Human Practices | Heidelberg - iGEM 2022

Human Practices

Introduction

„Knowledge is power“ - a very famous quote by the English philosopher Francis Bacon. Our human practices team set the goal to spread this power throughout society. We wanted to raise the interest in science of people outside the academic field, as well as spread awareness of the dangers caused by the herpes simplex virus.

The herpes simplex virus can cause viral encephalitis in severe cases. Right now there are neither specific therapeutics against the herpes simplex virus on the market nor is there a preventing vaccine. The current therapy is based on a synthetic nucleoside analogue - acyclovir. Due to the nephrotoxicity and the high amounts of administered acyclovir needed for a successful treatment, we devised a safer and more target-oriented way to help people suffering from viral encephalitis by applying the treatment via the nose-to-brain route.

In the beginning of our project finding phase, we realised that even though every single one of us is a student with a scientific background, we ourselves weren’t aware of the risks resulting from a herpes simplex infection.

Therefore, we decided to structure our work in two parts: Collecting information and sharing information.

Collecting Information

As students know, a good professor must not know everything, but must be willing to learn about the topics he/she isn’t familiar with and pass it on to the students. We tried to imitate this as the human practices team by interviewing the people who know things we want to learn more about.

This strategy didn’t only apply to experts in the field of virology and immunology, but also in the field of science communication, education, industry and patent law. At the same time, we tried to get insights through a poll directed at our stakeholders, namely people suffering from a herpes simplex infection.

Only by undergoing this process, we were able to modify our project so that we could bring it to its full potential.

Sharing Information

As mentioned before, even some of us didn’t know about the herpes simplex virus being capable of causing neuronal damage. How can one introduce a new pharmaceutical to the market when people are not even aware of the seriousness of the disease? To spread this awareness, we thought of different events to share with the community. During our first attempts of explaining the topic and our project to strangers as well as to our sponsors, we noticed that people without scientific background cannot be thrown into the depths of virology that easily. Therefore, we decided to start with a simpler approach – the basics. What is DNA? How can we extract it? Do you know this scary virus dominating the world since 2020 – yes? Well, there are other viruses too and they can be just as dangerous.

We wanted to offer information online as well as in person and reach as many people with different backgrounds as possible. This is why we used different formats such as holding a lesson both in primary and in a community school, organising a science booth in the old town of Heidelberg, and creating an online quiz on the herpes simplex virus.

At the end of the day, we hope that we thereby managed to increase the knowledge and power of the people we met as well as ours. We can definitely state that exchanging information is a win-win situation, because people ask questions and questions promote thinking which helps us to obtain fresh perspectives and consider new aspects.

Interview with Prof. Haefeli

In the beginning of our project, when we as the new iGEM Team Heidelberg 2022 were brainstorming in which direction we wanted to take our project and what we wanted to achieve within the given year, we soon realised we all shared a common passion for the design and the production of small biomolecules, which make a great impact on the human biology, in our case the human health. This also falls in line with the general dedication towards medical sciences within the research facilities of Heidelberg, meaning that we would also have access to many resources and highly professional consultation on our University Campus for our medically-pharmaceutically oriented iGEM project.

When looking at different diseases to which to date only insufficient treatment or therapies with major drawbacks exist, a particular condition quickly captured our interest: Viral encephalitis, the most common type of brain inflammation, affecting nearly 1,5 million people globally per year and severely damaging the human brain, which leads either to death or in most cases to many disability-adjusted life years (DALYs) (Wang et al., 2022). These encephalitides, such as herpes simplex encephalitis, rabies, or tick borne encephalitis, are difficult to treat, because the brain is protected by the blood-brain barrier, which acts as a tight selective filter and blocks most drugs from entering the brain and exerting their effect in the affected regions (Patabendige et al., 2014). Because the brain is one of the most important organs of the human body, a high specificity of the drug is furthermore required in order to minimise ny possible side effects. This makes drug development for viral encephalitides extremely difficult and presents a challenge that remains to be addressed in the field of pharmaceutics,which we set out to address in our iGEM project.

