Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity

Le-Yang Wu; Jia-Hui Qiu; Xin-Yue Qiao; Lin Li; Li-Yuan Qiao; Chen-Yang Li; Ying Sun; Shu-Hui Zhang; Zeng-Zheng Du; Xiao-Yao Chang; Cheng Cheng; Bo-Hao Wang; Yi-Han Xiao; Lin Lin; Zi-Chun Hua

doi:10.1016/j.mmr.2026.100030

Your Location：

Home >

Browse articles >

Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity

RESEARCH | Updated：2026-05-11

- Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity
- Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity
- 军事医学研究（英文） 2026年
- Affiliations：
  
  State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China
  Changzhou High-Tech Research Institute of Nanjing University and Jiangsu TargetPharma Laboratories, Inc., Changzhou 213164, Jiangsu, China
  School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211816, China
  School of Food and Biological Engineering, Jiangsu University, Nanjing 212013, China
  Faculty of Pharmaceutical Sciences, Xinxiang Medical University, Xinxiang 453002, Henan, China
- Author bio：
  
  *Le-Yang Wu, lywu@nju.edu.cn;
  Lin Lin, linl@ujs.edu.cn;
  Zi-Chun Hua, zchua@nju.edu.cn
- Funds：
  
  the National Natural Science Foundation of China(82303774;82130106);the Natural Science Foundation of Jiangsu Province(BK20243001;BG2024026;BK20230165;BE2023695;BY20241051);the Changzhou Municipal Department of Science and Technology(CE20246001;CJ20250023)
- DOI：10.1016/j.mmr.2026.100030
  中图分类号：
- 收稿：2025-06-21，
  
  录用：2026-04-03，
  
  纸质出版：2026-04
- Accepted：
Scan QR Code
Le-Yang Wu, Jia-Hui Qiu, Xin-Yue Qiao, 等. Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity[J/OL]. 军事医学研究（英文）, 2026.

Wu LY, Qiu JH, Qiao XY, Li L, Qiao LY, Li CY, et al. Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity. Mil Med Res. 2026;13(1):100030.
Le-Yang Wu, Jia-Hui Qiu, Xin-Yue Qiao, 等. Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity[J/OL]. 军事医学研究（英文）, 2026. DOI： 10.1016/j.mmr.2026.100030.

Wu LY, Qiu JH, Qiao XY, Li L, Qiao LY, Li CY, et al. Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity. Mil Med Res. 2026;13(1):100030. DOI： 10.1016/j.mmr.2026.100030.

摘要

Abstract

Background:

Bacteria-mediated cancer therapy leverages bacteria to modulate the tumor immune microenvironment and deliver therapeutics. However

its clinical application is limited by toxicity

off-target effects

and uncontrolled drug release. Improving tumor targeting and precise payload delivery through rational bacterial engineering is essential for increasing efficacy and safety.

Methods:

An attenuated

Salmonella ΔhtrA::luxI

-VNP20009 strain expressing OmpA-SpyTag (AISI-ST) was constructed for the modular surface conjugation of SpyCatcherΔ (SC)-fused quadruple arginine-glycine-aspartic acid (RGD) peptides (named AISI-ST/SC-RGD×4) and for building biointerfaces for enhanced tumor adhesion via RGD-mediated integrin αvβ3 interactions. The tumor-bearing mice received intravenous injections of AISI-ST/SC-RGD×4

and their biodistribution was analyzed using bioluminescence imaging and colony-forming unit (CFU) counts. Quorum-sensing (QS)-regulated high-temperature requirement A (HtrA) and anti-programmed cell death protein 1 (anti-PD1) nanobody expression based on the

LuxI

promoter in strains was validated by Western blotting. Immune responses were assessed using flow cytometry.

Results:

The incubation of the fused proteins with the AISI-ST strain for 1 h was sufficient to form a stable biological

interface. The quadruple RGD-modified bacteria (AISI-ST/SC-RGD×4) exhibited greater enrichment in various solid tumors and lung metastases with reduced off-target accumulation. QS induced the expression of the HtrA protein within tumors

resulting in enhanced extracellular polysaccharide-mediated immunogenicity to activate immune cells. Further expression of anti-PD1 nanobodies synergistically enhanced antitumor immunity

increasing the percentage of M1 macrophages (MACS) and CD8

T cell proliferation while suppressing M2 MACS and regulatory T cells (Tregs). This approach achieves potent tumor suppression via targeted immune remodeling.

Conclusions:

This study presents octopus-inspired engineered bacteria with a “plug-and-display” system and tumor-specific drug delivery that achieves enhanced tumor targeting and potent antitumor effects. This study describes a promising strategy for the precise and safe clinical translation of bacteria-mediated cancer immunotherapy.

