Integrated Human Practices

1. Introduction

1.1 Overview

There are many microorganisms around us, in the air, in the water, and on every object we touch. A large part of them are bacteria, and a large part of them can cause us to get sick. The most effective treatment for infectious diseases in humans is still the use of antibiotics. Antibiotic drugs have saved hundreds of millions of lives in the past hundred years, but there are also some problems because of the single treatment method.

1.2 Antibiotic

Since the discovery of penicillin by British doctor Fleming in 1929, more than 100 kinds of antibiotics have been developed in more than 12 classes.

Between 1940 and 1962, the number of antibiotics increased by leaps and bounds, with more than 20 classes of antibiotics entering the market and gaining clinical application [2]. But in the decades since, only two classes of antibiotics have been developed and used [3-5]. This means that bacteria have been fighting these antibiotics for nearly a hundred years, and we need these drugs to continue to work for another hundred years.

1.3 Antibiotic Resistance

Over a century of antibiotic use and abuse, bacterial resistance to antibiotics is also on the rise. In recent years, bacteria have emerged that are resistant to multiple antibiotics, and even "superbugs" that are resistant to all current antibiotics. According to the data from China Antimicrobial Resistance Surveillance System (CARSS), the resistance rate of Pseudomonas aeruginosa to carbapenems, such as imipenem, was 17.7% in China in 2021. The resistance of Acinetobacter baumannii to carbapenems was as high as 54.3%, and there was an increasing trend year by year [6]. Drug-resistant bacteria pose an extremely difficult problem for patients and doctors alike. According to the AMR Action Fund, more than 1.2 million people die each year from diseases linked to bacterial resistance [7]. In 2017, the World Health Organization listed carbapenem-resistant P. aeruginosa as Priority 1 on its list of priority pathogens for new antibiotic development [8]. It is urgent for us to develop new antimicrobial drugs.

2. Our solutions

2.1 Reasonable use of antibiotics

Why: The use of antibiotics is the primary driving force leading to antibiotic resistance in bacteria, and bacteria will only accelerate the speed of resistance selection in the environment of a large number of antibiotics, which is crucial for the rational use of antibiotics.

How: Make reasonable diagnosis and treatment plans for different patients, reduce the use of broad-spectrum antibiotics as much as possible, and use antibiotics reasonably and restrictively.

2.2 Development of new antibacterial drugs

Why: The long-term use of existing antibiotic drugs has caused bacteria to produce a corresponding rich resistance mechanism, we need some new mechanisms of antibacterial drugs, to effectively kill bacteria.

How: Increase investment in antibacterial drug research and development, adopt new mechanisms to develop corresponding antibacterial drugs, and promote the transformation and utilization of scientific research.

2.3 Controlling Hospital Infections

Why: P. aeruginosa is an opportunistic pathogen that only infects us when our immunity is low or when our barrier system is faulty. We are most likely to be infected with P. aeruginosa during medical treatment.

How: Strengthen the control of the microbial environment in the hospital, publicize reasonable hospital sense knowledge to patients, and improve their awareness of protection.

2.4 Control of primary disease

Why: P. aeruginosa infection is often caused by the combination of primary disease, and the control of primary disease is conducive to the prevention and treatment of P. aeruginosa .

How: Enhance the treatment of the primary disease and control the wound.

2.5 Maintain a healthy lifestyle

Why: The immune system is fundamental to our ability to kill bacteria and other microbes, and a healthy lifestyle helps strengthen the immune system.

How: Promote an active and healthy lifestyle by promoting a balanced diet, adequate sleep, and moderate exercise.

3. From upstream to downstream

3.1 Understanding needs: clinical investigation

In order to understand antibiotic resistance, our team especially came to the hospital for research. We contacted doctors in the respiratory department, infection department, and acute and critical care department of Southern University of Science and Technology Hospital and Shenzhen First People's Hospital to get into the clinical practice to understand their needs.

3.1.1 Infection Department Dr. Xiao Yumei

Dr. Xiao first introduced to us the clinical infection status of P. aeruginosa : According to the national Bacterial Resistance Monitoring report in 2021, the bacterial isolation rate of P. aeruginosa in the hospitals included in the data accounted for the fourth place of all isolates, second only to Staphylococcus aureus , and has become a more difficult class of drug-resistant bacteria in clinical treatment. So she praised our project and encouraged us to work on it. At the same time, Dr. Xiao also emphasized that clinical data collected by several hospitals also showed similarities with this data, confirming that P. aeruginosa is a common cause of infection. The drug resistance of P. aeruginosa is also worthy of attention. Although the data in recent years show that the drug resistance to carbapenems such as imipenem and meropenem is declining year by year, it still brings no small trouble to treatment. However, the resistance rate of P. aeruginosa to polymyxins is generally low, which makes polymyxins an important drug in the treatment of its infection.

As an infection doctor, Dr. Xiao believes that the current problem of bacterial antibiotic resistance is still more serious, and sometimes it is difficult for doctors to make choices, and may even seriously affect the treatment effect. For the treatment of such bacterial infections, drug susceptibility tests are generally carried out, and single or combined drug therapy is carried out based on clinical characteristics and drug susceptibility test results and supplemented by other means (such as atomization, etc.) to achieve therapeutic effect and minimize damage.

