Therapeutic Effect of Recombinant Attenuated Listeria monocytogenes Expressing E7 Protein in a Mouse (Mus musculus) Model of Cervical Cancer
PAN Xiao-Xin, CUI Ming-Zhu, LUO Ya-Ru, SUN Jing, CHENG Chang-Yong, WANG Jing*, SONG Hou-Hui*
Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Hangzhou 311300, China
AbstractListeria monocytogenes is a high-quality vaccine candidate live vector that can migrate between cells and stimulate the body to promote the proliferation of antigen-specific CD4+ and CD8+ T cells to produce adaptive cellular immunity. This research group found that attenuated Listeria monocytogenes (LAM) had significantly lower hemolytic and proliferative ability than wild-type Listeria. At the same time, LAM could normally secrete Listeriolysin O (LLO) protein. In order to study the safety of LAM and its delivery and immune effect on the cervical cancer specific antigen E7 protein, this study tested the virulence of LAM in a mouse (Mus musculus) model, and to evaluate the role of attenuated Listeria monocytogenes-E7 (LAM-E7) in the immunotherapy and prevention of cervical cancer in mice. The results showed that the virulence of LAM to mice was 1/8709 of that of wild-type Listeria, and LAM could be eliminated from the organs of mice within a week. In this study, E7 gene was integrated into LAM genome by homologous recombination method. PCR identification results showed that LAM-E7 strain was successfully constructed, and Western blot results showed that E7 protein could be fused with LLO and expressed and released in the recombinant strain. The analysis of the growth ability and hemolytic activity of LAM-E7 in vitro showed that the fusion had no significant effect on the growth ability and hemolytic ability of LAM, indicating that the fusion of E7 protein would not affect the safety of LAM. In the cervical cancer treatment trial, the average tumor volume of the LAM-E7 group (161.1202 mm3) was notably smaller than that of the LAM group (349.4625 mm3) and the PBS group (512.4149 mm3)(P<0.01), indicating a significant inhibitory effected on tumor growth. The immunoprevention test showed that the average tumor volume of the LAM-E7 group (3.86 mm3) was significantly smaller than that of the LAM group (172.67 mm3) and the PBS group (57.38 mm3)(P<0.001), demonstrating the preventive effect of LAM-E7 on cervical cancer. LAM-E7 has demonstrated potent therapeutic and preventive properties on cervical cancer tumors in mice, which offers a new perspective for using attenuated Listeria as a vaccine vector in the treatment of cervical cancer. This study also provides valuable data that supports the advancement of therapeutic vaccines for cervical cancer in the future.
PAN Xiao-Xin,CUI Ming-Zhu,LUO Ya-Ru, et al. Therapeutic Effect of Recombinant Attenuated Listeria monocytogenes Expressing E7 Protein in a Mouse (Mus musculus) Model of Cervical Cancer[J]. 农业生物技术学报, 2024, 32(5): 1206-1215.
[1] 贾艳艳, 刘莎莎, 汪洋, 等. 2016. 减毒单增李斯特菌载体在肿瘤疫苗中的应用[J]. 中国免疫学杂志, 32: 1866-1869. (Jia Y Y, Liu S S, Wang Y, et al.2016. Application of attenuated Listeria monocytogenes vector in tumor vaccines[J] Chinese Journal of Immunology, 32: 1866-1869.) [2] 见永康, 吴泽俊, 何娟, 等. 2022. 人乳头瘤病毒E6/E7 mRNA在宫颈病变中的研究进展[J]. 癌症进展, 1(2): 113-115. (Jian Y K, Wu Z J, He J, et al.2022. Research progress of Human papillomavirus E6/E7 mRNA in cervical lesions[J]. Progression of Cancer, 1(2): 113-115.) [3] Chakraborty E, Sarkar D.2022. Emerging therapies for hepatocellular carcinoma (HCC)[J]. Cancers (Basel), 14(11):2798-2821. [4] Chávez-Arroyo A, Portnoy D A.2020. Why is Listeria monocytogenes such a potent inducer of CD8+ T-cells?