Engineered phage with cell-penetrating peptides for intracellular bacterial infections

作者全名："Zhao, Min; Tan, Xin; Liu, Zi-qiang; Dou, Lei; Liu, Dong; Pan, Yong-jun; Ma, Ying-fei; Yu, Jia-lin"

作者地址："[Zhao, Min; Yu, Jia-lin] Chongqing Med Univ, Dept Neonatol,Childrens Hosp, Natl Clin Res Ctr Child Hlth & Disorders,Chongqin, Minist Educ,Key Lab Child Dev & Disorders,Chongqi, Chongqing, Peoples R China; [Tan, Xin; Liu, Zi-qiang; Ma, Ying-fei] Chinese Acad Sci, CAS Key Lab Quantitat Engn Biol, Shenzhen Inst Synthet Biol, Shenzhen Inst Adv Technol, Shenzhen, Peoples R China; [Dou, Lei] Southern Univ Sci & Technol Hosp, Dept Neonatol, Shenzhen, Peoples R China; [Liu, Dong] Shenzhen Peoples Hosp, Dept Neonatol, Shenzhen, Peoples R China; [Pan, Yong-jun] Southern Univ Sci & Technol Hosp, Dept Crit Care Med, Shenzhen, Peoples R China"

通信作者："Yu, JL (通讯作者)，Chongqing Med Univ, Dept Neonatol,Childrens Hosp, Natl Clin Res Ctr Child Hlth & Disorders,Chongqin, Minist Educ,Key Lab Child Dev & Disorders,Chongqi, Chongqing, Peoples R China.; Tan, X; Ma, YF (通讯作者)，Chinese Acad Sci, CAS Key Lab Quantitat Engn Biol, Shenzhen Inst Synthet Biol, Shenzhen Inst Adv Technol, Shenzhen, Peoples R China."

来源：MSYSTEMS

ESI学科分类：MICROBIOLOGY

WOS号：WOS:001162804100037

JCR分区：Q1

影响因子：5

年份：2023

卷号：8

期号：5

开始页：　

结束页：　

文献类型：Article

关键词：phage engineering; cell-penetrating peptide; phage therapy; intracellular infection

摘要："Salmonella infection poses a critical challenge to global public health, and the situation is exacerbated by the increasing prevalence of antibiotic resistance. Bacteriophages (phages) are increasingly being used as antimicrobial agents due to their ability to kill specific bacteria. However, the low cellular uptake of phages has limited their use in treating intracellular bacterial infections. Here, we present a study using engineered phages with cell-penetrating peptides (CPPs) for enhancing the internalization efficiency of phages to inhibit bacterial intracellular infections. Through bioinformatic analysis, we identified a phage-encoded protein harboring an immunoglobulin-like (Ig-like) domain as the potential target for phage display. Using a CRISPR-Cas9-based method, we successfully displayed short peptides on GP94, an Ig-like domain-containing protein, of Salmonella phage selz. We improved phage intracellular uptake in multiple cell types by fusion of various CPPs to GP94. Notably, the phage selz(HA-TAT) showed promising results in enhancing the intracellular inhibition of Salmonella in different cells. Our research provides a straightforward strategy for displaying CPPs on non-model phages, offering a promising novel and effective therapeutic approach for treating intracellular and drug-resistant bacteria. IMPORTANCE Salmonella infection is a significant threat to global public health, and the increasing prevalence of antibiotic resistance exacerbates the situation. Therefore, finding new and effective ways to combat this pathogen is essential. Phages are natural predators of bacteria and can be used as an alternative to antibiotics to kill specific bacteria, including drug-resistant strains. One significant limitation of using phages as antimicrobial agents is their low cellular uptake, which limits their effectiveness against intracellular bacterial infections. Therefore, finding ways to enhance phage uptake is crucial. Our study provides a straightforward strategy for displaying cell-penetrating peptides on non-model phages, offering a promising novel and effective therapeutic approach for treating intracellular and drug-resistant bacteria. This approach has the potential to address the global challenge of antibiotic resistance and improve public health outcomes."

基金机构："Ministry of Science and Technology of China; National Key R&D Program of China [2018YFA0903100]; National Natural Science Foundation of China Fund Project [81971431, 32001038]; Strategic Priority Research Program of the Chinese Academy of Sciences [XDB29050500]; Shenzhen Institute of Synthetic Biology ScientificScientific Research Program [JCHZ20200001]; Shenzhen Science and Technology Innovation Free Exploration Project [JCYJ20220530112602005]; Shenzhen Outstanding Scientific Innovation Talents Training Project [RCBS20210706092214015]"

基金资助正文："This work was supported by the Ministry of Science and Technology of China, National Key R&D Program of China (2018YFA0903100), the National Natural Science Foundation of China Fund Project (no. 81971431 and no. 32001038), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29050500), Shenzhen Institute of Synthetic Biology ScientificScientific Research Program (JCHZ20200001), Shenzhen Science and Technology Innovation Free Exploration Project (no. JCYJ20220530112602005), and Shenzhen Outstanding Scientific Innovation Talents Training Project (no. RCBS20210706092214015)."