关键词

Keywords

references

Dawson WR , Ebbell B . The papyrus ebers; the greatest egyptian medical document . J Egypt Archaeol . 1938 ; 24 : 250 .

Wu L , Bao F , Li L , Yin X , Hua Z . Bacterially mediated drug delivery and therapeutics: strategies and advancements . Adv Drug Deliv Rev . 2022 ; 187 : 114363 .

Kwon SY , Thi-Thu Ngo H , Son J , Hong Y , Min JJ . Exploiting bacteria for cancer immunotherapy . Nat Rev Clin Oncol . 2024 ; 21 ( 8 ): 569 - 89 .

Li L , Wu L , Qiao L , Zhang S , Luan X , Qiu J , et al . Precision tumor immunotherapy via a dual-gated macrophage-bacterial activation platform . Trends Biotechnol . 2026 ; 44 ( 2 ): 450 - 76 .

Wu L , Li C , Qiao L , Li L , Zhang S , Wang B , et al . Controlled pyroptosis of engineered macrophages enables biphasic anti-tumor via the release of oncolytic bacteria and inflammatory signals . Cell Rep . 2026 ; 45 ( 2 ): 116918 .

Wu L , Qiao L , Zhang S , Qiu J , Du Z , Sun Y , et al . Dual-engineered macrophage-microbe encapsulation for metastasis immunotherapy . Adv Mater . 2024 ; 36 ( 36 ): 2406140 .

Li F , Yang Z , Savage TM , Vincent RL , De Los Santos-Alexis K , Ahn A , et al . Programmable bacteria synergize with PD-1 blockade to overcome cancer cell-intrinsic immune resistance mechanisms . Sci Immunol . 2024 ; 9 ( 100 ): eadn9879 .

Sun D , Li Y , Yin X , Fan Y , Liu J , Wang Y , et al . Utilizing engineered bacteria as "cell factories" in vivo for intracellular RNA-loaded outer membrane vesicles' self-assembly in tumor treatment . ACS Nano . 2024 ; 18 ( 52 ): 35296 - 309 .

Wu L , Li L , Li S , Liu L , Xin W , Li C , et al . Macrophage-mediated tumor-targeted delivery of engineered Salmonella typhimurium VNP20009 in anti-PD1 therapy against melanoma . Acta Pharm Sin B . 2022 ; 12 ( 10 ): 3952 - 71 .

Wang L , Cao Z , Zhang M , Lin S , Liu J . Spatiotemporally controllable distribution of combination therapeutics in solid tumors by dually modified bacteria . Adv Mater . 2022 ; 34 ( 1 ): e2106669 .

Din MO , Danino T , Prindle A , Skalak M , Selimkhanov J , Allen K , et al . Synchronized cycles of bacterial lysis for in vivo delivery . Nature . 2016 ; 536 ( 7614 ): 81 - 5 .

Yelin I , Flett KB , Merakou C , Mehrotra P , Stam J , Snesrud E , et al . Genomic and epidemiological evidence of bacterial transmission from probiotic capsule to blood in ICU patients . Nat Med . 2019 ; 25 ( 11 ): 1728 - 32 .

Toso JF , Gill VJ , Hwu P , Marincola FM , Restifo NP , Schwartzentruber DJ , et al . Phase I study of the intravenous administration of attenuated Salmonella typhimurium to patients with metastatic melanoma . J Clin Oncol . 2002 ; 20 ( 1 ): 142 - 52 .

Heimann DM , Rosenberg SA . Continuous intravenous administration of live genetically modified Salmonella typhimurium in patients with metast atic melanoma . J Immunother . 2003 ; 26 ( 2 ): 179 - 80 .

Kolaczkowska E , Jenne CN , Surewaard BG , Thanabalasuriar A , Lee WY , Sanz MJ , et al . Molecular mechanisms of NET formation and degradation revealed by intravital imaging in the liver vasculature . Nat Commun . 2015 ; 6 : 6673 .

Zhang Y , Huo F , Cao Q , Jia R , Huang Q , Wang ZA , et al . FimH confers mannose-targeting ability to Bacillus Calmette-Guerin for improved immunotherapy in bladder cancer . J Immunother Cancer . 2022 ; 10 ( 3 ): e003939 .

Massa PE , Paniccia A , Monegal A , De Marco A , Rescigno M . Salmonella engineered to express CD20-targeting antibodies and a drug-converting enzyme can eradicate human lymphomas . Blood . 2013 ; 122 ( 5 ): 705 - 14 .

