Construction of recombinant oncolytic vaccinia virus armed with anti-FAP and anti-CD3 bi-specific antibody and its anti-tumor activity
SHI Jin-sheng1, LI Feng1, WU Si-hui2, YU Li-chao1, LIU Li-qiong3,YUFeng4,5
(1. Department of Cardiothoracic Surgery,Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001; 2. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 3. Department of Hematology,Shenzhen Nanshan Hospital, Shenzhen Guangdong 5180052; 4. Institute of Life Sciences, Jiangsu University,Zhenjiang Jiangsu 212013; 5. the State Key Laboratory of Oncogenes and Related Genes,Shanghai 200032, China)
Abstract：Objective: To construct double deleting recombinant oncolytic vaccinia virus armed with bi-specific antibody of antifibroblast activation protein(FAP) and anti-mouse CD3(BiTE.FAP), and to evaluate the anti-tumor efficacy in vitro. Methods: The BiTE-FAP gene regulated by the F17R promoter(PF17R) and the DsRed gene regulated by the SE/L promoter(PSEL) were homologously recombinated into the TK locus of the vSC20 parental virus to construct the recombinant double deleted vaccinia virus-bispecific T cell engager-FAP(rVVDD- BiTE.FAP).Plaque counting was used to assess the replication ability of recombinant oncolytic virus. MTS and LDH assay were used to detect the inhibitory effect on mouse lung tumor cells.ELISA assay was performed to analyse the IL-2 and IFN-γ expression by mouse spleen T cells. Results: rVVDD-BiTE.FAP was obtained by homologous recombination and soft agar purification.MTS and crystal violet staining results showed that rVVDD-BiTE.FAP has the same replication capacity compared with the parental vSC20 virus and mouse T cells could enhance the anti-tumor effect of rVVDD-BiTE.FAP. Conclusion: rVVDD-BiTE.FAP has high anti-tumor efficacy.
施晋升1， 李峰1， 吴思慧2， 俞力超1， 刘黎琼3， 于峰4,5. 携带抗FAP×抗CD3双特异性抗体重组溶瘤痘病毒的构建及其抗癌活性观察[J]. 江苏大学学报:医学版, 2019, 29(04): 327-332.
SHI Jin-sheng1, LI Feng1, WU Si-hui2, YULi-chao1, LIU Li-qiong3,YU Feng4,5.
Construction of recombinant oncolytic vaccinia virus armed with anti-FAP and anti-CD3 bi-specific antibody and its anti-tumor activity. Journal of Jiangsu University(Medicine Edition), 2019, 29(04): 327-332.
［1］Wu JS,Sheng SR,Liang XH,et al.The role of tumor microenvironment in collective tumor cell invasion\[J\].Future Oncol,2017,13(11):991-1002.［2］Fuming ZI,Jingsong HE,Donghua HE,et al.Fibroblast activation protein α in tumor microenvironment:Recent progression and implications(Review)\[J\].Mol Med Rep,2015, 11(5):3203-3211.［3］Gottschalk S,Yu F,Ji M,et al.A vaccine that cotargets tumor cells and cancer associated fibroblasts results in enhanced antitumor activity by inducing antigen spreading\[J\].PLoS One,2013,8(12):e82658.［4］Wong HH,Lemoine NR,Wang Y.Oncolytic viruses for cancer therapy:Overcoming the obstacles\[J\].Viruses,2010,2(1):78-106.［5］Hughes J,Wang P,Alusi G,et al.Lister strain vaccinia virus with thymidine kinase gene deletion is a tractable platform for development of a new generation of oncolytic virus\[J\]. Gene Ther,2015,22(6):476-484.［6］Yamaguchi T,Uchida E.Oncolytic virus: regulatory aspects from quality control to clinical studies\[J\].Curr Cancer Drug Targets,2018，18(2):202-208.［7］Marx J.Cancer biology.All in the stroma: cancer′s cosa nostra\[J\].Science, 2008,320(5872): 38-41.［8］Chen M, Xiang R, Wen Y, et al. A wholecell tumor vaccine modified to express fibroblast activation protein induces antitumor immunity against both tumor cells and cancerassociated fibroblasts\[J\].Sci Rep,2015,5:14421.［9］Akinboye ES, Brennen WN, Rosen DM, et al. Iterative design of emetinebased prodrug targeting fibroblast activation protein(FAP) and dipeptidyl peptidase IV DPPIV using a tandem enzymatic activation strategy\[J\].Prostate, 2016,76(8):703-714.［10］Christiansen VJ,Jackson KW,Lee KN,et al.Targeting inhibition of fibroblast activation proteinα and prolyl oligopeptidase activities on cells common to metastatic tumor microenvironments\[J\]. Neoplasia,2013,15(4): 348-358.［11］Li M, Li M,Yin T,et al.Targeting of cancer associated fibroblasts enhances the efficacy of cancer chemotherapy by regulating the tumor microenvironment\[J\].Mol Med Rep, 2016,13(3): 2476-2484.［12］Jiang GM,Xu W,Du J,et al.The application of the fibroblast activation protein αtargeted immunotherapy strategy\[J\].Oncotarget,2016,7(22):33472-33482.［13］Ostermann E,Garinchesa P,Heider KH,et al. Effective immunoconjugate therapy in cancer models targeting a serine protease of tumor fibroblasts\[J\].Clin Cancer Res,2008, 14(14):4584-4592.［14］Cunningham CC.Talabostat\[J\].Expert Opin Investig Drugs,2007,16(9):1459-1465.［15］Tran E, Chinnasamy D,Yu Z, et al.Immune targeting of fibroblast activation protein triggers recognition of multipotent bone marrow stromal cells and cachexia\[J\].J Exp Med, 2013,210(6):1125-1135.
2019, Vol. 29 