Upon further research into viral encephalitides around the world, it soon became clear that there is a wide variety of viruses that cause different encephalitides. This made us realise it would be even better to not only develop a drug merely targeting one single type of virus, but rather a platform with which, once an effective route of drug delivery is established, one can readily target many different viral infections in the brain. We believe we have found a perfect candidate for this type of platform in siRNA; a class of biomolecules that gets utilised by intracellular defence mechanisms to degrade any foreign intracellular RNA with a complementary sequence to the siRNA (Elbashir et al., 2014). We also found a way to produce siRNA with a plasmid transformed into an E.coli bacterium (Huang & Liebermann, 2013), which means that by simply changing an insert sequence in our plasmid, we can effectively target and degrade many different mRNAs for proteins essential to the assembly of the respective virus, which would stop the virus from spreading even further. Lastly, siRNA is a fully biodegradable and biocompatible molecule, making this drug a promising safe candidate for application in humans.

As our method of drug delivery, we chose a recent new route of transporting small molecules to the brain: The nose-to-brain route, describing the mechanism of vesicles such as liposomes being transported retrogradely along both the olfactory as well as the trigeminal neurons into the brain, where the enveloped drug then exerts its effect (Lochhead et al., 2015). This avoids the great problem of having to pass the blood-brain barrier and guarantees a more effective drug delivery into the brain, making liposomes a technology we wanted to utilise for our drug against viral encephalitides.

For our iGEM project this year, we had to choose a specific type of viral encephalitis against which we could prove the effectiveness of a drug created by our platform. One candidate was the herpes simplex encephalitis (HSE), one of the most prevalent types and the deadliest in the western world, against which moreover no vaccine exists to date. Also being citizens of the western world, we wanted to learn more about this condition caused by the herpes simplex virus (HSV). Therefore, we organised an interview with Prof. Walter E. Haefeli of Heidelberg University Hospital, who is heading the Department of Clinical Pharmacology and Pharmacoepidemiology and is currently also involved in a clinical trial evaluating antibody-based treatment of HSV I and II.

This first thing we learnt is that HSV infections are quite persistent. Once the virus has for example taken up residence in the trigeminal ganglion, it remains there for life and poses a constant risk of breaking out again in response to environmental stimuli. These outbreaks in neurological areas cause strong pain, especially if the herpes virus reaches the optical nerve. Once it reaches the brain, the inflammation can be fatal, making novel effective drugs for HSE urgently needed and research in this area essential.

We also learned that while around 90% of the world’s population is infected with HSV (subtypes I and II combined), HSE is most dangerous in newborns and immunocompromised patients, where it can quickly lead to severe consequences. This also poses a great problem when looking at the current therapy, acyclovir, a nucleoside analogon which inhibits DNA replication of the virus inside its host cell. Because it is an antimetabolite and therefore has a very short half-life, it has to be administered very regularly (up to 5 times a day) - but at the same time, its metabolite is also quite nephrotoxic, which means that this treatment can strongly damage the kidneys and can often lead to acute renal failure (Yildiz et al., 2013). In addition, many strains are already resistant to acyclovir, and these strains are more likely to develop in immunocompromised patients, meaning that this already highly vulnerable patient group is even less likely to be successfully treated with acyclovir.

We presume that this would also be far less of a problem in our drug, because we produce the siRNAs in such a way that many siRNAs with a variety of sequences are produced in and extracted from E.coli. This will likely reduce the possibility of resistant mutants arising drastically, because these would have to either mutate in every single targeted sequence to such a degree that the siRNA does not complementary bind to it any more, or produce a completely new powerful protein interfering with the RISC complex. Therefore, our drug is also very likely to be a novel and better alternative for immunocompromised patients, one of our key stakeholders.

In conclusion, following this interview, we as a team were convinced to set our goal in developing this very much needed new drug against HSE, and that we would be able to help many people all around the world with the final product of our project.

VAAM Annual Conference 2022

Interview with Anja Störiko from VAAM

The VAAM (Association for General and Applied Microbiology) aims to implement research in microbiology for the benefit of society and the environment. For this reason, it promotes the exchange of information and cooperation among its members. At the same time, it acts as a point of contact for questions from the public (VAAM - Vereinigung Für Allgemeine Und Angewandte Mikrobiologie / VAAM - Vereinigung Für Allgemeine Und Angewandte Mikrobiologie e.V., n.d.).VAAM is also involved in publishing articles from iGEM teams in the journal Biospektrum.