Xu X , Ding Y , Dong Y , Yuan H , Xia P , Qu C , et al . Nanobody-engineered biohybrid bacteria targeting gastrointestinal cancers induce robust STING-mediated anti-tumor immunity . Adv Sci (Weinh) . 2024 ; 11 ( 31 ): e2401905 .

Lång H . Outer membrane proteins as surface display systems . Int J Med Microbiol . 2000 ; 290 ( 7 ): 579 - 85 .

Park SH , Zheng JH , Vu Hong N , Jiang SN , Kim DY , Szardenings M , et al . RGD peptide cell-surface display enhances the targeting and therapeutic efficacy of attenuated Salmonella -mediated Cancer Therapy . Theranostics . 2016 ; 6 ( 10 ): 1672 - 82 .

Chen QW , Zhang Y , Bao P , Zhang XZ . Sialidase-chimeric bioengineered bacteria for tumor-sialoglycan-triggered solid tumor therapy . Nano Lett . 2024 ; 24 ( 33 ): 10362 - 71 .

Geng Z , Cao Z , Liu R , Liu K , Liu J , Tan W . Aptamer-assisted tumor localization of bacteria for enhanced biotherapy . Nat Commun . 2021 ; 12 ( 1 ): 6584 .

Xiao Y , Pan T , Da W , Liu Y , Chen S , Chen D , et al . Aptamer-drug conjugates-loaded bacteria for pancreatic cancer synergistic therapy . Signal Transduct Target Ther . 2024 ; 9 ( 1 ): 272 .

Cong Z , Tang S , Xie L , Yang M , Li Y , Lu D , et al . Magnetic-powered janus cell robots loaded with oncolytic adenovirus for active and targeted virotherapy of bladder cancer . Adv Mater . 2022 ; 34 ( 26 ): e2201042 .

Beygmoradi A , Homaei A , Hemmati R , Fernandes P . Recombinant protein expression: challenges in production and folding-related matters . Int J Biol Macromol . 2023 ; 233 : 123407 .

Banahene N , Kavunja HW , Swarts BM . Chemical reporters for bacterial glycans: development and applications . Chem Rev . 2022 ; 122 ( 3 ): 3336 - 413 .

Zakeri B , Fierer JO , Celik E , Chittock EC , Schwarz-Linek U , Moy VT , et al . Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin . Proc Natl Acad Sci U S A . 2012 ; 109 ( 12 ): e690 - 7 .

Sutherland AR , Alam MK , Geyer CR . Post-translational assembly of protein parts into complex devices by using SpyTag/SpyCatcher protein ligase . Chembiochem . 2019 ; 20 ( 3 ): 319 - 28 .

Kasaraneni N , Chamoun-Emanuelli AM , Wright G , Chen Z . Retargeting lentiviruses via SpyCatcher-SpyTag chemistry for gene delivery into specific cell types . mBio . 2017 ; 8 ( 6 ): e01860 - 17 .

Yan Y , Yang S , Mach KE , Chen G , Degraff DJ , Liao JC , et al . Microbial product cocktails for personalized cancer immunotherapy . Nat Commun . 2025 ; 16 ( 1 ): 10625 .

Prendergast BJ , Onishi KG , Zucker I . Female mice liberated for inclusion in neuroscience and biomedical research . Neurosci Biobehav Rev . 2014 ; 40 : 1 - 5 .

Wu L , Li L , Qiao L , Li C , Zhang S , Yin X , et al . Programmable bacteria with dynamic virulence modulation system for precision antitumor immunity . Adv Sci (Weinh) . 2024 ; 11 ( 36 ): e2404069 .

O'connell KE , Mikkola AM , Stepanek AM , Vernet A , Hall CD , Sun CC , et al . Practical murine hematopathology: a comparative review and implications for research . Comp Med . 2015 ; 65 ( 2 ): 96 - 113 .

Pina A , Kadri M , Arosio D , Dal Corso A , Coll JL , Gennari C , et al . Multimeric presentation of RGD peptidomimetics enhances integrin binding and tumor cell uptake . Chemistry . 2020 ; 26 ( 33 ): 7492 - 6 .

Dayan A , Fleminger G , Ashur-Fabian O . RGD-modified dihydrolipoamide dehydrogenase conjugated to titanium dioxide nanoparticles-switchable integrin-targeted photodynamic treatment of melanoma cells . RSC Adv . 2018 ; 8 ( 17 ): 9112 - 9 .

Benito-Jardón M , Strohmeyer N , Ortega-Sanchís S , Bharadwaj M , Moser M , Müller DJ , et al . αv-class integrin binding to fibronectin is solely mediated by RGD and unaffected by an RGE mutation . J Cell Biol . 2020 ; 219 ( 12 ): e202004198 .

