title: Proteomic screening of anaerobically regulated promoters from Salmonella and its antitumor applications.
abstract: Solid tumors often contain hypoxic and necrotic areas that can be targeted by attenuated Salmonella typhimurium VNP20009 (VNP). We sought to develop a hypoxia- inducible promoter system based on the tumor-specific delivered strain VNP to confine expression of therapeutic gene specifically or selectively within the tumor microenvironment. A hypoxia-inducible promoter - adhE promoter was screened from the hypoxia-regulated endogenous proteins of Salmonella through two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time-of-flight/time-of-flight MS-based proteomics approaches. The efficiency and specificity of the selected adhE promoter were validated first in both bacteria and animal tumor models. The adhE promoter could specifically drive GFP gene expression under hypoxia, but not under normoxia. Furthermore, luciferase reporter expression controlled by the system was also confined to the tumors. Finally, we investigated the anticancer efficacy of VNP delivering human endostatin controlled by our adhE promoter system in both murine melanoma and Lewis lung carcinoma models. Our results demonstrated that by the dual effects of tumoricidal and anti-angiogenic activities, the recombinant Salmonella strain could generate enhanced antitumor effects compared with those of unarmed VNP treatment or untreated control. The recombinant VNP could retard tumor growth significantly and extend survival of tumor-bearing mice by inducing more apoptosis and more severe necrosis as well as inhibiting blood vessel density within tumors. Therefore, VNP carrying the endostatin gene under our tumor-targeted expression system holds promise for the treatment of solid tumors.
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology and School of Stomatology, Affiliated Stomatological Hospital, Nanjing University, Nanjing, P R China.
authors.forename: Jianxiang
authors.initials: J
authors.lastname: Chen
authors.forename: Dongping
authors.initials: D
authors.lastname: Wei
authors.forename: Hongqin
authors.initials: H
authors.lastname: Zhuang
authors.forename: Yiting
authors.initials: Y
authors.lastname: Qiao
authors.forename: Bo
authors.initials: B
authors.lastname: Tang
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Wei
authors.forename: Shentong
authors.initials: S
authors.lastname: Fang
authors.forename: Guo
authors.initials: G
authors.lastname: Chen
authors.forename: Pan
authors.initials: P
authors.lastname: Du
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Huang
authors.forename: Wenhui
authors.initials: W
authors.lastname: Jiang
authors.forename: Qingang
authors.initials: Q
authors.lastname: Hu
authors.forename: Zi-Chun
authors.initials: ZC
authors.lastname: Hua
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000693:Anaerobiosis; D020533:Angiogenesis Inhibitors / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001691:Biological Therapy / Q000379:methods*; D018827:Carcinoma, Lewis Lung / Q000473:pathology / Q000628:therapy; D016923:Cell Death; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply; D043169:Endostatins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D053208:Kaplan-Meier Estimate; D008546:Melanoma, Experimental / Q000473:pathology / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation; D030781:Organisms, Genetically Modified; D011401:Promoter Regions, Genetic; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012486:Salmonella typhimurium / Q000235:genetics / Q000378:metabolism*; D047368:Tumor Burden; D015854:Up-Regulation
chemical_list: D020533:Angiogenesis Inhibitors; D001426:Bacterial Proteins; D043169:Endostatins; D020543:Proteome; D011993:Recombinant Fusion Proteins; D000426:Alcohol Dehydrogenase