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Team:Munich/Safety

Phactory

Safety

General Laboratory Safety

The iGEM Team Munich is in full compliance with the safety and security rules of the iGEM competition.

The team was included in the laboratories of Friedrich Simmel (Technical University of Munich) and Gil Westmeyer (Helmholtz Zentrum München), which fall under the biosafety level 1. This level is the lowest biosafety safety level and was sufficient for all our experiments. Before we started our work in the lab, all team members had to pass different safety tests and receive training during an introductory wetlab safety week. Those tests included an introduction in the laboratory machinery (i.e. thermocycler, plate reader, gel chamber), but also covered the use of chemicals (i.e. chloroform, phenol, staining reagents). Moreover, we were required to participate in the mandatory university security introductions, including explanation of reanimation equipment, first aid and escape routes. At all times, a supervisor or instructor was present when work was conducted in the laboratory.

Project safety

The overuse of antibiotics is leading humanity in a dangerous direction. The WHO calculated that 900,000 resistant infections occure in hospitals annually1, killing 63,000 patients2. Every year, antimicrobial resistance results in 8 million more hospitalization days, adding more than $20 billion to US healthcare costs3. The use of antibiotics is highly associated with a reduction of microbiome diversity, which in turn is related several types diseases4. Therefore, it is essential to find other forms of therapy for bacterial infections. Over the last years, phage therapy has been re-discovered because of the promising outcomes achieved in Eastern Europe5. Now, the Western countries have started to focus on this field by conducting important research with studies like Phage4Cure. Our cell-free assembly platform increases safety and supports the field in the following ways:

Overcoming the Antibiotics Crisis with Phages

Phages have been proven to work on pathogens resistant to antibiotics6. By producing therapeutic phages, the treatment repertoire is increased. This opens up the possibility to cure patients where conventional treatment has failed7.

Time-Saving Production of Phages Can Save a Patient’s Life

We have shown that we can assemble a phage of our choice in 3 hours, whereas the native isolation of the same phage takes at least 6 hours. With regard to the treatment of a patient, this time difference can be lifesaving.

Phages are More Specific than Antibiotics:

Antibiotics target abundant characteristics of bacteria. Thus, they can be used against a broad range bacteria8, however also impacting the human microbiome9. Whereas phages exhibit a high specificity for restricted sub-populations making the treatment with phages safer 10.

Long-Term Use of Phages is Safer than Antibiotics

As bacterial pathogens evolve to fit their environmental niche, phages adapt to the development of bacteria (11). The behavior of bacteria to develop resistance against phages has been studied well. The research of several groups indicates that creating a “Phage Crisis” similar to an “Antibiotics Crisis” is highly unlikely 12.

Evading Contamination by Other Phages during Purification

The biggest problem for native or environment-derived phages is that during isolation prophages or other phages can be isolated as well. As we plan on using only sequenced DNA of phages for our assembly, there is no possibility that we unintentionally generate a mix of phages.

Pathogen-Free Production of Therapeutic Phages

The major advantage of our method is that we have proven to produce clinically relevant phages for life-threatening pathogens. In contrast to conventional production methods, there is no need to cultivate these pathogens for Phactory. This feature was demonstrated with the production of phages against enterohemorrhagic E. coli (EHEC) phages in our low biosafety-level laboratory, whereas the EHEC bacteria themselves require biosafety level 3, similar to Salmonella typhi oder Yersinia pestis13.

Phages for the industrial sector

The application of phages is not only restricted for a clinical practice. It has also been evaluated for the agricaltural sector as a replacement of antibiotics. The administration was easy, successful and no side effects were seen14.

Overall Effect on Public Health

As clearly presented in the previous sections, phage therapy is a safe and promising treatment option, reducing side effects by high specificity15. We demonstrated that phage production is easy, stable, economic and safe, creating an ideal treatment for industrialized but also developing countries. One of the biggest problems we discovered with our survey was the public awareness regarding the use of antibiotics and the knowledge about other treatment options. This indicates, that the people have to be further educated about the risks of needless use of antibiotics. Moreover, the practical application must be further supported and tested as we did in the course of our iGEM project.

References

  1. Chan, Margaret. "Antimicrobial resistance in the European Union and the world." World Health Organization (2012).
  2. Chan, M. "Combat drug resistance: no action today means no cure tomorrow." World Health Day 6 (2011).
  3. Hughes, James M. "Preserving the lifesaving power of antimicrobial agents." Jama 305.10 (2011): 1027-1028.
  4. Lozupone, C. A., Stombaugh, J. I., Gordon, J. I., Jansson, J. K., Knight, R., Diversity, stability and resilience of the human gut microbiota. Nature 2012, 489, 220–230.
  5. Expert round table on acceptance and re‐implementation of bacteriophage therapy. "Silk route to the acceptance and re‐implementation of bacteriophage therapy." Biotechnology Journal 11.5 (2016): 595-600.
  6. Lin, Derek M., Britt Koskella, and Henry C. Lin. "Phage therapy: an alternative to antibiotics in the age of multi-drug resistance." World journal of gastrointestinal pharmacology and therapeutics8.3 (2017): 162.
  7. Slopek, Stefan, et al. "Results of bacteriophage treatment of suppurative bacterial infections. VI. Analysis of treatment of suppurative staphylococcal infections." Archivum immunologiae et therapiae experimentalis 33.2 (1985): 261-273.
  8. Fridkin, Scott K., and Robert P. Gaynes. "Antimicrobial resistance in intensive care units." Clinics in Chest Medicine 20.2 (1999): 303-316.
  9. MCCURDY, ROBERT S., and ERWIN NETER. "Effects of penicillin and broad-spectrum antibiotics on the emergence of a gram-negative bacillary flora in the upper respiratory tract of infants." Pediatrics 9.5 (1952): 572-576.
  10. Frisch, Arthur W., and Philip Levine. "Specificity of the Multiplication of Bacteriophage." The Journal of Immunology 30.1 (1936): 89-108.
  11. Lenski, Richard E., and Bruce R. Levin. "Constraints on the coevolution of bacteria and virulent phage: a model, some experiments, and predictions for natural communities." The American Naturalist 125.4 (1985): 585-602.
  12. Örmälä, Anni-Maria, and Matti Jalasvuori. "Phage therapy: should bacterial resistance to phages be a concern, even in the long run?." Bacteriophage 3.1 (2013): e24219.
  13. Chosewood, L. Casey. Biosafety in microbiological and biomedical laboratories. Diane Publishing, 2007.
  14. Colom, Joan, et al. "Microencapsulation with alginate/CaCO 3: A strategy for improved phage therapy." Scientific reports 7 (2017): 41441.
  15. Golkar, Zhabiz, Omar Bagasra, and Donald Gene Pace. "Bacteriophage therapy: a potential solution for the antibiotic resistance crisis." The Journal of Infection in Developing Countries 8.02 (2014): 129-136.