3.1.2 Dr. Luo Xiangying, Department of Emergency and Critical Care Medicine

Professor Luo first summarized the current clinical use of phages in Xili Hospital: At present, the hospital has carried out phage therapy for some patients in the ICU ward, and the patients are generally over 70 years old and have strong resistance to pathogenic bacteria in the body. In the course of treatment, phage therapy against Acinetobacter baumannii was found to be more effective, while phage therapy against P. aeruginosa and Klebsiella pneumoniae was more limited. In the 5 cases that have been treated, a total of three cases have been successfully treated and discharged with phage therapy, and the prognosis is good without recurrence, it can be said that the success rate of this therapy is quite high.

Next, Teacher Luo introduced the principle of this treatment method in detail to us: the hospital took the patient's sputum as a sample, extracted and cultured P. aeruginosa from it, and then the hospital sent the strain to the Chinese Academy of Sciences, which screened the strain, searched for the corresponding phage for the strain, and took atomization treatment twice to three times a day after obtaining the phage.

Of course, Teacher Luo also pointed out the shortcomings of this treatment method, and hopes that it can be improved in the future: phage therapy is currently used by the main body of the elderly, due to the decline of resistance of the elderly, the rejection of foreign objects such as phages in the body is less, can have a better effect; In young people, immune rejection may cause symptoms such as fever, which makes phage therapy less effective. In addition, the current therapy is relatively limited. According to the analysis of current successful treatment cases, the treatment of Acinetobacter baumannii is the most effective, while the effect of other strains is not ideal. Moreover, the corresponding phage design is specific and cannot be widely used in a large number of strains.

3.2 Determination of ideas: theoretical feasibility

3.2.1 Professor Yang Liang

Professor Yang Liang is a professor in the School of Medicine of Southern University of Science and Technology, and his direction happens to be about P. aeruginosa , so our team found Professor Yang Liang and asked him about his views and suggestions on P. aeruginosa .

He first said yes to the idea that we want to target the currently more difficult hospital bacteria – P. aeruginosa . Then Professor Yang Liang began to introduce some properties of P. aeruginosa to us. He says one of the most important mechanisms of resistance to P. aeruginosa relies on the production of biofilms that attach to the body, preventing the antibiotics we use and the body's immune system from killing effectively. One of the most important pathways of biofilm production is the regulation of C-di-GMP signaling molecules. If we can target this molecule and control its ability to regulate biofilm production, we can effectively inhibit its tolerance to immune destruction and reduce its antibiotic resistance.

Professor Yang Liang then mentioned that pf4 phage is a class of phage that can specifically target P. aeruginosa . It is a mild phage that does not directly lyse the P. aeruginosa to release endotoxin, and P. aeruginosa has the homologous gene of pf4. By using this principle, we can carry out homologous transformation and thus carry out gene editing.

From Professor Yang Liang, we determined our experimental idea, that is, using the engineered pf4 phage to specifically target P. aeruginosa and inhibit the production of its biofilm.

3.2.2 Professor R.E.W. (Bob) HANCOCK

R.E.W. (Bob) HANCOCK is a Professor in the Department of Microbiology and Immunology at the University of British Columbia, Director of the Centre for Research in Microbial Diseases and Immunity (CMDR), and Canada Research Chair in Health and Genomics. Our team reached out to Professor R.E.W. (Bob) HANCOCK for guidance on our project.

When we introduced our project to Professor Hancock, he agreed with us and encouraged us to go ahead with it, praising our project as advanced. Since during the investigation, our team found that phages are foreign bodies that may cause immune system responses, we asked Professor Hancock for his opinion. Bob suggests that we consider in vivo trials, but cautions that such trials require a long period of training to be approved. He shared research from his lab: They first isolated P. aeruginosa and screened these bacteria for the effects of antimicrobial peptides in biofilm growth. Since the pathological change triggered by the biofilm is inflammation, they also tested for anti-inflammatory activity. As a second step of screening, they developed organoid models of artificial skin burns and sinusitis infections that allow biofilm growth and tracking, and monitor infection non-invasively, as well as inflammation by detecting the production of pro-inflammatory cytokines in these animal models.

In the process of research, we also encountered a series of difficulties. First of all, we encountered difficulties in the results of Western Blot experiments. Although we designed the target gene fragment with the flag tag, it did not match the expected results. Bob suspects that this may have something to do with the fact that the tag we use, the flag tag, is not an ideal way to tag, even though it is widely used. In addition, since no specific bands appear in the Western Blot, this means that the corresponding protein expression may not be obtained. To this end, Bob suggested that we first detect the mRNA by RT-PCR to determine whether the target sequence has been successfully transcribed. If successfully transcribed mRNA is obtained, then the problem of protein expression is solved. It is worth noting that phages have been natural enemies of bacteria for thousands of years, but bacteria have also evolved mechanisms to resist phages. In order to verify this idea, we first need to correctly induce pf4 expression in phage, and induction with arabinose is feasible. Next, the tag should be inserted at the appropriate location and verified by PCR that the target sequence has been inserted at the correct location. After isolating the phage and confirming whether there is an inserted sequence in it, we finally need to use RT-PCR to ensure that the target sequence has been correctly transcribed. The unsatisfactory results of our western blot experiment may be due to the fact that the repressor protein on pf4 binds to the mRNA transcribed by pf4. He suggested searching for data to destroy this repressor protein to achieve protein expression.

