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Pharmaceutics
Volume 15
Issue 3
10.3390/pharmaceutics15030989
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Article

Blautia coccoides JCM1395T Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method

by
Shoko Nomura
1,2,
Erike W. Sukowati
1,
Yuko Shigeno
3,
Maiko Takahashi
1,
Akari Kato
1,
Yoshimi Benno
3,
Fumiyoshi Yamashita
2 and
Hidefumi Mukai
1,4,*
1
Laboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe 650-0047, Japan
2
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
3
Benno Laboratory, RIKEN Baton Zone Program, RIKEN Cluster for Science Technology and Innovation Hab, Wako 351-0198, Japan
4
Department of Pharmaceutical Informatics, Graduate School of Biomedical Science, Nagasaki University, Nagasaki 852-8588, Japan
*
Author to whom correspondence should be addressed.
Pharmaceutics 2023, 15(3), 989; https://doi.org/10.3390/pharmaceutics15030989
Submission received: 24 January 2023 / Revised: 4 March 2023 / Accepted: 17 March 2023 / Published: 19 March 2023
(This article belongs to the Special Issue Novel Cell and Bioinspired Drug Delivery Systems)
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Abstract

We demonstrate that Blautia coccoides JCM1395T 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, Blautia coccoides JCM1395T and Bacteroides vulgatus JCM5826T 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 Blautia coccoides JCM1395T 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 Bifidobacterium sp., which was previously studied as a therapeutic agent with a small immunostimulating effect.
Keywords: bacteria; bacterial cancer therapy; colony PCR; 16S rRNA genes; Blautia coccoides bacteria; bacterial cancer therapy; colony PCR; 16S rRNA genes; Blautia coccoides

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MDPI and ACS Style

Nomura, S.; Sukowati, E.W.; Shigeno, Y.; Takahashi, M.; Kato, A.; Benno, Y.; Yamashita, F.; Mukai, H. Blautia coccoides JCM1395T Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method. Pharmaceutics 2023, 15, 989. https://doi.org/10.3390/pharmaceutics15030989

AMA Style

Nomura S, Sukowati EW, Shigeno Y, Takahashi M, Kato A, Benno Y, Yamashita F, Mukai H. Blautia coccoides JCM1395T Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method. Pharmaceutics. 2023; 15(3):989. https://doi.org/10.3390/pharmaceutics15030989

Chicago/Turabian Style

Nomura, Shoko, Erike W. Sukowati, Yuko Shigeno, Maiko Takahashi, Akari Kato, Yoshimi Benno, Fumiyoshi Yamashita, and Hidefumi Mukai. 2023. "Blautia coccoides JCM1395T Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method" Pharmaceutics 15, no. 3: 989. https://doi.org/10.3390/pharmaceutics15030989

APA Style

Nomura, S., Sukowati, E. W., Shigeno, Y., Takahashi, M., Kato, A., Benno, Y., Yamashita, F., & Mukai, H. (2023). Blautia coccoides JCM1395T Achieved Intratumoral Growth with Minimal Inflammation: Evidence for Live Bacterial Therapeutic Potential by an Optimized Sample Preparation and Colony PCR Method. Pharmaceutics, 15(3), 989. https://doi.org/10.3390/pharmaceutics15030989

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