In situ repurposing of dendritic cells with CRISPR/Cas9-based nanomedicine to induce transplant tolerance

Yue Zhang; Song Shen; Gui Zhao; Cong-Fei Xu; Hou-Bing Zhang; Ying-Li Luo; Zhi-Ting Cao; Jia Shi; Zhi-Bin Zhao; Zhe-Xiong Lian; Jun Wang

doi:10.1016/j.biomaterials.2019.119302

In situ repurposing of dendritic cells with CRISPR/Cas9-based nanomedicine to induce transplant tolerance

Biomaterials. 2019 Oct:217:119302. doi: 10.1016/j.biomaterials.2019.119302. Epub 2019 Jun 24.

Authors

Yue Zhang¹, Song Shen², Gui Zhao¹, Cong-Fei Xu³, Hou-Bing Zhang⁴, Ying-Li Luo⁵, Zhi-Ting Cao⁶, Jia Shi⁶, Zhi-Bin Zhao⁷, Zhe-Xiong Lian⁷, Jun Wang⁸

Affiliations

¹ Guangzhou Digestive Disease Center, Guangzhou First People's Hospital, School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, 510006, PR China; Hefei National Laboratory for Physical Sciences at the Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, 230027, PR China.
² Chronic Disease Laboratory, Institutes for Life Sciences and School of Medicine, South China University of Technology, Guangzhou, 510006, PR China; National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, Guangzhou, 510006, PR China.
³ Guangzhou Digestive Disease Center, Guangzhou First People's Hospital, School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, 510006, PR China; Chronic Disease Laboratory, Institutes for Life Sciences and School of Medicine, South China University of Technology, Guangzhou, 510006, PR China. Electronic address: xucf@scut.edu.cn.
⁴ Hefei National Laboratory for Physical Sciences at the Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, 230027, PR China.
⁵ Guangzhou Digestive Disease Center, Guangzhou First People's Hospital, School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, 510006, PR China; Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou, 510006, PR China.
⁶ Guangzhou Digestive Disease Center, Guangzhou First People's Hospital, School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, 510006, PR China.
⁷ Chronic Disease Laboratory, Institutes for Life Sciences and School of Medicine, South China University of Technology, Guangzhou, 510006, PR China.
⁸ Guangzhou Digestive Disease Center, Guangzhou First People's Hospital, School of Biomedical Sciences and Engineering, Guangzhou International Campus, South China University of Technology, Guangzhou, 510006, PR China; National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, Guangzhou, 510006, PR China; Key Laboratory of Biomedical Engineering of Guangdong Province, and Innovation Center for Tissue Restoration and Reconstruction, South China University of Technology, Guangzhou, 510006, PR China; Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou, 510005, PR China. Electronic address: mcjwang@scut.edu.cn.

PMID: 31271858
DOI: 10.1016/j.biomaterials.2019.119302

Abstract

Organ transplantation is the only effective method to treat end-stage organ failure. However, it is continuously plagued by immune rejection, which is mostly caused by T cell-mediated reactions. Dendritic cells (DCs) are professional antigen-presenting cells, and blocking the costimulatory signaling molecule CD40 in DCs inhibits T cell activation and induces transplant tolerance. In this study, to relieve graft rejection, Cas9 mRNA (mCas9) and a guide RNA targeting the costimulatory molecule CD40 (gCD40) were prepared and encapsulated into poly(ethylene glycol)-block-poly(lactide-co-glycolide) (PEG-b-PLGA)-based cationic lipid-assisted nanoparticles (CLAN), denoted CLAN_mCas9/gCD40. CLAN effectively delivered mCas9/gCD40 into DCs and disrupted CD40 in DCs at the genomic level both in vitro and in vivo. After intravenous injection into an acute mouse skin transplant model, CLAN_mCas9/gCD40-mediated CD40 disruption significantly inhibited T cell activation, which reduced graft damage and prolonged graft survival. This work provides a promising strategy for reprogramming DCs with nanoparticles carrying the CRISPR/Cas9 system to abate transplant rejection.

Keywords: CRISPR/Cas9; Dendritic cell; Gene editing; Nanoparticle; Transplant tolerance.

Publication types

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

MeSH terms

Animals
CD40 Antigens / metabolism
CRISPR-Associated Protein 9 / metabolism*
CRISPR-Cas Systems / genetics*
Dendritic Cells / immunology*
Disease Models, Animal
Endocytosis
Graft Rejection / immunology
Graft Survival / immunology
Lymphocyte Activation / immunology
Mice, Inbred BALB C
Mice, Inbred C57BL
Nanomedicine*
RNA, Guide, CRISPR-Cas Systems / metabolism
Transplantation Tolerance / immunology*

Substances

CD40 Antigens
RNA, Guide, CRISPR-Cas Systems
CRISPR-Associated Protein 9