In order to enhance feedback collection for the project, we intend to gather real-time challenges encountered by both the experimental and modeling groups, while also seeking input from authoritative experts in relevant fields. Our objective is to identify effective solutions that will further advance the progress of the project.
Questions
Nanjing Normal University:
Following the confirmation of the project's design concept, the students in the experimental group commenced their experiment. However, they encountered several challenges during the process, including determining optimal drug delivery methods (whether to employ fixed-point blasting or upon entry into the human body). Simultaneously, the students in the modeling group also faced difficulties while constructing a molecular docking model. Consequently, we sought assistance from two esteemed experts who possess extensive expertise in these domains - Professor Xiujuan Li and Professor Chao Ye(Figure.1).
Nanjing Tech University:
The iGEM team project of Nanjing Tech University employs probiotics to manipulate the intestinal environment and modulate intestinal inflammation, which is highly pertinent to our own project. Consequently, we anticipate gaining valuable insights for our project through extensive communication. The team members and Professor Guannan Liu have offered suggestions on addressing issues and refining modeling ideas for our project pathway(Figure 2).
Nanjing Normal University
Experimental part:Professor Xiujuan Li provided numerous suggestions and ideas for the experimental group. Firstly, she affirmed and encouraged our ideas and the entire project but raised questions and suggestions regarding the expected goals and framework of our project.
1.Our aim was to achieve expression of multiple peptides in Zymomonas mobilis, as well as their entry into the human intestinal tract for colonization or targeted release. However, Professor Li suggested that we simplify our design and reduce our expected goals. She proposed that we first focus on achieving stable peptide expression in Zymomonas mobilis and demonstrate their ability to reduce glucose levels in a simulated human fluid environment in vitro. It is advisable not to involve biosafety and ethical issues such as mouse experiments or human trials due to long experiment cycles and various verification steps from oral intake to intestinal effects. Aspects related to colonization within the human body or other areas can be included as supplementary information or future directions in the project report.
2.Additionally, Professor Li advised us to further explore maximum tolerance data of such drugs within the human body, clarify bacterial expression indicators, determine optimal dosage of beneficial bacteria within the body, understand differences between drug administration versus bacterial administration including production processes, survival rates, metabolism patterns of strains used.
3.Finally, Professor also mentioned a website called "intestinal coating" which contains valuable information about potential food-grade delivery carriers like starch tablets or calcium alginate hydrogels that could be suitable for our research purposes.
Modeling part: Professor Ye Chao from Nanjing Normal University suggested that our modeling group should focus on a specific aspect due to the complexity, breadth, extensive scope, insufficient coherence, and limited time of the preliminary idea. He recommended commencing with protein interaction and excelling in protein prediction and molecular docking modeling. For the in vivo intestinal simulation model, he proposed considering in vitro experiments for verification. Additionally, he advised utilizing the efficient application Alphafold3 for structural prediction.
Harvest: The students in the experimental group found that the overall workload of the project was excessive during experimentation, resulting in less smooth progress than anticipated. They had concerns regarding project design and also encountered difficulties meeting the competition's requirement of abstaining from animal experiments. However, two professors from Nanjing Normal University suggested simplifying the design to make it more attainable. They also provided helpful solutions to overcome experimental challenges such as artificial intestinal fluid and intestinal coating.
Figure 1 Two professors from Nanjing Normal University
(Left: Professor Xiujuan Li, Right: Professor Chao Ye)
Figure 2 Photo taken at Nanjing Normal University
Nanjing University of Technology
Experimental part: The project at Nanjing University of Technology also focuses on probiotics, specifically utilizing Escherichia coli EcN as the material. The primary objective is to exert control over the intestinal environment and regulate intestinal inflammation. The team instructor has been highly supportive of our project.
1.Referring to the design and construction of the plasmid, they believe that we can leverage our understanding of the intestinal environment to develop a responsive system based on specific conditions within the gut microenvironment. It is crucial for us to determine whether the expression of our hypoglycemic or antihypertensive peptides would impact the intestinal microenvironment or other metabolic pathways.
2.The instructor reminded us that if we intend to express Zymomonas mobilis and deliver them to the intestine, it is essential to promptly apply for iGEM's safety whitelist in order to demonstrate that Zymomonas mobilis is a safe and effective probiotic (this application can be supported by FDA certification and comprehensive laboratory research on Zymomonas mobilis).
3.Additionally, during our study of Nanjing University of Technology's project, they introduced their design involving a lytic gene in ECN. When ECN concentration reaches high levels in the intestine, this lytic gene secretes lytic toxin which acts upon ECN cells, leading them to die off until reaching a certain concentration level. This gene effectively controls bacterial numbers within a safe range in the intestine. This concept has inspired us regarding suicide response design for our own project (Figure 3,4).
Modeling part: In response to the question of whether our team should dock DPP-4 with GLP-1 using the mutant GLP-1, the iGEM team of Nanjing Tech University suggested that we not only verify the ability of the mutant GLP-1 to prolong its half-life but also confirm its non-interference with GLP-1R interaction.
Harvest: After engaging in extensive communication with the team from Nanjing University of Technology, we acquired a more comprehensive understanding of our project's apoptotic response mechanism and explored the feasibility of utilizing glucose concentration as a triggering mechanism for our suicide switch. Furthermore, during this collaborative process, we identified another modeling concern that necessitates attention.
Figure 3 Communication with Nanjing University of Technology team
Figure 4 Communication with Nanjing University of Technology team
IGEM Team, Nanjing Medical University
We engaged in a comprehensive exchange of integrated human practices with the iGEM team at Nanjing Medical University.
1.In terms of publicity, they recommended utilizing videos for promotional purposes to enhance page views, which can be learned from. They suggested employing multiple platforms and accounts for publicity, such as TikTok, bilibili, and other renowned domestic and international film and television platforms.
2.Regarding education, they believe that organizing more roadshows within and outside the school would result in a broader coverage effect involving diverse groups of people. Concerning privacy concerns, they emphasized considering whether interviewees are willing to appear on camera or if their photos need to be coded to protect others' privacy rights—an issue our team should address.
3.During discussions about wiki , we deliberated on whether the promotional video should be homemade or outsourced but ultimately reached a consensus that it should be self-made and placed at the end of the wiki. Additionally, your team inquired about strategies for educational outreach and activity innovation; we provided relevant suggestions and answers accordingly.
Havest: Throughout our entire communication process, we maintained a spirit of mutual assistance through warm discussions where both teams openly shared their strengths without reservation—exemplifying effective communication's essence where brainstorming sessions fostered project promotion and best practices from both sides resulting in a win-win situation (Figure 5，6).
Figure 5 Communication with Nanjing Medical University team
Figure 6 Communication with Nanjing Medical University team