On February 21st, 2022 Anja Störiko, responsible for editorial support for Biospektrum at the VAAM, conducted an interview with award-winning teams of the 2021 iGEM competition. Amongst them were team members from Bochum, Darmstadt, Heidelberg, and Marburg. The interview occurred online during the 2022 VAAM Annual Conference on February 21st, 2022. The interview can be seen online at: https://www.youtube.com/watch?v=n1-mfjeztgY

After briefly introducing our 2021 project, Ms. Störiko asked what organizational lessons we would like to integrate from last year into this year's project. In doing so, Jonathan, one of our advisors this year, cited, among other things, that this year we paid particular attention to the broad diversification of majors among team members. We also wanted to identify experts and stakeholders for our research project more timely and incorporate their feedback into our project. Regarding the question related to how our project idea this year came about, Ms. Störiko was particularly enthusiastic that the idea came directly from us students and was not simply given by a professor or Ph.D. For this very reason, it is also essential that we obtain evaluations and advice from people who already have experience in the respective fields.

At the end of the interview, each team was also asked whether and how their work would be continued after the competition. In this regard, our team mentioned the possibility of continuing the work as a bachelor's or master's thesis. At the same time, other groups can follow up on our project in the following years and further elaborate the research approach. The competition already encourages this opportunity through the use of publicly available Biobricks, which are continually used by teams from previous projects and added to for future groups.

What our team found most positive about the competition was the fact that you can work independently on a project that interests you. At the same time, you also come into contact with all the areas you would hardly hear about during an internship in a workgroup, such as financing. Thus, the competition not only encourages the individual participants to learn new methods but also allows them to determine which areas a research project ultimately depends on and in which one they would like to work in the future.

In summary, this interview increased the outreach of our team and our project. At the same time, it allowed us to reflect once again on what insights we want to incorporate into this year's project work from last year, how our project can be developed further so that society can benefit from it in the future, and what each participant will take away from the project work for his or her own future career.

You can get a detailed overview over all our Human Practices efforts on these following sub-pages:

REFERENCES

  • Elbashir, S. M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K., Tuschl, T. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498.
  • Huang, L., Lieberman, J. (2013) Production of highly potent recombinant siRNAs in Escherichia coli. Nat Protoc 8, 2325–2336.
  • Lochhead, J. J., Wolak, D. J., Pizzo, M.E., Thorne, R. G. (2015) Rapid Transport within Cerebral Perivascular Spaces Underlies Widespread Tracer Distribution in the Brain after Intranasal Administration. Journal of Cerebral Blood Flow & Metabolism, 35(3):371-381.
  • Patabendige, A., Defres, S., Keller, S., Vidyasagar, R., Michael, B., Das, K., Roberts, N., Parkes, L., Craig, A., & Solomon, T. (2014) The BBB in Encephalitis: Inflammation & a Role for Steroids? Journal of Neurology, Neurosurgery & Psychiatry, 85(10), 124.
  • VAAM - Vereinigung für Allgemeine und Angewandte Mikrobiologie / VAAM - Vereinigung für Allgemeine und Angewandte Mikrobiologie e.V. (n.d.). Retrieved October 13, 2022, from https://vaam.de/.
  • Wald, A., Corey, L. (2007) Persistence in the population: epidemiology, transmission. In: Arvin A, Campadelli-Fiume G, Mocarski E, et al. (eds) Human Herpesviruses: Biology, Therapy, and Immunoprophylaxis. Cambridge: Cambridge University Press, Chapter 36.
  • Wang, H., Zhao, S., Wang, S., Zheng, Y., Wang, S., Chen, H., Pang, J., Ma, J., Yang, X., & Chen, Y. (2022) Global magnitude of encephalitis burden and its evolving pattern over the past 30 years. Journal of Infection, 84(6), 777-787.
  • Yildiz, C., Ozsurekci, Y., Gucer, S., Cengiz, A. B., & Topaloglu, R. (2013). Acute kidney injury due to acyclovir. CEN case reports, 2(1), 38–40.