Background
Amid the ongoing Coronavirus pandemic, people's health and lifestyles have been significantly impacted. Despite the full release of epidemic control measures in China at the end of 2022, many individuals are still being diagnosed with the virus and experiencing various degrees of symptoms without access to effective treatments. Due to various reasons, during this period of concentrated infections, hospitals have faced a shortage of medical resources, and the public has found themselves in a situation lacking effective treatment medications. There was even a surge in demand for fever-reducing medication like ibuprofen. Although the Omicron variant has a low mortality rate, and most individuals can recover within a week without any medication, the sense of helplessness during the infection period when there were virtually no available drugs remains fresh in people's memories. Therefore, we have decided to seek "new weapons" to combat the novel coronavirus to prepare for any potential future outbreaks.
In response, we have been gradually refining our research direction through literature review and expert interviews. We have initiated a study of the enzyme UGT72B2, which plays a crucial role in the production of antiviral compounds known as lignan glycosides found in Isatis Indigotica, a common Chinese traditional herb used in medications like the Lianhua Qingwen capsule. Lignan glycosides have shown the ability to inhibit virus replication and effectively treat both influenza and Coronavirus infections. On the other hand, we have conducted public surveys to gauge the public's attitudes toward our work. Based on their feedback, we have tailored our educational outreach efforts for future work.
Literature Review
In order to understand the current main methods for treating COVID-19, we have consulted the "Diagnosis and Treatment Protocol for Novel Coronavirus Infection (Trial Tenth Edition)" published by the Chinese government. This document addresses the pathogenic and epidemiological characteristics of the novel coronavirus (SARS-CoV-2), particularly the currently prevalent Omicron variant, as well as aspects of prevention, diagnosis, and treatment.
On one hand, the document reaffirms the importance of COVID-19 vaccination in reducing the risk of infection and disease, highlighting it as an effective measure to lower the occurrence of severe cases and fatalities. It recommends that all eligible individuals should receive vaccination, and those eligible for booster shots should do so in a timely manner.
On the other hand, the document provides detailed information on treatment methods for various scenarios, with a special mention of the use of traditional Chinese medicine.
Link to the document: https://www.gov.cn/zhengce/zhengceku/2023-01/06/5735343/files/5844ce04246b431dbd322d8ba10afb48.pdf
Conclusion: We plan to pursue our "new weapons" on two fronts: 1) Prevention through the improvement and development of vaccines, and 2) Treatment through the enhancement and research of pharmaceuticals.
Experts Interviews
To clarify the research direction of our project, we conducted interviews with six experts in fields related to COVID-19 treatment through various means, including phone calls, online video meetings, and in-person visits. These experts included specialists in virology, infectious disease physicians, respiratory specialists, traditional Chinese medicine practitioners, herbal medicine researchers, and pharmaceutical experts. We sought their opinions and advice to inform our project's focus.
Interview with Professor Binbin Ding, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology
As virus experts, we interviewed Professor Ding, hoping he could provide us with some guidance regarding our idea of developing a vaccine. Professor Ding went on to explain in detail the characteristics of the novel coronavirus, highlighting that, unlike DNA viruses, the novel coronavirus is an RNA virus with a high degree of variability, making it challenging to design effective vaccines for all subtypes. The replication mechanism of RNA viruses also adds to the complexity of vaccine development.
He mentioned that, despite the efforts of scientists and pharmaceutical companies worldwide, there are already various vaccines available for prevention of COVID-19. While their effectiveness might not be ideal, achieving a significant breakthrough in vaccine development is extremely difficult.
Professor Ding suggested that we should consider addressing the issue from a treatment perspective. For example, by preventing or inhibiting the release of the virus from infected cells, it becomes unable to replicate and spread within the human body after infection. As a result, it can be more effectively neutralized by the human immune system, leading to a reduction in disease severity and a faster recovery.
