Over the course of the 2023 iGEM season, the UChicago Genehackers engaged in integrated human practices that enhanced our understanding of the applications and impact of our project. The human practice-related activities were divided into three distinct categories. First, we corresponded with academics who published the core papers we used to inform our decisions relating to the synthesis of levothyroxine and biocatalysis using transaminases. Second we increased our understanding of the current iGEM competition landscape through collaboration with the University of Michigan iGEM team culminating in a global iGEM biomedical health equity symposium. Finally, we fostered lasting connections on the southside of Chicago and on campus by bringing our synthetic biology experience to demonstrations in elementary and middle schools.
Input from leaders in academia
To effectively tackle a scientific problem with real-world applications, it is essential to understand the literature which forms the ground on which the project methodology stands. Throughout the summer leading up to the iGEM conference, the UChicago Genehackers conducted correspondence with academic leaders in transaminase mediated-synthesis methods and total synthesis of our drug target levothyroxine. The results of these messages informed our choices about targets, substrates, transaminase variants, and model organism strains.
Professor Rosario Brieva Collado
Coauthor of Amine Transaminase Mediated Synthesis of Optically Pure Piperazinones and 1,4-Diazepanones
We reached out to Dr. Brieva Collado to gain more information about using transaminases to synthesize drugs which are optically pure. Current levothyroxine synthesis methods require a racemate clarification which discards half of potential product. The benefit of a transaminase is that all of the drug produced is optically pure. Dr. Brieva Collado believed that the transaminases she used were not appropriate for the synthesis of levothyroxine. Due to her input we were able to narrow down the transaminases that we considered for our synthesis. She also suggested that we reach out to Dr. Wolfgang Kroutil to learn more about the substrates and amine donors involved in our anticipated synthesis project.
Professor Wolfgang Kroutil
Coauthor of
⍵-Transaminases for the synthesis of non-racemic ⍺-chiral primary
amines
Author of
Asymmetric Preparation of prim-, sec-, and tert-Amines Employing
Selected Biocatalysts
Following our discussion with Dr. Brieva Collado we next reached out to Dr. Kroutil to learn about the feasibility of using our substrate given the transaminases we were now considering. Dr. Kroutil gave us suggestions about a few different ways to make the chiral center of an L-alpha amino acid. He suggested that we start with an alpha-keto acid as a precursor and warned us that the size of the substrate might be a challenge. Based on this advice we chose to synthesize a smaller fragment of the larger drug which we termed “minithyroxine”. This levothyroxine fragment was designed in such a way that once produced it could easily be converted into the full scale drug.
iGEM Collaboration
In the summer of 2023, the UChicago Genehackers initiated a strong partnership with the University of Michigan’s iGEM team. Frequent cross-team meetings allowed us to come up with new and creative ways to engage with our community and use the global reach of iGEM to produce the best project that we could.
Framing our Project and Discussions on Next Steps
The UChicago and UMich iGEM team boards met several times throughout summer 2023 to share our projects and outline potential collaborations. The UMich project focused on creating a simplified diagnostic kit for a single nucleotide polymorphism found disproportionately in people of African descent. They suggested that we consider global health equity in the implementation of our project, which quickly became an essential part of our human practices work given the high global prevalence of hypothyroidism. Since our strategy focused primarily on promoting green chemistry and reducing environmental waste from synthesis, we helped UMich to consider an environmental angle on their own project. Additionally, we discussed potential wet lab and dry lab collaborations. This fruitful conversation led us to make a commitment to reciprocally confirming results via peer review in the 2024 iGEM cycle.
Drylab Collaboration
The UChicago and UMich IGEM teams met in the early fall to discuss drylab improvements and potential future collaborations. They introduced us to the Haddock software for protein-protein interactions in MD systems, which can also be used to describe the energetic properties of the protein, like binding free energy and flexibility. We provided useful details on our modeling of protein-substrate interactions, such as a transaminase with PLP and its amine donors/acceptors. Based on this meeting, we gained a clearer understanding of which software is most effective for MD simulations and what kinds of interactions and structures we can feasibly simulate. Crucially, we discussed the critical role of drylab in IGEM and how to better integrate computational work into our current and future projects.
Discussing Health Equity
The UChicago and UMich iGEM teams are planning a joint podcast for the University of Michigan's collaborative iGEM podcast series. During this podcast, we will introduce our projects and discuss their potential applications in global health equity. We also plan to discuss the broad range of projects that can contribute to global health equity in advance of choosing our teams' projects for the iGEM 2024 year. Projects that improve global health can include simplified diagnostics, enzymatic biocatalysis, improved drug delivery, commercially available scientific kits, and cheap disease monitoring devices.
Attending Global Biomedical Health Equity Symposium
We attended the Global Biomedical Health Equity Symposium on October 1, 2023, which featured 7 iGEM teams from around the world, including the host UMich. Our presentation focused on the ability of enzymatic synthesis to improve accessibility for the highly-prescribed hypothyroidism drug levothyroxine. Hypothyroidism affects up to 5% of the population and is a chronic illness that can be treated effectively with a daily levothyroxine dose. However, the cost of levothyroxine is prohibitive in some developing countries, while in others limited levothyroxine supply means that few receive treatment. Additionally, levothyroxine is a narrow therapeutic index drug, meaning that small dose deviations can lead to adverse effects. In developed countries, companies use a complex and expensive purifying process to remove bioactive impurities generated during synthesis. Using the GLOW method, implementing our biocatalytic method in place of heavy chemical synthesis can improve levothyroxine quality and price accessibility. Transaminases are very substrate-selective and stereospecific, meaning that companies will not have to throw out the 50% of the product that is the wrong enantiomer and that levothyroxine will be produced with fewer impurities. This can ensure that more people have access to high-quality levothyroxine at a lower price.
Community Outreach and Organization
Inspiring Researchers on Campus
In the spring of 2023, the UChicago Genehackers embarked on an effort to highlight the exciting undergraduate research that takes place on our campus. In the first annual Genehackers Research Symposium, undergraduate researchers from divisions as diverse as particle physics, tissue engineering, chemical biology, and geological sciences presented to their peers in a constructive yet rigorous event. The well-attended symposium allowed for undergraduates who have not yet committed to research labs to hear about what was available, while those who were actively involved in labs across campus could gain new perspectives and generate ideas for future work!
Bringing Science to Underserved Communities
In the winter of 2023, the UChicago Genehackers partnered with the Bret Harte Magnet Cluster school on the southside of Chicago to bring synthetic biology demonstrations into the neighborhoods around our campus. Engaging with elementary school students, representatives from our team taught lessons about miscibility and solubility before leading an interactive demonstration where students experimented with water, oil, corn starch, food dye, and soap before building their own mini lava lamp with products they can find around the house. Engaging with middle school students, representatives from our team taught lessons about DNA and chromosomes. Working together, students learned to extract DNA from strawberries and to understand what its function within the organism.