Yutaro Kubota, Ming Zhao, Qinghong Han, Yusuke Aoki, Noriyuki Masaki, Koya Obara, Sei Morinaga, Kohei Mizuta, Motokazu Sato, Michael Bouvet, Koichi Kubota, Takuya Tsunoda, Robert M Hoffman
{"title":"表达肿瘤靶向鼠伤寒沙门菌A1-R的工程蛋氨酸酶通过消耗肿瘤蛋氨酸抑制同基因肿瘤小鼠模型。","authors":"Yutaro Kubota, Ming Zhao, Qinghong Han, Yusuke Aoki, Noriyuki Masaki, Koya Obara, Sei Morinaga, Kohei Mizuta, Motokazu Sato, Michael Bouvet, Koichi Kubota, Takuya Tsunoda, Robert M Hoffman","doi":"10.21873/cgp.20499","DOIUrl":null,"url":null,"abstract":"<p><strong>Background/aim: </strong>We previously developed <i>Salmonella typhimurium</i> A1-R, which selectively targets and kills tumors. In the present study, we established recombinant methioninase (rMETase)-producing <i>Salmonella typhimurium</i> A1-R (A1-R-rMETase), by transfer of the <i>Pseudomonas putida methioninase</i> gene, to target methionine addiction of syngeneic-cancer mouse models.</p><p><strong>Materials and methods: </strong>A plasmid containing the <i>Pseudomonas putida methioninase</i> gene was extracted from METase-producing recombinant <i>E. coli</i> and inserted into <i>Salmonella typhimurium</i> A1-R using electroporation. Lewis Lung Carcinoma (LLC) cells (10<sup>6</sup>) were injected subcutaneously in male C57BL/6 mice aged 4-6 weeks. We determined that 10<sup>8</sup> <i>Salmonella typhimurium</i> A1-R-rMETase administered iv was a safe dosage in C57BL/6 mice and was used for efficacy studies on LLC tumors in C57BL/6 mice. Tumor size was measured with calipers three times per week for 3 weeks. On day 22, tumor methionine levels were measured using HPLC in the control mice injected with phosphate-buffered saline (PBS) and the mice injected with <i>Salmonella typhimurium</i> A1-R-rMETase.</p><p><strong>Results: </strong>The mean LLC tumor size of each group on day 22 was as follows: PBS control: 741.5 mm<sup>3</sup>; mice injected with <i>Salmonella typhimurium</i> A1-R: 566.3 mm<sup>3</sup> (<i>p</i>=0.370); and mice injected with <i>Salmonella typhimurium</i> A1-R-rMETase: 198.8 mm<sup>3</sup> (<i>p</i>=0.0003 <i>vs</i> control and <i>p</i>=0.0117 <i>vs. Salmonella</i> A1-R). The mice injected with <i>Salmonella typhimurium</i> A1-R-rMETase showed a significantly lower mean tumor methionine level than mice injected with PBS (5.9 nM/mg protein <i>vs</i> 11.1 nM/mg protein, <i>p</i>=0.0095). <i>Salmonella typhimurium</i> A1-R-rMETase grew continuously in the tumors but not in the liver or spleen.</p><p><strong>Conclusion: </strong>Tumor-targeting <i>Salmonella typhimurium</i> A1-R engineered to express the <i>Pseudomonas putida methioninase</i> gene, inhibited LLC tumor growth in a syngeneic mouse model and reduced the methionine level in the tumor. <i>Salmonella typhimurium</i> A1-R-rMETase combines the tumor targeting and killing capability of <i>Salmonella typhimurium</i> A1-R plus rMETase which targets the methionine addiction of cancer.</p>","PeriodicalId":9516,"journal":{"name":"Cancer Genomics & Proteomics","volume":"22 2","pages":"247-257"},"PeriodicalIF":2.6000,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880925/pdf/","citationCount":"0","resultStr":"{\"title\":\"Engineered Methioninase-expressing Tumor-targeting <i>Salmonella typhimurium</i> A1-R Inhibits Syngeneic-Cancer Mouse Models by Depleting Tumor Methionine.\",\"authors\":\"Yutaro Kubota, Ming Zhao, Qinghong Han, Yusuke Aoki, Noriyuki Masaki, Koya Obara, Sei Morinaga, Kohei Mizuta, Motokazu Sato, Michael Bouvet, Koichi Kubota, Takuya Tsunoda, Robert M Hoffman\",\"doi\":\"10.21873/cgp.20499\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background/aim: </strong>We previously developed <i>Salmonella typhimurium</i> A1-R, which selectively targets and kills tumors. In the present study, we established recombinant methioninase (rMETase)-producing <i>Salmonella typhimurium</i> A1-R (A1-R-rMETase), by transfer of the <i>Pseudomonas putida methioninase</i> gene, to target methionine addiction of syngeneic-cancer mouse models.</p><p><strong>Materials and methods: </strong>A plasmid containing the <i>Pseudomonas putida methioninase</i> gene was extracted from METase-producing recombinant <i>E. coli</i> and inserted into <i>Salmonella typhimurium</i> A1-R using electroporation. Lewis Lung Carcinoma (LLC) cells (10<sup>6</sup>) were injected subcutaneously in male C57BL/6 mice aged 4-6 weeks. We determined that 10<sup>8</sup> <i>Salmonella typhimurium</i> A1-R-rMETase administered iv was a safe dosage in C57BL/6 mice and was used for efficacy studies on LLC tumors in C57BL/6 mice. Tumor size was measured with calipers three times per week for 3 weeks. On day 22, tumor methionine levels were measured using HPLC in the control mice injected with phosphate-buffered saline (PBS) and the mice injected with <i>Salmonella typhimurium</i> A1-R-rMETase.