Regarding phage therapy as a possible cause of microbiome disruption, Bob believes that phage therapy is safer than antibiotics alone. Phage therapy is considered to be more selective than antibiotics. Because phages are typically host-specific, attacking only specific species or strains of bacteria, this means they are less likely to disrupt microbial communities broadly, unlike some broad-spectrum antibiotics, which may disrupt the normal gut microbiota, resulting in an imbalance. In addition, the advantages of phages are their large size and low mobility in the body, making them only targeted against local infections, which may also be a disadvantage compared to antibiotics that can treat systemic infections. Therefore, specialized delivery methods, such as the use of air solvents in pulmonary cystic fibrosis, have become particularly important.

Bob advised us on the design and method of the experiment. He points out that often using only a single phage may be only moderately effective, so he recommends using a mixture of multiple phages, known as "cocktail therapy." This approach has been widely used because, just as bacteria can develop resistance to antibiotics, they can also alter surface receptors to resist phage invasion.

3.2.3 Professor Mark Sutton

Professor J. Mark Sutton is the subject lead at UKHSA Porton Down and Professor of Antimicrobial Therapy at the Institute of Pharmaceutical Sciences, King's College London. His research interests include pathogenic mechanisms and virulence of bacteria, the development and evaluation of new antibacterial drugs, rapid detection and diagnosis systems, etc. His team uses a variety of technologies to develop solutions to real-world problems, working closely with end users, the academic community, and the business community. The group takes an interdisciplinary approach to working with various groups in the life sciences, chemistry, and physical sciences. He is the author of more than 100 peer-reviewed publications and an inventor of 19 international patents. Our team went to the UK to visit Professor Mark Sutton, hoping to learn from his experience.

We presented our experimental design ideas to our professor, who patiently listened to our presentation and spoke highly of our project. He said that phage therapy is currently a project of great interest to the academic community and has also attracted wide attention in community medicine, and encouraged us to continue to do it.

Next, the professor raised his questions about many points of interest in our project, and we gave our own answers.

Mark: Why did you choose the phage pf4 for your study?

SUSTech-MED: First, because pf4 is a mild bacteriophage, it does not break down the bacteria as virulent bacteriophages do, causing bacterial fragments to break up everywhere. It can survive in the bacteria for a long time in the form of former phages, and carry its genes to the outside; Secondly, P. aeruginosa itself carries the pf4 gene, which makes it easier for us to modify P. aeruginosa .

SUSTech-MED: We would like to ask what you think about how to verify the connection between the different forms. We detected the relevant expression at the gene level, but there was no relevant result in the Western blot experiment.

Mark: That's a difficult question to answer. In fact, the relationship between pf4 and P. aeruginosa is relatively complicated. If you carry out genetic modification, it may have some unknown effects on P. aeruginosa , which may make the analysis of your experimental results very complicated. The reason for this is more complicated, but I would suggest that you remove the primary antibody first, use only the secondary antibody to see what you get, and then gradually remove the secondary antibody to see what you get.

SUSTech-MED: The literature suggests that mild phages cause inflammation, but we were unable to confirm this through in vitro experiments. Would you recommend in vivo testing?

Mark: This is a controversial area and not everyone agrees on this; Do you have a specific paper on pf4? You should observe the migration of neutrophils in vitro. That might answer your question.

SUSTech-MED: Are there any mature experimental models in this area that can be directly applied to our projects?

Mark: Not sure what the specific requirements are here. We have built biofilm models and in vivo infection tests that you can learn about and then choose the one that suits you.

3.2.4 Preparation for mathematical modeling

During the summer vacation, the HP group also had targeted learning. In our team, few students are responsible for modeling and mathematical analysis, and it is very difficult for us to build a model from scratch or even to establish a model for a strange experiment without any experience in competition. So, we actively seek experience and guidance from various teams and seniors. We contacted Tongkai Zhang and asked him about ordinary differential equations and flux balance analysis, and he gave us very detailed answers.

ODE (Ordinary Differential Equation):

In the model, multiple graphs were generated based on different initial values. Instead of creating multiple similar graphs, consider using 3D graphs or force field diagrams to represent different conditions. These types of graphics can better illustrate continuous changes in the system's state. It provides a more comprehensive explanation of the experiment's purpose, including the significance of calculating the target lysis volume and infection rate for bacteriophages. The bivariate curve graphs are somewhat ordinary and may benefit from more visually appealing options, such as 3D graphs and force field diagrams.

FBA (Flux Balance Analysis):

Rather than presenting the original results using numerical lists, consider plotting the data as curves for a more intuitive representation. But please be cautious about imposing constraints on the upper and lower bounds of flow; these constraints might be overly stringent and should be used judiciously.

