Overview
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In every phase of our project, from design to execution and final implementation, Integrated Human Practices served as a pivotal element. By incorporating input from experts, fellow iGEM participants, and the wider public, we've been able to effectively steer our project in the desired direction and set the stage for achieving our anticipated results.
Background
Our Own Story
In 2021, Litong Wu and Yanheng Zhu frequently played basketball games together. Litong Wu observed that his injuries, particularly to his joints, took longer to heal compared to Yanheng Zhu. Seeking the underlying reason, Litong realized his Body Mass Index (BMI)—a measure of body fat based on height and weight, where a value below 23.9 is considered normal—surpassed that of Yanheng's. More significantly, Litong was classified as obese, with a BMI exceeding 32. This revelation led to a question: is obesity negatively impacting his knee joints?
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Fig.1 Our student leader and student member played basketball together
Yingcai Program in China
Yingcai program is a national program to select a group of high school students with excellent character and learning ability to enter the university, participating in scientific research under the guidance of famous scientists.
In 2022, Wu Litong was selected for the Yingcai Program. With Professor Hua Zichun's team from Nanjing University, he researched the impact of a high-fat diet-induced obesity and adipose exosomes on post-traumatic osteoarthritis in young mice, particularly from DMM (destabilization of the medial meniscus) surgery. They also examined how obesity exacerbates the condition. Their findings suggested that obesity induced by a high-fat diet can worsen post-traumatic osteoarthritis resulting from DMM surgery in young mice. Furthermore, it was determined that obese adipose exosomes primarily impact the surface of the knee's articular cartilage.
In light of the previous research, we've embarked on an iGEM project with a new, exciting objective - how can we employ synthetic biology methods to devise innovative therapy forms that safeguard our joints from damage caused by obese adipocyte exosomes?
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Fig.2 The certification and the Poster in 2022 Yingcai biology program
Public Attitude
Nanjing-NFLS Alliance Meetup
We invited several PhD students from Nanjing University, University College London, University of Queensland, National University of Singapore, and University of California, San Francisco, and held a meeting online, in which we discussed our project and lab security.
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Fig.4 Nanjing-NFLS alliance Meetup with aboard universities
We received feedback about potential improvements and were alerted to possible risks associated with our product. Following this, The PhD students introduced the safety regulations governing lab operations to us. This comprehensive feedback and briefing greatly helped us refine our approach for future experiments.
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Fig.5 PhD students from aboard universities give us many useful advices on our project
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Fig.6 Nanjing-NFLS team exchanged safety and security rules with PhD students from aboard universities
The Attitude of Stakeholder in Pharmaceuticals
Novozymes Pharmaceuticals, established in 2012, is a global supplier of bioactive compounds and widely contributed in other fields as well. The company has been particularly noticeable for its indelible contribution to global efforts combating the new coronavirus, through the development and production of nucleic acids. Recently, we have had the chance to interview the Vice President and Technical Director of Novizen Pharmaceuticals, Mr. Feng Su, as well as Mr. Tang Bo, the Deputy General Manager.
Micro-RNA technology expertise: Our discussion primarily centered around the biosafety of micro-RNA drugs. We also addressed various technical difficulties associated with these drugs. Mr. Feng underscored the vital importance of the delivery system in this context. The fundamental features necessary for any effective delivery system include:
- Shielding the anti-miR/miRNA mimics from premature degradation once they enter the bloodstream.
- Positioning the anti-miR/miRNA mimics in close proximity to the target cells.
- Promoting cellular uptake of the anti-miR/miRNA mimics.
- Ensuring that the system does not trigger any immunogenic response.
- Incorporating materials that are both biocompatible and biodegradable.
Commercial aspects of micro-RNA drugs: The Vice President of Novozymes discussed the potential of micro-RNA drugs in the market and their operation. He urged us to focus on technology and R&D, and suggested collaboration with established pharmaceutical enterprises.
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Fig.7 visiting Novozymes Pharmaceuticals
Public Knowledge of New Biological Therapy (the safety of micro-RNA drugs)
We surveyed public attitude towards using new biological therapy on obesity and osteoarthritis, which means public attitude on the safety of micro-RNA drugs. By analyzing the survey result, we found that:
- Safety is the main concern that the respondents of the second survey have on the new therapy, with 80% mentioning it. We need pay more attention on ensuring and advocating the safety of our therapy.
- About 30% of our respondents in the second survey approve using synthetic biology methods to alleviate the impact of obesity on knee osteoarthritis, and 38% need more information before making a judgment. We hope to provide quality information to them and help them make an informed decision.
