Rapid screening of engineered microbial therapies in a 3D multicellular model
| Autor: | Taylor E. Hinchliffe, Samuel Castro, Oscar Velazquez, William H. Mather, Zakary S. Singer, Tal Danino, Tetsuhiro Harimoto, Joanna Zhang |
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| Rok vydání: | 2019 |
| Předmět: |
Salmonella typhimurium
High-throughput screening Computational biology Biology 03 medical and health sciences Synthetic biology Mice 0302 clinical medicine Synthetic gene Spheroids Cellular Diagnosis Escherichia coli Animals Gene Regulatory Networks Proteus mirabilis Diagnostic Techniques and Procedures 030304 developmental biology 0303 health sciences Multidisciplinary Disease progression Translation (biology) Biological Sciences Biocontainment Listeria monocytogenes In vitro Coculture Techniques High-Throughput Screening Assays Multicellular organism Disease Models Animal 030220 oncology & carcinogenesis Tumor reduction Multicellular spheroid Syngeneic mouse Synthetic Biology Drug Screening Assays Antitumor Genetic Engineering |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 116(18) |
| ISSN: | 1091-6490 |
| Popis: | Synthetic biology is transforming therapeutic paradigms by engineering living cells and microbes to intelligently sense and respond to diseases including inflammation1,2, infections3-5, metabolic disorders6,7, and cancer8,9. However, the ability to rapidly engineer new therapies far outpaces the throughput of animal-based testing regimes, creating a major bottleneck for clinical translation10,11. In vitro approaches to address this challenge have been limited in scalability and broad-applicability. Here, we present a bacteria-in-spheroid co-culture (BSCC) platform that simultaneously tests host species, therapeutic payloads and synthetic gene circuits of engineered bacteria within multicellular spheroids over a timescale of weeks. Long-term monitoring of bacterial dynamics and disease progression enables quantitative comparison of critical therapeutic parameters such as efficacy and biocontainment. Specifically, we screen S. typhimurium strains expressing and delivering a library of antitumor therapeutic molecules via several synthetic gene circuits. We identify novel candidates exhibiting significant tumor reduction and demonstrate high similarity in their efficacies using a syngeneic mouse model. Lastly, we show that our platform can be expanded to dynamically profile diverse microbial species including L. monocytogenes, P. mirabilis, and E. coli in various host cell types. This high-throughput framework may serve to accelerate synthetic biology for clinical applications and understanding the host-microbe interactions in disease sites. |
| Databáze: | OpenAIRE |
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