Targeting of NLRP3 inflammasome with gene editing for the amelioration of inflammatory diseases

Congfei Xu; Zidong Lu; Yingli Luo; Yang Liu; Zhiting Cao; Song Shen; Hongjun Li; Jing Liu; Kaige Chen; Zhiyao Chen; Xianzhu Yang; Zhen Gu; Jun Wang

doi:10.1038/s41467-018-06522-5

Targeting of NLRP3 inflammasome with gene editing for the amelioration of inflammatory diseases

Nat Commun. 2018 Oct 5;9(1):4092. doi: 10.1038/s41467-018-06522-5.

Authors

Congfei Xu^{1

2

3}, Zidong Lu^{1

2}, Yingli Luo⁴, Yang Liu⁴, Zhiting Cao⁴, Song Shen^{1

2

3}, Hongjun Li^{1

2

3}, Jing Liu⁴, Kaige Chen⁴, Zhiyao Chen⁴, Xianzhu Yang^{1

2

3}, Zhen Gu⁵, Jun Wang^{6

7

8

9}

Affiliations

¹ Guangzhou First People's Hospital, School of Medicine, South China University of Technology, 510006, Guangzhou, Guangdong, China.
² National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, 510006, Guangzhou, Guangdong, China.
³ Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, School of Biomedical Science and Engineering, South China University of Technology, Guangzhou International Campus, 510006, Guangzhou, Guangdong, China.
⁴ School of Life Sciences, University of Science and Technology of China, 230027, Hefei, Anhui, China.
⁵ Department of Bioengineering, California Nanosystems Institute, University of California, Los Angeles, CA, 90095, USA.
⁶ Guangzhou First People's Hospital, School of Medicine, South China University of Technology, 510006, Guangzhou, Guangdong, China. mcjwang@scut.edu.cn.
⁷ National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, 510006, Guangzhou, Guangdong, China. mcjwang@scut.edu.cn.
⁸ Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, School of Biomedical Science and Engineering, South China University of Technology, Guangzhou International Campus, 510006, Guangzhou, Guangdong, China. mcjwang@scut.edu.cn.
⁹ Key Laboratory of Biomedical Engineering of Guangdong Province, South China University of Technology, 510006, Guangzhou, Guangdong, China. mcjwang@scut.edu.cn.

Abstract

The NLRP3 inflammasome is a well-studied target for the treatment of multiple inflammatory diseases, but how to promote the current therapeutics remains a large challenge. CRISPR/Cas9, as a gene editing tool, allows for direct ablation of NLRP3 at the genomic level. In this study, we screen an optimized cationic lipid-assisted nanoparticle (CLAN) to deliver Cas9 mRNA (mCas9) and guide RNA (gRNA) into macrophages. By using CLAN encapsulating mCas9 and gRNA-targeting NLRP3 (gNLRP3) (CLAN_mCas9/gNLRP3), we disrupt NLRP3 of macrophages, inhibiting the activation of the NLRP3 inflammasome in response to diverse stimuli. After intravenous injection, CLAN_mCas9/gNLRP3 mitigates acute inflammation of LPS-induced septic shock and monosodium urate crystal (MSU)-induced peritonitis. In addition, CLAN_mCas9/gNLRP3 treatment improves insulin sensitivity and reduces adipose inflammation of high-fat-diet (HFD)-induced type 2 diabetes (T2D). Thus, our study provides a promising strategy for treating NLRP3-dependent inflammatory diseases and provides a carrier for delivering CRISPR/Cas9 into macrophages.

Publication types

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

MeSH terms

Amino Acid Sequence
Animals
Apoptosis Regulatory Proteins / metabolism
CRISPR-Cas Systems
Calcium-Binding Proteins / metabolism
Caspase Activation and Recruitment Domain
Female
Gene Editing*
Genetic Therapy*
Inflammasomes / genetics*
Inflammation / therapy*
Male
Mice, Inbred C57BL
NLR Family, Pyrin Domain-Containing 3 Protein / genetics*
NLR Family, Pyrin Domain-Containing 3 Protein / metabolism
Peritonitis / immunology
Protein Conformation

Substances

Apoptosis Regulatory Proteins
Calcium-Binding Proteins
Inflammasomes
Ipaf protein, mouse
NLR Family, Pyrin Domain-Containing 3 Protein
Nlrp3 protein, mouse