Swofford CA , Van Dessel N , Forbes NS . Quorum-sensing Salmonella selectively trigger protein expression within tumors . Proc Natl Acad Sci U S A . 2015 ; 112 ( 11 ): 3457 - 62 .

Wu L , Li L , Yin X , Li C , Xin W , Liu L , et al . A SARS-CoV-2 oral vaccine development strategy based on the attenuated Salmonella type Ⅲ secretion system . J Appl Microbiol . 2022 ; 133 ( 4 ): 2484 - 500 .

Zhang X , Qi C , Guo Y , Zhou W , Zhang Y . Toll-like receptor 4-related immunostimulatory polysaccharides: primary structure, activity relationships, and possible interaction models . Carbohydr Polym . 2016 ; 149 : 186 - 206 .

Huang AC , Postow MA , Orlowski RJ , Mick R , Bengsch B , Manne S , et al . T-cell invigoration to tumour burden ratio associated with anti-PD-1 response . Nature . 2017 ; 545 ( 7652 ): 60 - 5 .

Walvekar P , Kumar P , Choonara YE . Long-acting vaccine delivery systems . Adv Drug Deliv Rev . 2023 ; 198 : 114897 .

Kung YJ , Lam B , Tseng SH , Macdonald A , Tu HF , Wang S , et al . Localization of Salmonella and albumin-IL-2 to the tumor microenvironment augments anticancer T cell immunity . J Biomed Sci . 2022 ; 29 ( 1 ): 57 .

Windemuth S , Hahn J , You J , Wang Z , Ding S , Tarrab S , et al . Iron-tannin coating reduces clearance and increases tumor colonization of systemically delivered bacteria . ACS Synth Biol . 2024 ; 13 ( 12 ): 3948 - 60 .

Wu L , Du Z , Li L , Qiao L , Zhang S , Yin X , et al . Camouflaging attenuated Salmonella by cryo-shocked macrophages for tumor-targeted therapy . Signal Transduct Target Ther . 2024 ; 9 ( 1 ): 14 .

Litchfield K , Reading JL , Puttick C , Thakkar K , Abbosh C , Bentham R , et al . Meta-analysis of tumor- and T cell-intrinsic mechanisms of sensitization to checkpoint inhibition . Cell . 2021 ; 184 ( 3 ): 596 - 614.e14 .

Gurbatri CR , Lia I , Vincent R , Coker C , Castro S , Treuting PM , et al . Engineered probiotics for local tumor delivery of checkpoint blockade nanobodies . Sci Transl Med . 2020 ; 12 ( 530 ): eaax0876 .

Ashenafi S , Brighenti S . Reinventing the human tuberculosis (TB) granuloma: learning from the cancer field . Front Immunol . 2022 ; 13 : 1059725 .

Forbes NS . Engineering the perfect (bacterial) cancer therapy . Nat Rev Cancer . 2010 ; 10 ( 11 ): 785 - 94 .

Yan W , Yu W , Shen L , Xiao L , Qi J , Hu T . A SARS-CoV-2 nanoparticle vaccine based on chemical conjugation of loxoribine and SpyCatcher/SpyTag . Int J Biol Macromol . 2023 ; 253 ( Pt 5 ): 127159 .

Yu D , Lu Z , Nie F , Chong Y . Integrins regulation of wound healing processes: insights for chronic skin wound therapeutics . Front Cell Infect Microbiol . 2024 ; 14 : 1324441 .

Bereta M , Hayhurst A , Gajda M , Chorobik P , Targosz M , Marcinkiewicz J , et al . Improving tumor targeting and therapeutic potential of Salmonella VNP20009 by displaying cell surface CEA-specific antibodies . Vaccine . 2007 ; 25 ( 21 ): 4183 - 92 .

Xiong X , Zheng LW , Ding Y , Chen YF , Cai YW , Wang LP , et al . Breast cancer: pathogenesis and treatments . Signal Transduct Target Ther . 2025 ; 10 ( 1 ): 49 .

Zhang Y , Huang R , Jiang Y , Shen W , Pei H , Wang G , et al . The role of bacteria and its derived biomaterials in cancer radiotherapy . Acta Pharm Sin B . 2023 ; 13 ( 10 ): 4149 - 71 .

Vincent RL , Gurbatri CR , Li F , Vardoshvili A , Coker C , Im J , et al . Probiotic-guided CAR-T cells for solid tumor targeting . Science . 2023 ; 382 ( 6667 ): 211 - 8 .

浏览量

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

工具集

关联资源

暂无数据