3.3 Production possibility: Survey of pharmaceutical enterprises

In order to have a better understanding of the clinical transformation of the research results of the project, our team flew to Shanghai to conduct research in pharmaceutical enterprises. We chose Hepu Pharmaceutical as our visit destination. As the first batch of key entrepreneurial enterprises in the Pudong New Area of Shanghai, it focuses on the research and development of original new drugs and has the overall research and development capability from new drug concept design, preclinical research, and pilot study to clinical research and mass production of new drugs.

We were warmly received by General Manager Han of Hep Pharmaceutical, who first took us to visit the science and technology park. Before the official visit, we were briefed on the safety and hygiene regulations and the protective measures that must be followed, including the wearing of specific protective clothing, safety glasses, masks, etc. First, we were taken to the warehouse where raw materials were stored and examined, where we saw mice raised with different genotypes. Afterward, the staff guided us into the production area of the company to watch the different stages of drug production, explaining the relevant technical principles and specific functions of the instrument in detail. In the production area, we also saw the quality control laboratory, which is the place to ensure that the product meets the quality standards, where the laboratory technicians demonstrated how to conduct sample testing and analysis. After the production is completed, we go to the packaging area to see how the drug is packaged into the final product, including the drug filling, labeling, and packaging process. Of course, in order to ensure the hygiene and sterility of the pharmaceutical environment, we also learned about the cleaning and disinfection process of the production area.

As the tour of the pharmaceutical process came to an end, we seized the opportunity to interact with professionals, learn about the pharmaceutical companies' research and development programs and their marketing strategies, and ask questions and exchange views with President Han and representatives of the technical staff.

During the exchange, Mr. Han gave valuable suggestions after hearing the introduction of our project and the purpose of this visit.

Starting from the five stages of drug research and development, Mr. Han talked with the two students about the possible confusion and factors to be considered in the transformation of results. He said that the transformation of results is different from scientific research, which focuses on cutting-edge innovation, while the former pays more attention to the effectiveness of clinical treatment and the safety of the human body, and needs to strictly control the toxicity of each component and each dose into the body. With regard to bacteriophage therapy studied by the SUSTech Med team, he pointed out that the evaluation of MIC (minimum inhibitory concentration) and MDC (minimum bactericidal concentration) on the one hand, and the identification of phage titers and activity in clinical reagents on the other hand, are extremely important for antimicrobial drug development. Our team built a mild bacteriophage specific to P. aeruginosa . At present, people usually use a 'cocktail' preparation, that is, a combination of multiple bacteriophages to play a role in bacteriostasis or bactericidal effect. If the phage can be combined with other bacteriophages in the transformation of results, it can also be regarded as a broadening and optimization of clinical application. In addition, the preservation conditions of purified bacteriophages, the effects of repeated freezing and thawing on their activity, and appropriate drug administration should also be considered.

Of course, General Han also affirmed that phage therapy is a new means of choice to fight bacterial infections, although it is difficult to replace antibiotics as first-line drugs, it can reduce its use on the basis of antibiotics, shorten the use cycle, and show great application prospects in this increasingly serious drug resistance "post-antibiotic era", which also makes our team confident. Continue to explore deeper into the subject!

Finally, after sufficient research, we summarized the influencing factors of all stakeholders as follows:

4. References

1. https://www.compoundchem.com/2014/09/08/antibiotics/

2. Powers J. H. (2004). Antimicrobial drug development--the past, the present, and the future. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases, 10 Suppl 4, 23–31.

3. Butler, M. S., & Buss, A. D. (2006). Natural products--the future scaffolds for novel antibiotics?. Biochemical pharmacology, 71(7), 919–929.

4. Hair, P. I., & Keam, S. J. (2007). Daptomycin: a review of its use in the management of complicated skin and soft-tissue infections and Staphylococcus aureus bacteraemia. Drugs, 67(10), 1483–1512.

5. Zappia, G., Menendez, P., Monache, G. D., Misiti, D., Nevola, L., & Botta, B. (2007). The contribution of oxazolidinone frame to the biological activity of pharmaceutical drugs and natural products. Mini reviews in medicinal chemistry, 7(4), 389–409.

6. http://www.carss.cn/Report/Details?aId=862

7. https://www.amractionfund.com/about

8. https://www.who.int/zh/news/item/27-02-2017-who-publishes-list-of-bacteria-for-which-new-antibiotics-are-urgently-needed

Education and Communication

Knowing that the exchange of education and knowledge around the project is valued by the iGEM judging criteria is a truly meaningful thing for our team. Nosocomial infection is a serious challenge facing the world at present, the background of its aggravation is closely related to multiple factors. The rise of drug-resistant bacteria, increased global mobility, aging populations, modern medical complexity, and equipment use all make infection control in healthcare facilities more difficult. The infection of P. aeruginosa , one of the most common bacteria in hospital infections, can be extremely resistant to conventional antibiotics and make it difficult to treat. Patients are often susceptible to these bacteria when their immune systems are compromised or after surgery. Such infections not only increase the burden of hospital infections but also make the patient recovery process more complicated and difficult. Therefore, the control and prevention of P. aeruginosa infection is essential to improve patient safety and reduce hospital infection rates. So, we urgently need to raise awareness, not just of the medical staff and patients in the hospital, but of the hundreds of thousands of people living in the community.

