Expert Opinions
We had identified the problem and created a solution, but we still had many unanswered questions that we could not find answers to in the literature. Therefore, we consulted with physicians and researchers for answers but also their opinions on NiSkin. We did this from the beginning and continued this throughout our project to guide our project and ensure that we had time to change it if necessary.
Which Skin-Penetrating Peptide To Use?
Our initial project did not include a skin-penetrating peptide because we did not consider the basement membrane or the other layers of the skin that form a barrier to topical treatments. We needed to identify a skin-penetrating peptide and assure its efficacy for our project.
Dr. Stefano Menegatti
Associate Professor at North Carolina State University in the Department of Chemical and Biomolecular Engineering
Dr. Menegatti wrote a paper titled “Peptides as skin penetration enhancers: mechanisms of action," which discussed several transdermal peptides. He told us that TD-1 is the best transdermal peptide to use for our project, told us that he believed our project was feasible. He also made us consider the optimal concentration of nisin in the skin and whether it can be toxic at high levels.
Will TD-1 Transport Nisin?
Once we confirmed that we will use TD-1, we needed to figure out if TD-1 will transfer nisin considering its size and structure.
Dr. Xin Guo
Associate Professor at Shandong University, China in the Department of Public Health
Once we established that we were going to use TD-1, we still had questions on its mechanism of action and whether it can transport a molecule the size of nisin through the skin. So, we contacted Dr. Guo, who wrote papers on TD-1. She recommended that we talk to Dr. Renquan Ruan.
Dr. Renquan Ruan
Founder of MELLGEN BIOTECH, a startup focusing on recombinant transdermal peptides
Dr. Ruan confirmed to us that molecules with nisin’s size can be transported through the skin with TD-1.
Is There An Attraction/Need For Our Project?
At this point, our project involved creating a fusion protein with nisin and TD-1 to reach the dermis. To see what they think about our project, side effects, topical treatments in general, and antibiotic resistance, we consulted physicians.
Dr. Mark A. Russell
Dermatologist and Professor of Dermatology, Otolaryology, and Pathology at the University of Virginia School of Medicine
Dr. Russell told us that “the use of topic treatments for skin infection is attractive to reduce bacterial resistance to antibiotics and I would consider using it if it was cost-effective, had a favorable risk/benefit ratio, and had an acceptable side effect profile.” This ensured us that side effects are major considerations/concern.
Dr. Molly Hughes
Infectious Disease Physican, Professor, and Researcher at the University of Virginia investigating biodefense and emerging pathogens.
Dr. Molly Hughes told us that current cellulitis treatments are oral and IV treatments. She told us the price ranges, how long treatments usually last, side effects, and about antibiotic resistance. She also informed us that immunocompromised patients have to take treatments for longer periods of time and have more severe side effects. She recommended that we do a combination therapy with our product and IV/oral treatments.
Dr. Richard Flowers
Dermatologist and Assistant Professor at the University of Virginia.
Dr. Richard Flowers found the appeal of this project, saying that it is attractive. He also told us about antibiotic resistance with current treatments of cellulitis, such as doxycycline. The examples that he provided on antibiotic resistance stressed how big of an issue antibiotic resistance is and how NiSkin is attractive because of this.
Dr. Sifri D. Costi
Infectious Disease Physician, Professor, and Researcher at the University of Virginia investigating hospital epidemiology, immunocompromised infectious diseases, molecular pathogenesis and epidemiology, innate immunity
Dr. Costi informed us about the history of antibiotic resistance and how NiSkin is attractive because it means one more option to use in case antibiotic resistance to another cellulitis treatment arises.
From our meetings with all of these physicians, we learned that there are many current examples of antibiotic resistance against cellulitis treatments, side effects are something to consider when choosing medications, and that there are no current topical treatments in the United States that are as effective in treating cellulitis as oral and intravenous treatments.
How Does NiSkin Compare To Topical Treatments Used In Other Countries?
