AIE-enabled transfection-free identification and isolation of viable cell subpopulations differing in the level of autophagy

Wenbin Zhang; Pengfei Wei; Liu Liu; Tao Ding; Yinyin Yang; Peipei Jin; Li Zhang; Zhibin Zhao; Meimei Wang; Bochuan Hu; Xin Jin; Zeng Xu; Han Zhang; Yang Song; Liansheng Wang; Suqin Zhong; Jing Chen; Zhenyu Yang; Ziying Chen; Yu Wu; Zhiming Ye; Youcui Xu; Yunjiao Zhang; Long-Ping Wen

doi:10.1080/15548627.2023.2235197

AIE-enabled transfection-free identification and isolation of viable cell subpopulations differing in the level of autophagy

Autophagy. 2023 Dec;19(12):3062-3078. doi: 10.1080/15548627.2023.2235197. Epub 2023 Aug 2.

Authors

Wenbin Zhang^{1

2}, Pengfei Wei³, Liu Liu⁴, Tao Ding⁵, Yinyin Yang¹, Peipei Jin⁵, Li Zhang⁶, Zhibin Zhao¹, Meimei Wang⁵, Bochuan Hu⁵, Xin Jin⁵, Zeng Xu⁷, Han Zhang⁷, Yang Song⁵, Liansheng Wang¹, Suqin Zhong¹, Jing Chen⁶, Zhenyu Yang⁸, Ziying Chen⁵, Yu Wu¹, Zhiming Ye¹, Youcui Xu¹, Yunjiao Zhang⁵, Long-Ping Wen¹

Affiliations

¹ Medical Research Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, Guangdong, China.
² State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, Guangdong, China.
³ School of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, Shandong, China.
⁴ Department of Clinical Laboratory, the First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
⁵ School of Medicine, School of Biomedical Sciences and Engineering, National Engineering Research Center for Tissue Restoration and Reconstruction and Key Laboratory of Biomedical Engineering of Guangdong Province, South China University of Technology, Guangzhou, Guangdong, China.
⁶ Department of Urology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
⁷ Center for Aggregation-Induced Emission, State Key Laboratory of Luminescent Materials and Devices, South China University of Technology, Guangzhou, Guangdong, China.
⁸ China-Singapore International Joint Research Institute, Guangzhou, Guangdong, China.

PMID: 37533292
PMCID: PMC10621245 (available on 2024-08-02)
DOI: 10.1080/15548627.2023.2235197

Abstract

3-MA, 3-methyladenine; AIE, aggregation-induced emission; AIEgens, aggregation-induced emission luminogens; ATG5, autophagy related 5; BMDM, bone marrow-derived macrophage; CQ, chloroquine; DiD, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine perchlorate; DiO, 3,3'-dioctadecyloxacarbocyanine perchlorate; DMSO, dimethyl sulfoxide; d-THP-1, differentiated THP-1; FACS, fluorescence activated cell sorting; FBS, fetal bovine serum; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; GABARAP, GABA type A receptor-associated protein; GFP, green fluorescent protein; HBSS, Hanks' balanced salt solution; HPLC, high-performance liquid chromatography; HRP, horseradish peroxidase; IL1B, interleukin 1 beta; KT, an AIE probe composed of a cell-penetrating peptide and an AIEgen tetraphenyl ethylene; LC3-II, lipidated LC3; LDH, lactate dehydrogenase; LIR, LC3-interacting region; LKR, engineered molecular probe composed of an LC3-interacting peptide, a cell-penetrating peptide and a non-AIE fluorescent molecule rhodamine; LKT, engineered molecular probe composed of an LC3-interacting peptide, a cell-penetrating peptide and an AIEgen tetraphenyl ethylene; LPS, lipopolysaccharide; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; MEF, mouse embryonic fibroblast; mRFP, monomeric red fluorescent protein; NHS, N-hydroxysuccinimide; NLRP3, NLR family pyrin domain containing 3; PBS, phosphate-buffered saline; PCC, pearson's correlation coefficient; PL, photoluminescence; PMA, phorbol 12-myristate 13-acetate; RAP, rapamycin; RIM, restriction of intramolecular motions; s.e.m., standard error of the mean; SPR, surface plasmon resonance; SQSTM1/p62, sequestosome 1; TAX1BP1, Tax1 binding protein 1; TPE, tetraphenylethylene; TPE-yne, 1-(4-ethynylphenyl)-1,2,2-triphenylethene; Tre, trehalose; u-THP-1: undifferentiated THP-1; UV-Vis, ultraviolet visible.

Keywords: Aggregation-induced emission; LC3-interacting; autophagy; cell sorting; flow cytometry; transfection-free.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis Regulatory Proteins / metabolism
Autophagy*
Cell-Penetrating Peptides*
Ethylenes
Fibroblasts / metabolism
Green Fluorescent Proteins
Mice
Molecular Probes

Substances

Cell-Penetrating Peptides
Apoptosis Regulatory Proteins
Green Fluorescent Proteins
Molecular Probes
Ethylenes

Grants and funding

This work was supported by grants from the National Natural Science Foundation of China (82150118, T2222014, 32171375, 91859103, 32071398, 82001671, 81601600), Guangdong Key R&D Project (2020B0101030006, 2020B1515120096), Taishan Scholars Program of Shandong Province (Tsqn202103112), Scientific research start-up fund of Binzhou Medical University (BY2021KYQD21), and Development Plan of Youth Innovation Team in Colleges and universities of Shandong Province (2021KJ052).