, West China School of Pharmacy, Sichuan University, No. 17, Block 3, Southern Renmin Road, Chengdu, Sichuan 610041, China.
authors.forename: Zhirong
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002412:Cations / Q000737:chemistry; D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry / Q000633:toxicity; D004272:DNA, Mitochondrial / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry / Q000633:toxicity*; D006801:Humans; D007249:Inflammation / Q000473:pathology; D008081:Liposomes / Q000737:chemistry / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry / Q000633:toxicity*; D009336:Necrosis; D010042:Ouabain / Q000494:pharmacology; D011094:Polyethyleneimine / Q000737:chemistry / Q000633:toxicity; D015398:Signal Transduction / Q000187:drug effects; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051217:Toll-Like Receptor 9 / Q000378:metabolism
chemical_list: D002412:Cations; D004272:DNA, Mitochondrial; D004337:Drug Carriers; D008081:Liposomes; D053594:Myeloid Differentiation Factor 88; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9; D010042:Ouabain; D011094:Polyethyleneimine; D048271:Chitosan; D000254:Sodium-Potassium-Exchanging ATPase