USAFA iGEM 2023 Lyme-AID

Documenting Our Journey

How we approached the Nine Human Practices Steps

Build a Diverse Team

• Gathered interest from a large community of students and educators
• Team open to students and faculty of all academic disciplines
• Team members chosen with different strengths and skillsets

Explore Context

• Each team member brainstormed a significant problem and proposed a solution
• The team investigated Sepsis and Lyme Disease concurrently, eventually electing to pursue the latter
• We spoke with AFRL researchers and local experts on Lyme Disease, and learned that our project could have big implications in the future of Lyme Disease detection

Brainstorm Broadly

• Within Lyme Disease detection, we researched multiple ways to create a biosensor patch
• We planned to speak with more experts and the public about the a potential rapid detection patch to make big advances in Lyme Disease detection

Document Progress

• After talking to experts, we took notes and tailored our research
• Following the inputs of others kept us on track and objective
• Speaking to others allowed us to get a sample of outside sentiment toward our ideas

Integrate Insights

• By speaking to the public and healthcare professionals, we learned that there are some concerns with our project
• We are actively taking steps to fix and prevent these concerns

Close the Loop

• More time, research, and development will be needed to take our project from where it is to where we want to be
• We will be working to keep Human Practices at the forefront of our future progress

Present Evidence

• The community’s positive disposition and eagerness for our ideas showed us the positive outcomes that could result from the advancement of our project
• A model organism will enable safe research and allow for progress against chronic vector-borne diseases
• A patch like ours is safe and ethical, as it doesn’t contain any live specimens

Connect and Share

• By producing results toward creating a model organism and therefore a biosensor patch, we enable others to expand upon their research
• By making these things available and known to the individuals and organizations that helped us along the way, our team has created meaningful relationships while contributing to disease research as a whole
• This will benefit many, even the individuals, institutions, and communities that we haven’t yet connected with

Carry it Forward

With the progress made in this project, other researchers could have a model organism for Borrelia burgdorferi with which they can conduct their own research. Team USAFA has the option to carry this project forward into 2024, and all our research and setbacks are available to others via our wiki page.

Putting it All Together

How we synthesized what we learned in terms of Reflection, Responsibility, and Responsiveness

Reflection: Our motivating values, system for prioritization, progression of goals, and understanding of the current field

Our project, Lyme-AID, was inspired by the ongoing disparities regarding timely and accurate diagnosis of tick-borne illnesses, namely Lyme Disease. These diseases have severe impacts that often increase in severity as time passes without a diagnosis. A large amount of these issues are derived from the tendency of Lyme Disease symptoms to mimic those of other common afflictions. Without proper diagnosis, symptoms will worsen, treatment will be delayed, and lives will be forever altered. Thus, moral values form the foundation of our project. After talking with numerous patients suffering from Lyme Disease, it was clear that the current lack in accessible diagnostic care was altering lives long-term. Our team was able to visualize the clear need for scientific advancement in this field and was determined to help stop lives from being negatively impacted.

There are many aspects of tick-borne illness that need to be addressed. In order to determine where to start, we organized a list of priorities. First, we needed to understand how these illnesses progress to decide where accurate and timely diagnosis fits in. Currently, accurate diagnosis is the first step in securing timely and thus effective treatment in Lyme Disease cases. Thus, we prioritized developing a system of diagnosis that would remedy the current issues, as timely diagnosis is crucial to maximizing effectiveness of current treatment methods.

Our goals for this project morphed over time as we became more aware of the reality regarding the status of Lyme Disease in the medical community. First, we aimed at creating a patch capable of both detecting the presence of Borrelia burgdorferi in the interstitial fluid, as well as indicating such bacterial presence to the untrained eye. However, as our team continued to conduct scientific literature reviews, communicate with experts in the field, and learn from current Lyme Disease patients and experts, we determined that the more pertinent first step was in detection. First, Borrelia burgdorferi is an organism that requires a BSL Level II Lab. We understand that not every lab has access to such facilities, so we aimed to create a model organism that could reflect the characteristics of Borrelia burgdorferi that we were looking to target for our detection method, but still be used in a BSL Level I Lab. To accomplish this, we sought to express characteristic surface proteins of Borrelia burgdorferi, OspA and OspC, on Escherichia coli. Following this successful expression, we aimed to determine which techniques could be used to actually detect these surface proteins, and our research led us to DNA aptamers with gold nanoparticles.