We were excited about this suggestion, and Professor Ding also recommended several relevant research papers to further explore this approach.
Interview with Dr. Huo, Infectious diseases Department, Peking University First Hospital
Dr. Huo, as an infectious disease specialist who has been on the front lines of the pandemic for the past three years, is believed to possess valuable clinical experience and insights that can offer valuable guidance for your new drug development efforts.
Dr. Huo candidly shared her experiences with you. Due to China's effective epidemic prevention and control measures over the past three years, she didn't have much direct exposure to COVID-19 patients for most of that time. However, during the recent surge in infections from late 2022 to early 2023, her team followed the "Diagnosis and Treatment Protocol for Novel Coronavirus Infection," while also using their clinical experience to assist patients.
She emphasized that in the treatment of COVID-19, both Western and traditional Chinese medicine have played positive roles. Western medications such as the combination of Nirmatrelvir/Ritonavir have shown good effects in the early stages of the disease but have significant side effects and cost issues. On the other hand, traditional Chinese medicine is relatively affordable and has shown good therapeutic effects. However, the clinical efficacy reports for traditional Chinese medicine are not as abundant and rigorous as those for Western medicine, which has limited its international recognition.
Dr. Huo sees synthetic biology as an emerging discipline that has made tremendous progress in recent decades. She acknowledged the importance of vaccines in controlling the spread of infectious diseases and the contributions of synthetic biology in improving vaccine immunogenicity, delivery, and safety. For example, gene editing technologies have been used to develop mRNA and DNA vaccines, significantly reducing the time required for vaccine development and production.
As a clinical practitioner, Dr. Huo expressed her hope that scientists involved in pathology research, drug development, and related fields will continue to provide increasingly effective tools to help healthcare professionals in their battle against diseases and improve patient outcomes. She firmly believes that emerging biotechnologies like synthetic biology will continue to drive advances in medical research, positively impacting the field of medicine.
Interview with Dr. Yang, Xiyuan Hospital, China Academy of Chinese Medical Sciences
Dr. Yang, with 20 years of experience in traditional Chinese medicine, possesses profound knowledge and insight into Chinese medicine and herbal remedies. We seek her advice from a traditional Chinese medicine perspective on our new drug development.
Dr. Yang believes that traditional Chinese medicine is a vast and profound field of study deeply rooted in Chinese culture, representing an important treasure trove of traditional Chinese culture. The effectiveness of Chinese herbal medicine is unquestionable. However, due to cultural differences, although there has been increasing Western acceptance and understanding of certain traditional Chinese medical practices like acupuncture and cupping in recent years, a complete acceptance and understanding of traditional Chinese medical theory remains challenging. Chinese herbal medicine is guided by traditional Chinese medical theory, and without understanding this theory, approaching the complex mechanisms of multi-component interactions in herbal medicine from a purely Western medical perspective can be difficult to grasp.
As a result, Dr. Yang suggests that if you wish to explore synthetic biology techniques in herbal medicine research, it might be more practical to start with the active components from a single herb, which is closer to the study of plant properties and is more likely to be accepted by the broader scientific community.
Interview with Professor Ying Xiao，Shanghai University of Traditional Chinese Medicine
Professor Xiao is an expert in the field of traditional Chinese medicine research, and she has provided us with many valuable insights regarding the development of drugs for treating COVID-19 using synthetic biology methods to produce effective components from Chinese herbal medicine.
Professor Xiao pointed out that the commonly used Isatis indigotica, also known as Indigo woad or "banlangen," actually contains antiviral active components. Academician Zhong Nanshan's team started researching this herbal medicine since the 2003 SARS epidemic and published several important papers in top-tier international journals, elucidating the mechanism of its virus-inhibiting activity. Apart from her own work, there have been many other teams conducting research on Indigo woad based on this foundation. She believes that our efforts in this direction should yield results, and she suggested that "protein engineering can be considered to modify the amino acid sites" to improve the enzyme and product performance.