</p><p><strong>Results: </strong>The mean LLC tumor size of each group on day 22 was as follows: PBS control: 741.5 mm<sup>3</sup>; mice injected with <i>Salmonella typhimurium</i> A1-R: 566.3 mm<sup>3</sup> (<i>p</i>=0.370); and mice injected with <i>Salmonella typhimurium</i> A1-R-rMETase: 198.8 mm<sup>3</sup> (<i>p</i>=0.0003 <i>vs</i> control and <i>p</i>=0.0117 <i>vs. Salmonella</i> A1-R). The mice injected with <i>Salmonella typhimurium</i> A1-R-rMETase showed a significantly lower mean tumor methionine level than mice injected with PBS (5.9 nM/mg protein <i>vs</i> 11.1 nM/mg protein, <i>p</i>=0.0095). <i>Salmonella typhimurium</i> A1-R-rMETase grew continuously in the tumors but not in the liver or spleen.</p><p><strong>Conclusion: </strong>Tumor-targeting <i>Salmonella typhimurium</i> A1-R engineered to express the <i>Pseudomonas putida methioninase</i> gene, inhibited LLC tumor growth in a syngeneic mouse model and reduced the methionine level in the tumor. <i>Salmonella typhimurium</i> A1-R-rMETase combines the tumor targeting and killing capability of <i>Salmonella typhimurium</i> A1-R plus rMETase which targets the methionine addiction of cancer.</p>\",\"PeriodicalId\":9516,\"journal\":{\"name\":\"Cancer Genomics & Proteomics\",\"volume\":\"22 2\",\"pages\":\"247-257\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2025-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11880925/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cancer Genomics & Proteomics\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.21873/cgp.20499\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"GENETICS & HEREDITY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cancer Genomics & Proteomics","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.21873/cgp.20499","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
Background/aim: We previously developed Salmonella typhimurium A1-R, which selectively targets and kills tumors. In the present study, we established recombinant methioninase (rMETase)-producing Salmonella typhimurium A1-R (A1-R-rMETase), by transfer of the Pseudomonas putida methioninase gene, to target methionine addiction of syngeneic-cancer mouse models.
Materials and methods: A plasmid containing the Pseudomonas putida methioninase gene was extracted from METase-producing recombinant E. coli and inserted into Salmonella typhimurium A1-R using electroporation. Lewis Lung Carcinoma (LLC) cells (106) were injected subcutaneously in male C57BL/6 mice aged 4-6 weeks. We determined that 108Salmonella typhimurium A1-R-rMETase administered iv was a safe dosage in C57BL/6 mice and was used for efficacy studies on LLC tumors in C57BL/6 mice. Tumor size was measured with calipers three times per week for 3 weeks. On day 22, tumor methionine levels were measured using HPLC in the control mice injected with phosphate-buffered saline (PBS) and the mice injected with Salmonella typhimurium A1-R-rMETase.
Results: The mean LLC tumor size of each group on day 22 was as follows: PBS control: 741.5 mm3; mice injected with Salmonella typhimurium A1-R: 566.3 mm3 (p=0.370); and mice injected with Salmonella typhimurium A1-R-rMETase: 198.8 mm3 (p=0.0003 vs control and p=0.0117 vs. Salmonella A1-R). The mice injected with Salmonella typhimurium A1-R-rMETase showed a significantly lower mean tumor methionine level than mice injected with PBS (5.9 nM/mg protein vs 11.1 nM/mg protein, p=0.0095). Salmonella typhimurium A1-R-rMETase grew continuously in the tumors but not in the liver or spleen.
Conclusion: Tumor-targeting Salmonella typhimurium A1-R engineered to express the Pseudomonas putida methioninase gene, inhibited LLC tumor growth in a syngeneic mouse model and reduced the methionine level in the tumor. Salmonella typhimurium A1-R-rMETase combines the tumor targeting and killing capability of Salmonella typhimurium A1-R plus rMETase which targets the methionine addiction of cancer.
期刊介绍：
Cancer Genomics & Proteomics (CGP) is an international peer-reviewed journal designed to publish rapidly high quality articles and reviews on the application of genomic and proteomic technology to basic, experimental and clinical cancer research. In this site you may find information concerning the editorial board, editorial policy, issue contents, subscriptions, submission of manuscripts and advertising. The first issue of CGP circulated in January 2004.
Cancer Genomics & Proteomics is a journal of the International Institute of Anticancer Research. From January 2013 CGP is converted to an online-only open access journal.
Cancer Genomics & Proteomics supports (a) the aims and the research projects of the INTERNATIONAL INSTITUTE OF ANTICANCER RESEARCH and (b) the organization of the INTERNATIONAL CONFERENCES OF ANTICANCER RESEARCH.
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