<i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method
We demonstrate that <i>Blautia coccoides</i> JCM1395<sup>T</sup> has the potential to be used for tumor-targeted live bacterial therapeutics. Prior to studying its in vivo biodistribution, a sample preparation method for reliable quantitative analysis of bacteria in biologica...
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| Format: | Article |
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MDPI AG
2023-03-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/15/3/989 |
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| author | Shoko Nomura Erike W. Sukowati Yuko Shigeno Maiko Takahashi Akari Kato Yoshimi Benno Fumiyoshi Yamashita Hidefumi Mukai |
| author_facet | Shoko Nomura Erike W. Sukowati Yuko Shigeno Maiko Takahashi Akari Kato Yoshimi Benno Fumiyoshi Yamashita Hidefumi Mukai |
| author_sort | Shoko Nomura |
| collection | DOAJ |
| description | We demonstrate that <i>Blautia coccoides</i> JCM1395<sup>T</sup> has the potential to be used for tumor-targeted live bacterial therapeutics. Prior to studying its in vivo biodistribution, a sample preparation method for reliable quantitative analysis of bacteria in biological tissues was required. Gram-positive bacteria have a thick outer layer of peptidoglycans, which hindered the extraction of 16S rRNA genes for colony PCR. We developed the following method to solve the issue; the method we developed is as follows. The homogenates of the isolated tissue were seeded on agar medium, and bacteria were isolated as colonies. Each colony was heat-treated, crushed with glass beads, and further treated with restriction enzymes to cleave DNAs for colony PCR. With this method, <i>Blautia coccoides</i> JCM1395<sup>T</sup> and <i>Bacteroides vulgatus</i> JCM5826<sup>T</sup> were individually detected from tumors in mice intravenously receiving their mixture. Since this method is very simple and reproducible, and does not involve any genetic modification, it can be applied to exploring a wide range of bacterial species. We especially demonstrate that <i>Blautia coccoides</i> JCM1395<sup>T</sup> efficiently proliferate in tumors when intravenously injected into tumor-bearing mice. Furthermore, these bacteria showed minimal innate immunological responses, i.e., elevated serum tumor necrosis factor α and interleukin-6, similar to <i>Bifidobacterium</i> sp., which was previously studied as a therapeutic agent with a small immunostimulating effect. |
| first_indexed | 2024-03-11T06:01:05Z |
| format | Article |
| id | doaj.art-4a403dae5d8d4d9f9594512680e53f54 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-11T06:01:05Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-4a403dae5d8d4d9f9594512680e53f542023-11-17T13:17:14ZengMDPI AGPharmaceutics1999-49232023-03-0115398910.3390/pharmaceutics15030989<i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR MethodShoko Nomura0Erike W. Sukowati1Yuko Shigeno2Maiko Takahashi3Akari Kato4Yoshimi Benno5Fumiyoshi Yamashita6Hidefumi Mukai7Laboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe 650-0047, JapanLaboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe 650-0047, JapanBenno Laboratory, RIKEN Baton Zone Program, RIKEN Cluster for Science Technology and Innovation Hab, Wako 351-0198, JapanLaboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe 650-0047, JapanLaboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe 650-0047, JapanBenno Laboratory, RIKEN Baton Zone Program, RIKEN Cluster for Science Technology and Innovation Hab, Wako 351-0198, JapanDepartment of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, JapanLaboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe 650-0047, JapanWe demonstrate that <i>Blautia coccoides</i> JCM1395<sup>T</sup> has the potential to be used for tumor-targeted live bacterial therapeutics. Prior to studying its in vivo biodistribution, a sample preparation method for reliable quantitative analysis of bacteria in biological tissues was required. Gram-positive bacteria have a thick outer layer of peptidoglycans, which hindered the extraction of 16S rRNA genes for colony PCR. We developed the following method to solve the issue; the method we developed is as follows. The homogenates of the isolated tissue were seeded on agar medium, and bacteria were isolated as colonies. Each colony was heat-treated, crushed with glass beads, and further treated with restriction enzymes to cleave DNAs for colony PCR. With this method, <i>Blautia coccoides</i> JCM1395<sup>T</sup> and <i>Bacteroides vulgatus</i> JCM5826<sup>T</sup> were individually detected from tumors in mice intravenously receiving their mixture. Since this method is very simple and reproducible, and does not involve any genetic modification, it can be applied to exploring a wide range of bacterial species. We especially demonstrate that <i>Blautia coccoides</i> JCM1395<sup>T</sup> efficiently proliferate in tumors when intravenously injected into tumor-bearing mice. Furthermore, these bacteria showed minimal innate immunological responses, i.e., elevated serum tumor necrosis factor α and interleukin-6, similar to <i>Bifidobacterium</i> sp., which was previously studied as a therapeutic agent with a small immunostimulating effect.https://www.mdpi.com/1999-4923/15/3/989bacteriabacterial cancer therapycolony PCR16S rRNA genes<i>Blautia coccoides</i> |
| spellingShingle | Shoko Nomura Erike W. Sukowati Yuko Shigeno Maiko Takahashi Akari Kato Yoshimi Benno Fumiyoshi Yamashita Hidefumi Mukai <i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method Pharmaceutics bacteria bacterial cancer therapy colony PCR 16S rRNA genes <i>Blautia coccoides</i> |
| title | <i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method |
| title_full | <i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method |
| title_fullStr | <i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method |
| title_full_unstemmed | <i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method |
| title_short | <i>Blautia coccoides</i> JCM1395<sup>T</sup> Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method |
| title_sort | i blautia coccoides i jcm1395 sup t sup achieved intratumoral growth with minimal inflammation evidence for live bacterial therapeutic potential by an optimized sample preparation and colony pcr method |
| topic | bacteria bacterial cancer therapy colony PCR 16S rRNA genes <i>Blautia coccoides</i> |
| url | https://www.mdpi.com/1999-4923/15/3/989 |
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