The specific method involves protein engineering, which adjusts the spatial structure of the transferase by changing the amino acid sequence to enhance its catalytic effect and specificity. Another suggestion for delving deeper into this topic is to do some downstream work, such as protein engineering and the synthesis of rarer and more valuable downstream small molecule compounds. We can also continue to study and achieve the complete synthesis pathway of lignan glycosides in vitro so that E. coli can continuously produce the compounds we need.
Besides, Professor Xiao led us to the Chinese herbal medicine raw materials exhibition, which contained hundreds of dried herbs of different shapes and sizes. Next, we visited the herbal cultivation center, where plants used for further experiments are grown. It was a memorable experience where we learned extensively about the names of the herbs and the methods for their cultivation and harvesting and really broadens our horizons.
Interview with Prof. Zhengbing Lü, School of Life Sciences and Medicine, Zhejiang Sci-Tech University
Professor Lü is an expert in the pharmaceutical field, and we hope to get his advice on our idea to develop a new drug for treating COVID-19 based on effective antiviral components found in traditional Chinese medicine. If future drug development progresses, is it possible to achieve industrialization and push it into foreign markets with lower awareness of traditional Chinese medicine?
Professor Lü affirmed our idea and pointed out that currently, the pharmaceutical field consists of 70% chemical drugs and 30% biological drugs, with many new drugs being small-molecule drugs, which are difficult to develop and costly. Synthetic biology methods have already found many applications in the pharmaceutical field and offer unique advantages in the production of some high-value pharmaceutical components, with a very promising outlook. If our work has sufficient literature support, the feasibility is relatively high.
Of course, there is a long way to go from producing effective components to developing a new drug, which includes animal experiments and the three phases of clinical trials before obtaining drug approval and achieving actual market sales.
Although the components of the drug we envision are derived from traditional Chinese medicine, they are significantly different from traditional Chinese herbal medicine. In fact, in the Western world, Chinese herbal medicine is recognized as a natural product without any issues. Furthermore, we have well-founded theory and data supporting our effective antiviral components. As long as we can obtain drug approval, selling it in Western countries should not be a problem. With the development of the Chinese economy and the global spread of Chinese culture, including traditional Chinese medicine, our work could, in a way, contribute to the internationalization of traditional Chinese medicine and culture.
Interview with Dr. Xu，Shanghai Ninth People's Hospital
Doctor Xu is the head of the infectious disease department and specializes in the diagnosis and treatment of various infectious diseases. During the interview, we inquired about the necessity of continuing research on specialized drugs for COVID-19, sought Dr. Xu's opinions on our research project, and delved into topics of personal interest, such as the reasons behind the coronavirus's significant impact.
Dr. Xu's responses not only affirmed our project but also enriched our understanding of infectious diseases. She also offered valuable suggestions to enhance our performance in the final conference. She believed that it is still important to conduct research on antiviral drugs to cure COVID-19 and other types of coronaviruses because even though the pandemic is over, those infectious diseases still exist. Successful development of specialized drugs to treat coronaviruses could aid in future outbreak responses. She claims that our research project is very promising, but we should have a deeper understanding of how lignan glycosides inhibit viral replication and the mechanism by which UGTs catalyze the reaction. We should also take into consideration the long process of drug development when deciding on further steps. In addition, we learned more about why infectious diseases like COVID-19 are more common and more impactful. She explained that factors such as rainforest overexploitation, global connectivity, and high population density have accelerated the spread and mutation of infectious diseases. In addition, inadequate medical resources during the outbreak caused patients to wait a long time for medical treatment, which increased their psychological pressure.
Survey
Purpose
This survey aims to assess the public's experiences and perceptions regarding viral diseases like influenza and COVID-19. It covers aspects such as disease incidence, symptom severity, and coping strategies. Additionally, the survey seeks to gauge public sentiment regarding traditional Chinese medicine and our synthetic biology research project. Lastly, we aim to understand the public's interest in learning more about synthetic biology and their preferred medium for receiving information on this topic.
