title: Killing colon cancer cells through PCD pathways by a novel hyaluronic acid-modified shell-core nanoparticle loaded with RIP3 in combination with chloroquine.
abstract: Due to extensive apoptosis defects and multidrug resistance, there is great interest regarding non-apoptotic programmed cell death (PCD) pathways, such as lysosomal-mediated programmed cell death (LM-PCD), necroptosis and autophagy. Because there is an intricate effector network among these PCD pathways, it is expected that they may act synergistically in cancer therapy. In this study, chloroquine (CQ) was found to significantly upregulate receptor-interacting protein kinase 3 (RIP3) expression, and RIP3 were involved in CQ-related autophagy. Overexpressed-eGFP-RIP3 co-localized with the selective autophagy receptor p62. mRIP3 overexpression in combination with CQ markedly increased the inhibition rate relative to that observed in the CQ-treatment group. Several experiments, including Hoechst staining, transmission electron microscopy (TEM) observation, the high-mobility group box 1 (HMGB1) release assay, Annexin V/PI staining and immunoblotting of proteins included in PCD pathways, verified that mRIP3 overexpression in combination with CQ induced lysosomal membrane permeabilization (LMP) and necroptosis of cancer cells, leading to cancer cell death. For tumor-targeted delivery, hyaluronic acid (HA)-modified, lipid-coated PLGA nanoparticles loaded with mRIP3-pDNA were prepared and characterized using a particle sizer, differential scanning calorimetry (DSC) and TEM. The nanoparticles exhibited ideal biocompatibility and good tumor-targeting efficiency, and the tumor inhibition rate of HA-Lip-PEI-mRIP3-PLGA-NPs + CQ was 80.2% in the CT26 mouse model. In this study, we attempted to treat tumors by inducing several alternative PCD pathways to shed light on the combination therapy of alternative PCD inducers.
authors.affiliation: State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR China.
authors.forename: Xueyan
authors.initials: X
authors.lastname: Hou
authors.affiliation: Department of Clinical Pharmacy, School of Pharmacy, Zunyi Medical University, 6#, Xuefu Xi Road, Zunyi, Guizhou, 563006, PR China.
authors.forename: Chengli
authors.initials: C
authors.lastname: Yang
authors.affiliation: Medical Research Center, The First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Zhengzhou, Henan, 450052, PR 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, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Hu
authors.affiliation: College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610041, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR China.
authors.forename: Hua
authors.initials: H
authors.lastname: Cao
authors.affiliation: Department of Gynecology, West China Second University Hospital, Sichuan University, Chengdu, 610041, PR China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR 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, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR China.
authors.forename: Changyang
authors.initials: C
authors.lastname: Gong
authors.affiliation: Laboratory of Pharmaceutics, School of Medicine, Tsinghua University, 30#, Shuangqing Road, Haidian Dist, Beijing, 100084, PR 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, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR 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, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.affiliation: State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, 17#, Section 3, Ren Min Nan Road, Chengdu, Sichuan, 610041, PR China. Electronic address: zhengyu82@scu.edu.cn.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D002738:Chloroquine / Q000008:administration & dosage*; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D006820:Hyaluronic Acid / Q000737:chemistry*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053769:Nanocapsules / Q000008:administration & dosage / Q000737:chemistry* / Q000648:ultrastructure; D058726:Nanoconjugates / Q000008:administration & dosage / Q000737:chemistry / Q000648:ultrastructure; D058608:Nanopores / Q000648:ultrastructure; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000008:administration & dosage / Q000493:pharmacokinetics*; D016896:Treatment Outcome
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D053769:Nanocapsules; D058726:Nanoconjugates; D002738:Chloroquine; D006820:Hyaluronic Acid; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C485615:Ripk3 protein, mouse
keywords: Autophagy; Chloroquine; Hyaluronic acid-modified shell-core nanoparticle; Lysosomal membrane permeabilization (LMP); Necroptosis; mRIP3