Our approach focuses on a diagnostic technique that can be applied by those without any medical or scientific training, and without any need to try and test the actual tick that bit them. Currently, the medical standard for diagnosis is via blood testing or PCR, both of which requiring time and access to sufficient medical care resources. Furthermore, Lyme Disease can be also identified by testing the actual tick that caused the bite within a laboratory, but again, this is contingent on ability to find, remove, and keep the specimen long enough to access a lab capable of testing. Other iGEM teams have used this approach to try and create more accessible systems of testing the ticks for Borrelia burgdorferi presence. While this avenue is important in the sense of understanding the components of the tick that indicate infection, it still had limitations in applicability. Specifically, it required the ability to find and remove the tick, while understanding how to crush it sufficiently to access the proteins indicative of Lyme Disease infection. Thus, our proposed approach differs in that we sought to test for specific proteins associated with Borrelia burgdorferi infection directly at the source of the bite.

Responsibility: Our intended use, accountability, and acknowledgement of the communities called in and out.

It is highly unlikely that the project could be misused. By engineering E. coli to express Salp12, the team bypasses modifying and testing Borrelia burgdorferi. This prevents researchers from being exposed to a dangerous pathogen and allows for the safe testing of a model Borrelia burgdorferi organism. The same principles apply in producing surface proteins and antigens for Borrelia burgdorferi in Escherichia coli.

By avoiding the use of higher BSL organisms in testing and research, and by attempting to produce a model organism for Borrelia Burgdorferi, USAFA iGEM has promoted safe and responsible research. Minimizing risk is an important part of safety, and being safe is a responsibility that all genetic engineers share. Advocacy for these concepts, possible with a functional model organism, allows our team to promote safety outside of our lab and inside the labs of other tick-borne illness researchers.

Any community invested in Lyme Disease treatment and prevention will be the most affected by our project. Individuals with the condition, their healthcare providers, researchers that create treatments and early detection measures, and individuals who want to keep themselves safe will be influenced by a potential model organism and microneedle patch which facilitates rapid and early detection of Lyme Disease.

There will be no negative impact if the project succeeds. The project is not intended to harm anyone nor place any individual or community at a disadvantage. However, it is possible that this research could be used for commercial gain. If made purchasable, individuals with less resources may have difficulty obtaining rapid detection microneedle patches. This possibility is far away due to progress that still needs to be made, and our team has chosen to work against this possibility by making our research and intentions public.

Responsiveness: Our inclusion of stakeholders, incorporation of feedback, and connection with the end goal

The most important community we consulted was comprised of those currently suffering from Lyme Disease. In understanding how accurate and timely diagnosis, or lack thereof, impacted their lives, we truly understood the importance of this research initiative. Also, since our proposed project is intented to benefit the medical community, the best way to understand what goals need to be prioritized was through talking to those who would have used it. We value the concept that Lyme – AID has the capability to play a direct role in people’s lives, and thus we were determined to understand the true needs of that proposed community.

Speaking to those who will realistically use this product in the future, or who could have used it in the past will provide valuable insight into the applicability of our proposed project. While research for the sake of advancing scientific knowledge is crucial, the diagnostic track is unique in that it demands consideration of other stakeholders – such as medical professionals and patients.

Our team can close the loop between our design and what is desired by ensuring the documentation of our processes. By clearly documenting the process we took to ascertain the background, demand, procedures, and end goals of our Lyme-AID process, we use our Human Practices work to inform our team’s ethical, technical, safety andcommunication decisions.