Analysis
In the first question, “How long has it been since you last had the flu or COVID-19?”, almost 90% out of the 405 respondents claim to have had the flu or COVID-19 within one year. Although the WHO declared an end to COVID-19 as a global health emergency, it is still quite common in China.
Figure 1.1 Q: How long has it been since you last had the flu or COVID-19? [Single Choice] A: Within half a year 38.02%; Half a year to a year 49.88%; One year to three years 7.41%; More than three years 4.69%
In addition, 80% of the respondents describe their degree of symptoms as moderate to serious, which shows that the flu and COVID-19 still have the prevalence and severity to pose a significant threat to public health. Common coping methods are self-medication (61%), resting without any medications (24%), and hospital consultation (13%). This suggests that a significant number of people rely on symptom-alleviation drugs and their immune systems to overcome these diseases. However, this will not work as well with high-age individuals who have deteriorating immune systems.
Figure 1.2 Q: How serious are your symptoms? [Single Choice] A: Very serious 5.68%; Serious 20%; Moderate 54.32%; Not serious 15.8%; No symptoms 4.2%
Figure 1.3 Q: What is your preferred way of dealing with the flu or COVID-19? [Single Choice] Green: Self-medication 61.23%; Yellow: Resting without any medications 24.44%; Blue: Hospital consultation 13.33%; Orange: Other 0.99%
Thus, approximately 80% of the respondents agree that it is necessary to continue to develop drugs that can effectively cure COVID-19, influenza, and other infectious diseases caused by viruses.
Figure 1.4 Q: Do you think there is a need to continue to develop drugs that can effectively treat infectious diseases caused by COVID-19, influenza, and other viruses? [Single Choice] Blue: Yes 79.75%; Yellow: Don't know/don’t care 11.11%; Green: No 9.14%
Despite choosing Western medicine more often during self-medication and prescriptions from doctors, 87% of the respondents expressed belief in traditional Chinese medicine and hope that it can be recognized internationally.
Figure 1.5 Q: Do you believe in traditional Chinese medicine and want it to be recognized internationally? [Single Choice] A: Believe, hope to be recognized 86.91%; Don’t know/don’t care 10.37%; Don’t believe, don’t want recognition 2.72%
The public’s choice on how to best increase international recognition was also what we were striving to do in our IGEM project, such as proving the scientific basis behind traditional Chinese medicine (53%) and increasing public awareness (31%).
Figure 1.6 Q: How do you think the international recognition of traditional Chinese medicine can be improved? [Single Choice] Green: Prove the scientific basis behind traditional Chinese medicine and publish the research results to reputable journals 52.84%; Blue: Increase public awareness 31.25%; Yellow: Combination of traditional Chinese and western medicine 1.14%; Light green: Other 0.57%
After briefly introducing our project in the survey, about 90% of the respondents believed that our project was meaningful and would support our work. Those who were neutral in their views accounted for roughly 9% of the respondents, with approximately 1% holding other views.
Figure 1.7 Q: Do you think our project is valuable and do you support our work? [Single Choice] Blue: Valuable, support 90.12%; Yellow: I don’t know/It doesn’t matter 8.89%; Green: Not valuable, do not support 0.99%
Although only about half of the respondents had heard about synthetic biology, about 73% of them hoped to learn more about this subject. The data shows that most of the public is looking forward to the future development of synthetic biology and is eager to discover more in this field.
Figure 1.8 Q: Have you ever heard of synthetic biology? [Single Choice] Blue: Yes 52.1%; Green: No 47.9%
Figure 1.9 Q: Would you like to learn more about synthetic biology? [Single Choice] A: I would like to 72.84%; It doesn’t matter 23.7%; I would not like to 3.46%
Of those who hoped to learn more, the top three mediums they wanted to learn from were watching scientific videos (72%), viewing WeChat official account tweets (55%), and reading relevant science research articles (33%). When choosing how to educate the public about synthetic biology, we can choose a more targeted medium that is more appealing to them.