[J]. Cellular Microbiology, 22(4): 13175-13185. [5] Farmer E, Cheng M A, Hung C F, et al.2021. Vaccination strategies for the control and treatment of HPV infection and HPV-associated cancer[J]. Recent Results In Cancer Research, 217: 157-195. [6] Flickinger J C Jr, Rodeck U, Snook A E.2018. Listeria monocytogenes as a vector for cancer immunotherapy: Current understanding and progress[J]. Vaccines (Basel), 6(3):48-66. [7] Gahan C G, Hill C.2014. Listeria monocytogenes: Survival and adaptation in the gastrointestinal tract[J]. Frontiers in Cellular and Infection Microbiology, 4: 9-16. [8] Garcia J, Hurwitz HI, Sandler AB, et al.2020. Bevacizumab (Avastin®) in cancer treatment: A review of 15 years of clinical experience and future outlook[J]. Cancer Treatment Reviews, 86: 102017. [9] Gravekamp C, Paterson Y.2010. Harnessing Listeria monocytogenes to target tumors[J]. Cancer Biology & Therapy,. 9(4): 257-265. [10] Henriques A R, Cristino J M, Fraqueza M J.2017. Genetic characterization of Listeria monocytogenes isolates from industrial and retail ready-to-eat meat-based foods and their relationship with clinical strains from human Listeriosis in portugal[J]. Journal of Food Protection, 80(4): 551-560. [11] Hoppe-Seyler K, Bossler F, Braun J A, et al.2018. The HPV E6/E7 oncogenes: Key factors for viral carcinogenesis and therapeutic targets[J]. Trends in Microbiology, 26(2): 158-168. [12] Kaptchouang Tchatchouang C D, Fri J, Santi M D, et al.2020. Listeriosis outbreak in South Africa: A comparative analysis with previously reported cases worldwide[J]. Microorganisms, 8(1): 135-151. [13] Opperman C J, Bamford C.2018. Co-infection with streptococcus pneumoniae and Listeria monocytogenes in an immunocompromised patient[J]. South Afrecan Medical Journal, 108(5): 386-388. [14] Parkin D M, Bray F, Ferlay J, et al.2005. Global cancer statistics, 2002[J]. CA-A Cancer Journal for Clinicians, 55(2): 74-108. [15] Olusola P, Banerjee H N, Philley J V, et al.2019. Human papilloma virus-associated cervical cancer and health disparities[J]. Cells, 8(6): 622-634. [16] Sciaranghella G, Lakhashe SK, Ayash-Rashkovsky M, et al.2011. A live attenuated Listeria monocytogenes vaccine vector expressing SIV Gag is safe and immunogenic in macaques and can be administered repeatedly[J]. Vaccine, 29(3): 476-486. [17] Shen H, Slifka M K, Matloubian M, et al.1995. Recombinant Listeria monocytogenes as a live vaccine vehicle for the induction of protective anti-viral cell-mediated immunity[J]. Proceedings of the National Academy of Sciences of the USA, 92(9): 3987-3991. [18] Starks H, Bruhn K W, Shen H, et al.2004. Listeria monocytogenes as a vaccine vector: Virulence attenuation or existing antivector immunity does not diminish therapeutic efficacy[J]. The Journal of Immunology, 173(1):420-427. [19] Vazquez-Boland J A, Krypotou E, Scortti M.2017. Listeria placental infection[J]. MBio, 8(3): e00949-e01017. [20] Woo S R, Fuertes M B, Corrales L, et al.2014. STING-dependent cytosolic DNA sensing mediates innate immune recognition of immunogenic tumors[J]. Immunity, 41(5): 830-842. [21] Wan X, Cheng C, Lin Z, et al.2015. The attenuated hepatocellular carcinoma-specific Listeria vaccine Lmdd-MPFG prevents tumor occurrence through immune regulation of dendritic cells[J]. Oncotarget, 6(11): 8822-8838. [22] Yang A, Farmer E, Wu T C, et al.2016. Perspectives for therapeutic HPV vaccine development[J]. Journal of Biomedical Science, 23(1): 75-94. [23] Yim E K, Park J S.2005. The role of HPV E6 and E7 oncoproteins in HPV-associated cervical carcinogenesis[J]. Cancer Research and Treatment, 37(6): 319-324. [24] Zhao X, Yang F, Mariz F, et al.2020. Combined prophylactic and therapeutic immune responses against human papillomaviruses induced by a thioredoxin-based L2-E7 nanoparticle vaccine[J]. PLOS Pathogens, 16(9): 1008827-1008849. [25] Zhou S, Gravekamp C, Bermudes D, et al.2018. Tumour-targeting bacteria engineered to fight cancer[J]. Nature Reviews Cancer, 18(12): 727-743.