In conclusion, our analysis reveals that while a substantial portion of the public recognizes the connection between obesity and knee osteoarthritis, there is a clear need for enhanced education on the efficacy of weight management strategies, especially biological therapy. Addressing concerns related to safety and access to treatments is crucial for wider acceptance and utilization. Additionally, providing comprehensive information on synthetic biology methods can empower individuals to make well-informed decisions regarding their health. These insights will inform our educational initiatives, which can be read in our education page.
Implementation
Discuss Follow-up Experiments at CCiC
We took part in the 10th Conference of China iGEMer Community (CCiC). During the conference, we discussed our project concept, engineering design, and integrated human practices through a poster and online presentation. Experienced high school and Collegiate iGEM teams, familiar with miRNA, assisted us in transforming our follow-up experiments into protocol outlines. Notably, they guided us on how to identify the target gene of miRNA. We're thankful for the collaborative platform of CCiC that allowed us to interact with multiple teams!
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Fig.8 The booklet to introduce our project in CCiC
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Fig.9 The poster displayed in CCiC
Consulting Osteoarthritis and Obesity Experts on Implementation
During the second interview with Professor Guo, we gained a much deeper understanding on biological factors related to knee osteoarthritis. Professor Guo suggested doing an overlap between target gene that potentially have effects on osteoarthritis and gene that control knee cartilage to ensure the target gene that can cause knee osteoarthritis. If found, cultivate them in the cell test solution to verify. If it really has impacts on the cartilage, we need to first identify whether it inhibits or promotes the formation of cartilage. If it inhibits the formation, we can use an inhibitor to inhibit the RNA; on the contrary, if it promotes the formation, we can use a mimic to promote it. Both needs do cell experiments. In the future, direct intraarticular injection of miRNA can be tested by animal experiments and clinical trials.
Since the only method to cure osteoarthritis in obese subjects available now is to lose weight, the medicine has a much easier way, which means it will have a grand market. Despite this, the medicine still has some limits. First of all, inhibitor and mimic are synthetic drugs, which means they may have side effects. Second, micro-RNA is targeted at a specific gene, not a body part. So, it doesn't have a strong pertinence on knee and the cartilage. Which is where we need further research and improvement.
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Fig.10 Interviewing with Professor Zhao on the experiment implementation.
Please click the link to see the video interview in BiliBili:
Video: Interview with Professor Guo about osteoarthritis (2).
When queried about the process of confirming miR-3074's target genes in joint action, Professor Zhao explained the meticulous methodology. She described a multi-step approach involving miRNA profiling and cross-referencing with databases like TargetScan and Miranda. By identifying common target genes, such as SMAD4, associated with arthritis-related mechanisms like cartilage development, researchers can begin to uncover the gene interactions underlying joint health.
Regarding strategies to inhibit miR-3074, Professor Zhao outlined an experimental pathway. The verification process entails fluorescent labeling of extracellular vesicles and miRNAs to ascertain their joint entry capabilities. Utilizing dual luciferase assay with distinct fluorescence assists in verifying miR-3074's presence in joints and establishing specificity.
Professor Zhao emphasized the burgeoning potential of bioengineered therapies involving miRNAs. Drawing parallels with successful applications in countering drug addiction, she underscored the transformative potential of engineered miRNAs in treating conditions like osteoarthritis.
In terms of safety, ethical concerns and potential risks were discussed. While patient-derived extracellular vesicles seem promising due to their autologous nature, therapies using engineered miRNAs from external sources necessitate careful scrutiny for compatibility and adverse effects. Ongoing international research focuses on assessing the safety of these innovative approaches. In summary, the interview highlighted Professor Zhao Yue's pioneering work in deciphering miRNA targets and exploring their therapeutic utility in addressing obesity and osteoarthritis. The potential of bioengineered miRNA therapies offers hope for novel treatments, yet the safety and ethical dimensions remain integral to their future implementation.
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Fig.10 Interviewing with Professor Guo on the experiment implementation
Please click the link to see the video interview in BiliBili:
Video: Interview with Professor Zhao about ovese adipocyte exosomes (2).
Proposed Patients and Customers
Department of Health
The Department of Health's mission is advancing the health and well-being of all people by transforming the public health system through unique partnerships. They also take the lead in osteoarthritis treatment. When the department is informed of such a safe and efficient device, it will utilize such a product that can help its residents from the pain of OA.