When we were discussing NiSkin to physicians, we were talking to physicians in the United States. We soon found out that there is a topical treatment for cellulitis in other countries, but not the United States, that reaches the dermis, and it is called ‘Fucidin.’ We had to understand its mechanism of action and how NiSkin differs from it. So, we had to talk to a physician who worked with Fucidin.
Dr. Greg German
Phage and Chronic Infection Clinical Physician at St. Joseph’s Hospital and Assistant Professor in Department of Laboratory Medicine and Pathobiology, University of Toronto.
Dr. German told us that Fucidin goes through the skin in the presence of an abscess, which creates an opening. So, unlike NiSkin, Fucidin requires an opening in the skin and cannot simply penetrate through the skin.
What/How Should We Model and Experiment?
We had begun modeling earlier during the project as we had many questions based on the literature that we could not find answers to in the literature. Before we could actually do experiments, we had spent time making and revising protocols. Some of the protocols that we made were inspired or from other protocols published, and we contacted some of the authors for clarification.
Dr. Anders Nelson
Postdoctoral fellow. Doctorate of Pharmacology, University of Virginia.
Dr. Nelson suggested possible modeling projects to try and explained how clinical questions are valuable and can be answered with modeling. Dr. Nelson also confirmed our approach to modeling diffusion equations.
Anton Du Preez Van Staden
CEO of FluoroBiotech, focusing on developing recombinant proteins using E. coli technologies.
Dr. Anton Du Preez Van Staden helped our team with our NisA purification protocol, specifically with how to lyse our cells.
Dr. Marcos Pires
Associate Professor of Chemistry and Director of Graduate Studies, University of Virginia.
Dr. Pires talked to our team about the T7 promoter system and experiment optimization. He recommended that we test a variety of protein induction conditions, which led to us optimizing IPTG concentration, temperature, and induction period.
Dr. Brent A. French
Professor and Researcher and Department Chair of BME, University of Virginia.
Dr. French discussed the V8 protease and effects on our product. Dr. French influenced us to use the V8 site.
Public Opinion
We knew what experts and potential prescribers thought about our project, but we also wanted to know what the public thought about different modes of treating skin infections, antibiotic resistance, and side effects to see what they would think about our project.
To see what the public thought about our project, we released a 9-question survey with questions allowing us to know which modes of treatment people are most comfortable with, which side effects people are least comfortable with, and whether people are, in fact, concerned with antibiotic resistance.
We first had to get our survey approved by the IRB-SBS, which is the Institutional Review Board for the Social and Behavioral Sciences at the University of Virginia. Our team had to do training about research practices and confidentiality. We also had to create a protocol explaining what our survey was, who are participants will be, what experience or knowledge each member of our team as well as our lead advisor have with working productively and respectfully with participants, how we would distribute the survey, how we will receive consent, how our data will be stored and deleted, how people can withdraw, and the risks and benefits of our survey. Once our protocol got approved (protocol #6001) by the IRB-SBS, we were able to release our survey to people outside of the university.
Survey
Survey Questions
Survey Responses
This histogram displays the survey responses to the questions asking about people’s comfortability to different modes of treating skin infections. A ranking of 1 means that someone is not comfortable with the treatment, and a ranking of 5 means that someone is comfortable with the treatment. As can be seen in the graph, people are typically most comfortable treating skin infections with oral or topical treatments. Not only that, but there is no significant difference between how comfortable people feel between oral and topical treatments; they are equally comfortable with both. People are also not as comfortable with intravenous treatments, which is a common treatment for cellulitis.
One of the aspects of NiSkin rendering it unique is the fact that it is a topical treatment. We wanted to know if people wanted a topical treatment for a skin infection in the first place. When we were still beginning our project and interviewing experts, we saw the trends explained earlier in the survey responses. This showed that there is a need for NiSkin as people would appreciate a topical treatment since they are equally comfortable between topical and oral treatments. We looked at the survey results throughout the project and there were the same trends as shown in the graph above, again highlighting the appeal for NiSkin.