In addition to raising awareness of these issues, we hope to show how biology, and more specifically synthetic biology, can be applied to solve these problems, and that our work using mild phage therapy will continue to come into the public eye and be recognized by more hospital staff. By doing so, we seize the opportunity to promote this area of research as well as the iGEM competition.

In the past year, we have achieved results in various areas of education and communication. We not only promote our projects through push notifications and videos, but also actively collaborate with other iGEM teams, interview professionals in the pharmaceutical industry, and contribute to primary and secondary education. Our interactions with universities have also strengthened our presence in academia. We will continue our efforts to communicate scientific knowledge to a wider audience and contribute to a better future.

1. For diverse propaganda & education

1.1 Posts and video

First of all, we actively popularize the orientation content of our project for the public to help more people understand the scientific knowledge about hospital sense, P. aeruginosa , and phage. Through the above three aspects, we introduce our mild phage treatment method and the treatment mechanism for P. aeruginosa in the push of the introduction of the team project. To expose more communities to this new class of treatments for resistant microbes.

Of course, in addition to the push of our public account, we also produced and published our popular science videos on Bilibili, the largest video platform in China, including the introduction of synthetic biology, the attention and prevention of hospital sense, antibiotics and bacterial resistance, and phage therapy. Our purpose is not only to let medical staff and patients know about new therapies. It is also necessary to let more people understand and pay attention to the prevention of microbial infections.

In order to further expand the propaganda influence in China, we have taken a series of local propaganda measures. We have created dedicated official accounts and pages on major social media platforms, such as WeChat public account, YouTube, TikTok, etc. These platforms provide us with a wider audience and convey scientific knowledge through different forms of content (such as short videos, graphic push, live broadcasts, etc.), attracting more people's attention.

When we promote in the video and push categories, we adopt a number of strategies and approaches to ensure that our popular science content attracts the interest of a wider audience:

(1) Narrative Content: We are committed to bringing scientific knowledge to life and storytelling. We make it easier for viewers to relate to the content by telling real-life cases of hospital care, patient discharge experiences with phage treatment, and insights from medical experts.

(2) Images and animations: In order to better explain complex concepts, we use images and animations. This helps with visual learning, making it easier for viewers to understand how phage therapy works and the mechanisms of P. aeruginosa .

(3) Interactivity: We encourage the audience to participate in the interaction, such as leaving comments after the video push, asking questions, interspersing interactive plug-ins in each video, etc. This interactivity helps build closer communities, enabling our viewers to feel their voices are heard and to discuss relevant topics in depth.

(4) Regular updates: We regularly release new popular science content to keep the interest of the audience and update the progress of our experimental content and communication activities simultaneously. The continuous provision of new information helps to ensure that our audiences are kept up to date with the latest developments and research findings.

(5) Pay attention to feedback: We actively pay attention to audience feedback and suggestions to continuously improve our content and promotional strategy. This helps us better meet the needs and expectations of our audience.

Through these videos and push strategies, we strive to ensure that popular science knowledge is communicated in an engaging way so that more people understand the importance of phage therapy and P. aeruginosa infection, as well as how to prevent the problems of illness and bacterial resistance. Our goal is to provide accessible and interesting science education to the general public, promoting public interest in microbiology, synthetic biology, and medicine.

1.2 Game!

In order to let more people know about synthetic biology and our project, we have been trying to innovate in different ways, publicizing it in a form that everyone can accept, so that more people can be influenced and even join us. This time, our innovative way of using games can attract more people's attention and lower the threshold of science popularization.

We use the life of bacteria as the underlying logic of the game, giving the most important parameters of bacterial life: vitality, virulence, infectivity, and drug resistance. Let the game participants freely choose their parameters to obtain different bacterial life trajectories. At the same time, we set up many random events in the game, such as infected people washing their hands, and different parameters corresponding to different results, so as to increase the fun and complexity of the game. There is scientific knowledge we want to convey under different events, such as the important means to prevent P. aeruginosa is to maintain personal hygiene at all times and wear a good mask and timely protection.

Once the game was launched, it gained a lot of people's love and was widely spread around us, which made it easier for more people to get our science content and let more people understand synthetic biology.

1.3 Education brochure

In order to better promote our project and the philosophy behind it to everyone, we have created an educational brochure. Whenever we hold an event, we will give this booklet as a gift to the audience to achieve the purpose of publicity. Our brochure contains an introduction to the iGEM competition, our team, PI, topics, and application progress. Many friends like this brochure.