Obese and OA Individuals
The primary target audience would be individuals who are already obese or overweight. These are at a higher risk of developing knee arthritis due to the excess stress on their joints. They may be seeking ways to manage their weight and reduce the impact on their knee joints. Obesity itself is an important risk factor for osteoarthritis disease, and weight loss can lead to improvements in other risk factors such as blood pressure and cholesterol levels. There's more to them than simply weight loss.
Business Plan
Publicity
Publicizing our project at iG 20
iG 20 was hosted by team LZU-CHINA, with participation from over 28 teams. We joined iG 20 to expose more people to our project. We produced four videos, which included an introduction to our school, an overview of our project, a promotion of our team, and a segment about synthetic biology. Through iG 20, we were able to attract more attention to our project and raise awareness about the impact of obesity on our joints.
Learn from Other High School iGEM Team on Integrated Human Practice
The high school online conference was hosted by BGI-mommath, Nanjing-Bioxstem, BS-United China, Squirrel, and the Nanjing-NFLS team. This conference focused on the integrated human practice, with the key emphasis being on "Integration". Each team provided an introduction to their project and their human practice, while iGEMer human practice instructor, Mr. Bao Yuhan, gave detailed instructions to enhance the effectiveness of our human practice efforts. The closing loop of “Background-----Public Attitude-----Implementation-----Publicity to make impact” is critical for every team and is also a benchmark goal of iGEM. It's essential to address the problems we encounter during our project and employ human practice activities to solve these issues. While education, inclusivity, and outreach are important tasks, they are not typically categorized under human practice work.
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Fig.12 High school meetup to exchange intergrated human practice
Learn from Similar Projects @ CRISPR team meetup
The CRISPR online conference was jointly hosted by Tongji-Software, WHU-China, AFMU, SMBU, CJUH-CLU-China, and the Nanjing-NFLS team. We were the only high school team to participate in this meeting. It was a fantastic opportunity for us to listen to and learn from the experiences of the other teams.
The CRISPR online conference was jointly hosted by Tongji-Software, WHU-China, AFMU, SMBU, CJUH-CLU-China, and the Nanjing-NFLS team. We were the only high school team to participate in this meeting. It was a fantastic opportunity for us to listen to and learn from the experiences of the other teams.
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Fig.13 Similar project team meetup to exchange
Partnership
Nanjing-China is a collegiate team aiming to use microbial cultivation technology to produce new dyed textile fibers as alternatives to traditional materials. Our shared residence in Nanjing brought us together, and our initial encounter at Nanjing University marked the inception of a productive collaboration. This partnership extended across various domains, encompassing the wet lab, modeling, and integrated human practices (IHP) facets of our respective projects. Throughout the year, our collaboration with Nanjing-China remained steadfast, driven by our common goal of optimizing the luciferase experiment, target gene searching modeling, western blot techniques, and cell transfection procedures that both of our teams were utilizing. Additionally, our joint efforts aimed to promote awareness about synthetic biology. In the realm of modeling, a significant exchange of knowledge occurred, as we shared insights into target gene searching modeling software with their dry lab team, while reciprocally gaining valuable insights from their modeling expertise (View detailed things in partnership page).
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Fig.14 The meetup with Nanjing-China team
In summary
Timeline of Our Integrated Human Practice
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The Last but Not Least
In fact, our student leader, Litong Wu, has personally benefited from this project. Through regular exercise and conscientious diet controls, he successfully reduced his BMI from 32 to 28. Beyond personal impact, we aim to utilize our project as a platform to amplify awareness about the implications of obesity and Osteoarthritis amongst our local community, and eventually, society at large.
Reference
1. Chakraborty C, Sharma AR, Sharma G, Lee SS. Therapeutic advances of miRNAs: A preclinical and clinical update. J Adv Res. 2020 Aug 29; 28: 127-138.
2. Chakraborty C, Sharma AR, Sharma G, Doss CGP, Lee SS. Therapeutic miRNA and siRNA: Moving from Bench to Clinic as Next Generation Medicine. Mol Ther Nucleic Acids. 2017 Sep 15; 8:132-143.
3. Moorkens E, Meuwissen N, Huys I, Declerck P, Vulto AG, Simoens S. The Market of Biopharmaceutical Medicines: A Snapshot of a Diverse Industrial Landscape. Front Pharmacol. 2017 Jun 8; 8:314.
4. Bajan S, Hutvagner G. RNA-Based Therapeutics: From Antisense Oligonucleotides to miRNAs. Cells. 2020 Jan 7;9(1):137.