Strategies to package recombinant Adeno-Associated Virus expressing the N-terminal gasdermin domain for tumor treatment

• Published in: Nature communications 2021-12, Vol.12 (1), p.7155-7155, Article 7155
• Main Authors: Lu, Yuan, He, Wenbo, Huang, Xin, He, Yu, Gou, Xiaojuan, Liu, Xiaoke, Hu, Zhe, Xu, Weize, Rahman, Khaista, Li, Shan, Hu, Sheng, Luo, Jie, Cao, Gang
• Format: Article
• Language: English
• Subjects: 13/1; 13/109; 13/31; 13/44; 13/51; 14; 14/19; 14/35; 14/5; 38; 38/39; 38/47; 38/77; 38/88; 59; 631/67/1059/2325; 631/80/82; 64; 64/60; 64/86; 692/4028/67/1059/2325; Animals; Blood-brain barrier; Brain tumors; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Breast Neoplasms - physiopathology; Breast Neoplasms - therapy; Cancer; Cancer immunotherapy; Cell death; Cell Line, Tumor; Dependovirus - genetics; Dependovirus - physiology; Domains; Female; Glioblastoma; Glioblastoma - genetics; Glioblastoma - immunology; Glioblastoma - physiopathology; Glioblastoma - therapy; Humanities and Social Sciences; Humans; Immune response; Immune system; Immunotherapy; Inflammation; Insect cells; Insects; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mice, Nude; multidisciplinary; Neoplasm Proteins - genetics; Neoplasm Proteins - immunology; Oncolysis; Oncolytic Virotherapy; Oncolytic Viruses - genetics; Oncolytic Viruses - physiology; PD-L1 protein; Pore formation; Pyroptosis; Rats; Rats, Wistar; Science; Science (multidisciplinary); Sf9 Cells; Tumor cells; Tumor-infiltrating lymphocytes; Tumors; Viral Genome Packaging; Viruses
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-27407-0

Pyroptosis induced by the N-terminal gasdermin domain (GSDM NT ) holds great potential for anti-tumor therapy. However, due to the extreme cytoxicity of GSDM NT , it is challenging to efficiently produce and deliver GSDM NT into tumor cells. Here, we report the development of two strategies to package recombinant adeno-associated virus (rAAV) expressing GSDM NT : 1) drive the expression of GSDM NT by a mammal specific promoter and package the virus in Sf9 insect cells to avoid its expression; 2) co-infect rAAV-Cre to revert and express the double-floxed inverted GSDM NT . We demonstrate that these rAAVs can induce pyroptosis and prolong survival in preclinical cancer models. The oncolytic-viruses induce pyroptosis and evoke a robust immune-response. In a glioblastoma model, rAAVs temporarily open the blood-brain barrier and recruit tumor infiltrating lymphocytes into the brain. The oncolytic effect is further improved in combination with anti-PD-L1. Together, our strategies efficiently produce and deliver GSDM NT into tumor cells and successfully induce pyroptosis, which can be exploited for anti-tumor therapy. Pyroptosis, a gasdermin-mediated inflammatory cell death, could be harnessed therapeutically to improve response to cancer immunotherapy. Here the authors report the development of recombinant adeno-associated viruses to deliver the pore-forming N-terminal domain of gasdermin into cancer cells, promoting pyroptosis and anti-tumor immune responses in preclinical cancer models.