2. Communication and cooperation with other teams

Our team hopes to inspire and learn from each other in our communication. So we talked extensively with other teams

We have carried out many online and offline communication activities with other iGEM teams. Through many exchanges, on the one hand, we have introduced our project to expand its influence, and on the other hand, we have learned valuable lessons and experiences from other teams, so as to achieve the effect of promoting mutual learning, learning from each other's strengths and weaknesses, and making common progress. The following is the exchange meeting we participated in and the exchange experience with other teams:

2.1 The 7th South China Regional Meeting

On May 21st, we went to Shenda Lihu Campus to participate in the South China Exchange Meeting hosted by the iGEM team of Shenzhen University and had a friendly exchange with the iGEM team from sister schools in South China. Among the dozen teams, there are not only teams from the same university as us but also several teams from other regions. In addition, there are high school and foreign language school teams. After the official opening of the meeting, we came to the eight-minute session where each team introduced their project in detail. Listening to other teams, we broadened our horizons and learned methods that we could apply to the preparation of iGEM to help us design experiments more creatively and plan HP work more efficiently and impactably in the future. Next, they answered questions from other students in the form of booths, which deepened the communication between different teams. This exchange will let the team members fully understand the charm of synthetic biology and a wide range of application prospects!

2.2iGEM Greater Bay Area Industry-Academia-Research Forum (1st iGEM Greater Bay Area Industry-Academia-Research Forum)

On August 19, we, together with several iGEM teams from universities in the Guangdong-Hong Kong-Macao Greater Bay Area, as well as several professors and related enterprises, gathered at the Zhuhai campus of Beijing Normal University to participate in the first iGEM Guangdong-Hong Kong-Macao Greater Bay Area Synthetic Biology Industry-University-Research Forum. During this period, we listened to the introduction of many heads of pharmaceutical companies and learned about cutting-edge technologies such as stem cell therapy and liquid biopsy of cancer. Next, we communicated with a number of iGEM teams from Guangdong, Hong Kong, and Macao. In the in-depth conversations, we learned about many interesting projects and the rigorous experimental ideas behind them, and we learned how to do a good job in HP and the cooperation demands of biomedicine-related enterprises. On August 20, the second day of the exchange, we visited Tomson Bihealth, a well-known health products company, visited the production line of drugs, and conducted health tests, including glycosylation end product detection, bone density detection, and human substance concentration detection. Later, we came to Zhuhai University Institute of Science and Technology and saw the new foam cement developed by the UM team. By hand-weighing the new cement block and the traditional cement block of the same volume, we found that the new cement is significantly lighter than the traditional cement. The application of this cement greatly reduces the construction cost, and the load-bearing capacity is not inferior to traditional cement. From this example, we can fully feel the tremendous energy of industry-university-research cooperation. In this forum, our team has learned a lot, not only broadening our horizons but also inspiring new ideas for our follow-up competition, I believe that we will go further and further in the road of synthetic biology!

Our team arranges monthly exchanges with iGEM teams from other universities through online and offline means

2.3Up to now, there are a total of iGEM teams from Tsinghua University (THU), Lanzhou University (LZU), and Wuhan University (WHU-Antiphage). Not only to understand the research direction of other teams but also to state the research field of our team, in order to expand our influence. During this time, other teams gave us a lot of advice and help. For example, we reached a common concept of cooperation with the iGEM team of Lanzhou University, we helped them distribute questionnaires, and they gave us some hospital perception data of their affiliated hospitals. Our two teams also exchanged ideas about the work of HP, the clinical departments, and clinical directions so that the two sides could communicate with each other and improve the forms and programs of publicity. We learned a lot in lively discussions with several other teams. After that, we will better improve this work and let more people understand the charm of synthetic biology.

3. For education to public

3.1 Education for primary & secondary school

In the past year, we have not only made remarkable achievements in the field of science popularization but also deepened into primary schools, bringing valuable knowledge and inspiration to students. In order to integrate scientific knowledge into the education system, we actively seek cooperation with schools and educational institutions. We have established long-term partnerships with a number of primary and secondary schools to provide students with resources and support for science education. We have introduced synthetic biology to primary and secondary school students and preached about our topics, helping them understand the content more clearly and easily through games and lectures.

The following is our publicity experience in the field of volunteer teaching in primary schools:

With the joint efforts of our team members, we decided to bring the concepts of synthetic biology and microbial therapy into primary education. We believe that by introducing students to the concepts of science and innovation, we can stimulate their interest in the future and foster their creativity.

First, we contacted a local primary school and received support from the school. We have developed a series of lively and interesting lesson plans, including the basics of microbes, the fundamentals of synthetic biology, and an introduction to phage therapy. These courses include not only classroom instruction but also laboratory visits and interactive activities to ensure that students can fully understand and experience the joys of science.

In the classroom, we use lively teaching methods such as experimental demonstrations, scientific minigames, and interactive questions and answers. These methods not only arouse students' interest in the course content but also enhance their engagement. We also encourage students to ask questions and stimulate their spirit of inquiry.

Lab visits are an important part of the course. We took the students on a tour of our laboratory to demonstrate the specific applications and experimental processes of synthetic biology. Students developed a keen interest in laboratory equipment and operations, and some even expressed a desire to become scientists in the future.

In addition to classroom instruction and lab visits, we also had an interactive Q&A session with the students. The students actively asked questions and showed a strong curiosity about science and technology. We encourage them to think about future scientific developments and innovative applications.

During the volunteer teaching period, we not only pass on knowledge but also establish deep friendships with the students. We shared our research experiences and dreams, encouraging students to pursue their dreams and believe they can change the world.