Figure 1.10 Q: How would you like to learn more? [Multiple Choice] Green: Watch scientific videos 72.2%; Yellow: View WeChat official account tweets 55.25%; Orange: Read relevant science research articles 33.22%; Light green: Participate in scientific activities 31.53%; Gray: Receive scientific pamphlets 25.42%; Blue: Read leaflets and posters 20.68%
Summary
Overall, viral diseases like influenza and COVID-19 continue to have a significant impact on the public, highlighting the need for effective treatments.
The survey results align with our previous outreach efforts and help us choose the right medium for future public education initiatives.
Notably, 90% of respondents expressed a desire to globalize traditional Chinese medicine and showed support for our project, reinforcing its significance to the public.
Integrated Human Practices
Fig.1 The Relationship between HP research and clarity process of the project direction
We have conducted the research work of HP carefully using three methods: literature review, expert interviews, and public surveys. By reading important documents, including those from the Chinese government, collecting advice from expert interviews, and analyzing data from public surveys, the results show that the government, experts, stakeholders, and the general public all believe that our project - using synthetic biology methods to produce enzymes for catalyzing the production of antiviral components in the traditional Chinese herb "Isatis indigotica" - has practical significance and promising prospects. It not only serves as a powerful "weapon" for humanity to cope with potential future RNA virus outbreaks but also has the potential to contribute to the global recognition of traditional Chinese medicine.
Throughout the HP research process, the results from literature reviews and expert interviews have continuously helped us clarify the project's research direction. The results from public surveys have not only strengthened our confidence in the project but also guided us toward the direction of public education and outreach activities, laying a solid foundation for the successful implementation of these activities (See Fig.1).
(1) Clarifying the Research Topic
After brainstorming, we initially formed the project's primary direction (Idea 1.0): to find or create a "new weapon" to combat COVID-19 or similar RNA viruses, to help ourselves, our families, friends, and all of humanity to deal with potential future outbreaks. To find this new "weapon," we downloaded the official "Diagnosis and Treatment Protocol for Novel Coronavirus Infection"[1] from the Chinese government website (See Fig.2). By reading this crucial document, we realized that the two most important aspects of combating COVID-19 are prevention and treatment. Prevention mainly includes two approaches: proactive protection through vaccination and passive protection through wearing masks, frequent handwashing, social distancing, and more. Treatment provides detailed plans for different conditions, including both Western medicine and traditional Chinese medicine. Based on this, we refined the initial project proposal (Idea 1.0) into two directions: improvement and development of vaccines for prevention (Idea 1.1) and improvement or development of drugs for treatment (Idea 1.2).
Fig.2 Screenshot of Chinese government website
To support the argument for the direction of vaccine improvement and new vaccine development (Idea 1.1), we interviewed Professor Ding, a virology expert. Professor Ding explained the challenges of RNA virus vaccine development and advised against delving deeper into this direction. Instead, he recommended considering starting from the virus itself to explore new treatment directions (Idea 2.0) and provided several references[2-4] for our consideration. We accepted Professor Ding's advice and terminated the exploration of the vaccine improvement and new vaccine development direction (Idea 1.1). After discussions and assessments with our teachers, although it was a difficult decision, we also abandoned the new treatment direction starting from the virus itself (Idea 2.0) due to the objective limitation.
To substantiate the direction of drug improvement or new drug development (Idea 1.2), we interviewed two doctors, infectious disease physician Dr. Huo and traditional Chinese medicine practitioner Dr. Yang. These experienced doctors shared their frontline experience in diagnosing and treating COVID-19 and discussed their perspectives on the combination of Western and traditional Chinese medicine in treating COVID-19. Based on their advice, we focused the drug improvement or new drug development direction (Idea 1.2) in the field of traditional Chinese medicine (Idea 3.0).