In summary, these survey results highlighted to us that people were most comfortable with topical and oral treatments for treating skin infections, and there was no significant difference between comfortability between the two methods. This meant that NiSkin being a topical treatment would be attractive to the public.
This graph depicts the level of concern that people have to different side effects of oral treatments. The survey did not tell people that these side effects had to do with oral treatments, however. A ranking of 1 means minimal concern whereas a ranking of 10 means that people are extremely concerned. The trend here is that people mostly were highly concerned with these side effects, ranking from 7 to 10.
A big part about NiSkin is increasing comfortability to cellulitis patients. So, we wanted to know if people actually cared about side effects. Evidently, they do, given these results. When we were in the beginning stages of our project and saw these results, we knew that it was correct of us to take side effects into account since people do in fact have concerns about them. As we did lab work and completed lab work, we continued to see this trend and were further ensured that side effects are in fact a serious concern, making a topical treatment like NiSkin necessary.
In summary, these survey results showed us that the public was highly concerned with side effects that are characteristics of oral treatments. This meant that our team’s goal to prevent these side effects is necessary.
This graph displays people’s levels of concern with antibiotic resistance. A ranking of 1 means little to no concern with antibiotic resistance, while a ranking of 10 means an extremely high level of concern. As can be seen in the graph above, people are mostly highly concerned with antibiotic resistance, ranking higher than 5.
Another key element of NiSkin is the use of nisin, which MRSA has not gained full resistance to. So, we discovered another treatment to use against antibiotic resistance bacteria like MRSA. We wanted to know if this was something that greatly appealed to people and if we should worry about MRSA and antibiotic resistance. Given these responses, we knew that NiSkin is appealing then because of this nisin feature. We saw this similar trend of high concern throughout the journey of NiSkin, continuously highlighting to us that people do in fact worry about antibiotic resistance.
In summary, these results showed us that the public were mostly highly concerned with antibiotic resistance. This also meant that our decision to provide another option to combat antibiotic resistance is both necessary and attractive.
Collaboration
As we were working on our project, we looked at what projects the other iGEM teams were working on to see if there was a team working on a project similar to ours and we could work with.
Instituto Tecnológico de Chihuahua
During our meeting with the Tec-Chihuahua team, we were asked questions about degradation, Nisin’s half-life, and temperature for storage. These questions made us realize that something that we need to start considering is storage standards for our product such as what temperatures it can be stored in.
Mid-Atlantic Meetup
This year, our team also hosted the Mid-Atlantic Meetup. The Mid-Atlantic Meetup is an event where iGEM teams in the Mid-Atlantic Region present their projects to other teams and receive feedback. This year we hosted William & Mary, University of Maryland, East Coast Bio Crew, University of North Carolina-Chapel Hill, and North Carolina State. When our team presented NiSkin, we received questions on the body’s response to nisin. This led to us doing more research and confirming that nisin is relatively harmless as it is a peptide and does not have an antibody, so T-cells cannot bind to it.
How Did Opinions Guide Our Project?
In terms of expert opinions, our interviews allowed us to know which skin-penetrating peptide to use: TD-1. It gave us the assurance that TD-1 will be able to transport NiSkin to the dermis.
Talking to physicians educated us further on current methods of treatments for cellulitis, their side effects, and why there are no topical treatments used. It also introduced the unfortunately high number of cellulitis treatments that no longer are used because of antibiotic resistance, and it gave us more detail on the history of antibiotic resistance. Physicians also helped us understand what is taken into account when being introduced a new treatment, the importance of comfortability to patients, and the appeal for topical treatments.
By talking to experts and physicians, we were guided to do more research on antibiotic resistance to understand the gravity of the situation for cellulitis treatments. It also guided us to consider how we frame our problem statement and explain if we are focusing on Group A Strep as well. Lastly, it guided us to do more research to see how exactly NiSkin differs from Fucidin.
For public opinion, the results that we saw in our survey guided our project by showing us that we should continue our project in the path that we are on, which involves making a topical treatment, reducing side effects caused by oral treatments, and finding a way to combat antibiotic resistance.