Through this volunteer teaching activity in primary school, we not only spread scientific knowledge but also stimulated the interest and potential of students. We hope that these young minds will become scientists, engineers, and innovators in the future, contributing to the progress and development of society. We also want to continue to support primary education to open the doors of science to more students and let them see the infinite possibilities of knowledge.

This year, we not only made some achievements in the field of science popularization but also left a profound mark in the field of volunteer teaching in primary schools. This experience brought home to us the importance of education and the responsibility each of us has to contribute to the growth and future prosperity of the next generation. We will continue our efforts to spread science and knowledge to more people and work tirelessly to build a better future. During the summer vacation, we also hope to let more students come into contact with our team through an online volunteer teaching cloud, actively publicize synthetic biology and subject content, and continue to expand our influence.

3.2 Education for university students

For college students, we jointly organize large-scale charity sales with public welfare organizations on campus to expand the publicity of topics on campus. During the charity sale, we actively promoted knowledge related to synthetic biology and bacteria, and in the process of carrying out public welfare, the audience participating in the activity had a fuller understanding of the application and value of synthetic biology in life. At the same time, iGEM was brought into everyone's vision and won unanimous recognition and approval from many students. The promotion method of the booth is sand painting, and people can have a more intuitive understanding of the invisible microorganisms by hand-drawing the microbial images. At the same time, our team members customized postcards and canvas bags printed with iGEM and the team logo for distribution. More and more partners became interested in synthetic biology and iGEM and were willing to further understand the mystery of synthetic biology with us. During the charity sale, we also communicated with students from the BASIS China team, and the two teams will learn from each other and promote each other in the future, and jointly explore the beauty and possibility of synthetic biology.

In addition, we also cooperated with three other iGEM teams in the university to invite students and professors in the field of microbiology and synthetic biology to attend the seminar and get valuable suggestions by holding symposiums and forwarding posters. After the meeting, our teams exchanged with each other and deepened new insights on the follow-up experiment.

3.3 Popular science exhibition

Our team attaches great importance to the cultivation of synthetic biology concepts for children from an early age, so we came to Guangdong Science Center to spread the knowledge of synthetic biology to children.

We use posters and boards to educate kids about synthetic biology and the basics of our program, such as what bacteria, viruses, and phages are. To people's surprise, the children were very interested in the knowledge of life science. Many of them gathered around us to listen to our explanation. They listened carefully and actively asked us questions, and we answered them patiently.

Our activity lasted from 8 a.m. to 3 p.m., serving nearly a thousand children. Although this is a small amount in the total number of children, we have planted a small seed of synthetic biology in the hearts of these 1,000 children, and they will grow and continue to contribute to synthetic biology.

3.5 Online science popularization conference

In order to exchange and share the knowledge of synthetic biology with more people, we held an online science-sharing meeting with BNUZH-China and other teams.

We shared our knowledge about synthetic biology and our project with other teams and viewers online. The online audience is over 100 people, and in addition to the different teams, there are many friends who are interested in synthetic biology, and we introduce them, and accept their suggestions and inquiries. Synthetic biology and the related knowledge of the academy are expanded in a larger scope.

Sustainable Development

Antibiotic resistance is an increasingly serious threat to global public health, and a report released by the World Health Organization (WHO) has indicated that the era of "post-antibiotic" is approaching[1]. “A post-antibiotic era — in which common infections and minor injuries can kill — far from being an apocalyptic fantasy, is instead a very real possibility for the twenty-first century.” wrote Keiji Fukuda, WHO assistant director-general for health security[1]. According to an evaluation of drug-resistant infections in the Lancet, 4.95 million deaths worldwide in 2019 were related to antimicrobial resistance, with 1.2 million deaths directly attributed to antimicrobial resistance[2] (Reference figure 1). Since the discovery of penicillin, it was once thought that bacterial infections had been overcome with antibiotics. However, due to the increasing bacterial resistance caused by many factors, the impact of bacterial infections on public health is increasing day by day.

Among these strains, widely resistant Pseudomonas aeruginosa (PA) has posed serious challenges to public health worldwide. As one of the most common bacteria in hospital-acquired infections, it attacks the human host when normal immune defenses are disrupted, and is one of the key bacteria to guard against in the public health care system. In the United States, multidrug-resistant P. aeruginosa was responsible for 13% of severe healthcare-associated infections. Likewise, the European Center for Disease Prevention and Control reported in 2015 that 13.7% of PA isolates exhibit resistance to a minimum of three different antimicrobial groups[3]. According to data from the China Antimicrobial Resistance Surveillance System, the resistance rate of P. aeruginosa to carbapenems in 2021 was 17.7%. Indeed, in 2017, the World Health Organization (WHO) emphasized the urgency of the situation by categorizing carbapenem-resistant P. aeruginosa as a "critical" group for which new antibiotics were urgently required[4].

However, the development and production of new antibiotics are not actually sufficient to meet the new therapies needed to resist drug resistance, which means that the infection caused by these antibiotic-resistant bacteria poses a considerable threat to the incidence rate and mortality of the world.