Therefore, we conducted interviews with traditional Chinese medicine expert Professor Xiao and pharmaceutical expert Professor Lü to seek their advice. Both professors provided us with significant encouragement and assistance. Professor Xiao suggested that we focus on the relatively well-established "Isatis indigotica" and provided a batch of important research papers[5-9], including papers from the team of Academician Zhong Nanshan, for our reference. While Professor Lü informed us about the long journey from drug development to market approval, he dispelled our doubts about traditional Chinese medicine's global recognition, leaving us greatly encouraged. This ultimately led to the formation of our project (Idea 4.0) - using synthetic biology methods to produce enzyme UGT72B2 to catalyze the production of antiviral compounds, lignan glycosides.
(2) Wet lab work and education plan design
Professor Xiao's advice and assistance played a crucial role in the design of our experimental plan. Subsequently, Dr. Xu's suggestions, such as the need for a deeper understanding of how lignan glycosides inhibit virus replication and the mechanism of UGT-catalyzed reactions, were very helpful in our reflection and improvement of the work plan.
Our public survey not only greatly encouraged us to advance the project but also provided us with a relatively in-depth understanding of the public's attitudes toward synthetic biology, the novel coronavirus, and traditional Chinese medicine. This, in turn, helped us formulate reasonable education and outreach plans, resulting in a positive impact.
Reference:
[1] "Diagnosis and Treatment Protocol for Novel Coronavirus Infection (Trial Tenth Edition)".https://www.gov.cn/zhengce/zhengceku/2023-01/06/content_5735343.htm
[2] Yuan Z., Hu B., Xiao H., Tan X., Li Y., Tang K., Zhang Y., Cai K., and Ding B. (2022). The E3 Ubiquitin Ligase RNF5 Facilitates SARS-CoV-2 Membrane Protein-Mediated Virion Release. mBio 13, e03168–03121. doi:10.1128/mbio.03168-21.
[3] V’kovski P. et al. (2021). Coronavirus biology and replication: implications for SARS-CoV-2, Nature Reviews Microbiology, 19: 155–170.
[4] He M-L, et al. Kinetics and synergistic effects of siRNAs targeting structural and replicase genes of SARS-associated coronavirus. Febs. Lett. 2006;580:2414–2420. Doi
[5] Li, Jing, Zhou, Beixian, Li, Chufang, et al. Lariciresinol-4-O-β-D-glucopyranoside From the Root of Isatis indigotica Inhibits Influenza A virus-induced Pro-Inflammatory Response[J]. Journal of Ethnopharmacology, 2015, 174(Complete): S037887411530101X.
[6] Beixian Zhou, Jing Li, Xiaoli Liang, et al. Transcriptome profiling of influenza A virus-infected lung epithelial (A549) cells with lariciresinol-4-O-D-glucopyranoside treatment[J]. Plos One, 2017, 12(3): e0173058.
[7] Zhong, Nanshan. Antiviral activity of Isatis indigotica root-derived clemastanin B against human and avian influenza A and B viruses in vitro[J]. International Journal of Molecular Medicine, 2013, 31(4).
[8] Jianmin Chen, Jasdeep K. Saggar, Paul Corey, et al. Flaxseed and Pure Secoisolariciresinol Diglucoside, but Not Flaxseed Hull, Reduce Human Breast Tumor Growth (MCF-7) in Athymic Mice[J]. Journal of Nutrition, 2009, 139(11):2061-2066.
[9]Yang Z, Wang Y, Zheng Z, Zhao S, Zhao J, Lin Q, Li C, Zhu Q, Zhong N. Antiviral activity of Isatis indigotica root-derived clemastanin B against human and avian influenza A and B viruses in vitro. International Journal of Molecular Medicine, 2013, 31(4): 867.