Among all alternative therapy studies, bacteriophage therapy is one of the most promising methods. Bacteriophages, or simply phages, are viruses that have shown remarkable potential in combating drug-resistant bacteria. Their effectiveness stems from their unique mechanisms of action, which involve viral infection and subsequent bacterial lysis[5]. Additionally, they exhibit the ability to effectively control and combat infections associated with biofilms. Over the past few years, phage therapy has been successfully used to treat life-threatening multidrug-resistant bacterial infections, resulting in a large number of high-profile clinical case reports (Reference Table 1), and its efficacy in real-world clinical treatment and clinical adjuncts has begun to emerge. Furthermore, one of the significant advantages of bacteriophage therapy is its safety profile when it comes to mammals. A comprehensive review of the safety of bacteriophages in humans and animals has concluded that, based on current research results, the results of various routes of bacteriophage administration are usually safe and well tolerated[6].

For P. aeruginosa -targeting phages, many studies have shown their role in killing recalcitrant bacteria by hindering and disrupting biofilm formation. In terms of drug delivery, nebulization may be an effective and safe method. Animal experiments that have been reported demonstrate the efficacy and safety of inhaled phage therapy for combating P. aeruginosa infections[7]. The inhalation of phage powder resulted in a substantial reduction in the P. aeruginosa bacterial load within the lungs of mice (Reference figure 2). In addition, therapeutic approaches that combine phages with antibiotics also show significant promise, particularly in combating bacterial resistance. The synergistic effect of this phage-drug combination therapy makes it far more effective than single-drug treatment, providing a more attractive option for treating drug-resistant microbial infections[8].

Therefore, phage therapy has broad prospects in the foreseeable future and will have a positive impact on human health, human well-being, and sustainable development. In healthcare, phage therapy offers new treatment avenues for persistent bacterial infections to combat the growing global problem of multidrug-resistant bacteria. At the same time, this therapy reduces dependence on antibiotics and minimizes the proliferation of drug-resistant strains. In addition, phage therapy can also provide individualized treatment based on the specific circumstances of the infection, improving treatment effectiveness and reducing adverse reactions.

Concerning human well-being, containing the proliferation of multidrug-resistant bacteria can expedite patient recovery and enhance their quality of life. Furthermore, phage therapy holds the potential to alleviate the financial burden on the healthcare system and the public, as fewer patients with bacterial infections will necessitate prolonged hospital stays or costly treatments.

As is well known, the invention of antibiotics opened a new era in the treatment of bacterial diseases and saved countless lives. Nowadays, how to combat antibiotic resistance is also undoubtedly an important issue in extending human life and improving human health and well-being. The question of whether phage therapy can indeed serve as a beacon of hope for sustainable development hinges on ongoing exploration and research.

References

1.Reardon, S. WHO warns against 'post-antibiotic' era. Nature (2014).

2.Antimicrobial Resistance Collaborators (2022). Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet (London, England), 399(10325), 629–655.

3.Horcajada, J. P., Montero, M., Oliver, A., Sorlí, L., Luque, S., Gómez-Zorrilla, S., Benito, N., & Grau, S. (2019). Epidemiology and Treatment of Multidrug-Resistant and Extensively Drug-Resistant Pseudomonas aeruginosa Infections. Clinical microbiology reviews, 32(4), e00031-19.

Tacconelli, E., Carrara, E., Savoldi, A., Harbarth, S., Mendelson, M., Monnet, D. L., Pulcini, C., Kahlmeter, G., Kluytmans, J., Carmeli, Y., Ouellette, M., Outterson, K., Patel, J., Cavaleri, M., Cox, E. M., Houchens, C. R., Grayson, M. L., Hansen, P., Singh, N., Theuretzbacher, U., … WHO Pathogens Priority List Working Group (2018). Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. The Lancet. Infectious diseases, 18(3), 318–327.

5.Górski, A., Międzybrodzki, R., Węgrzyn, G., Jończyk-Matysiak, E., Borysowski, J., & Weber-Dąbrowska, B. (2020). Phage therapy: Current status and perspectives. Medicinal research reviews, 40(1), 459–463.

6.Liu, D., Van Belleghem, J. D., de Vries, C. R., Burgener, E., Chen, Q., Manasherob, R., Aronson, J. R., Amanatullah, D. F., Tamma, P. D., & Suh, G. A. (2021). The Safety and Toxicity of Phage Therapy: A Review of Animal and Clinical Studies. Viruses, 13(7), 1268.

7.Chang, R. Y. K., Chen, K., Wang, J., Wallin, M., Britton, W., Morales, S., Kutter, E., Li, J., & Chan, H. K. (2018). Proof-of-Principle Study in a Murine Lung Infection Model of Antipseudomonal Activity of Phage PEV20 in a Dry-Powder Formulation. Antimicrobial agents and chemotherapy, 62(2), e01714-17.

8.Uchiyama, J., Shigehisa, R., Nasukawa, T., Mizukami, K., Takemura-Uchiyama, I., Ujihara, T., Murakami, H., Imanishi, I., Nishifuji, K., Sakaguchi, M., & Matsuzaki, S. (2018). Piperacillin and ceftazidime produce the strongest synergistic phage-antibiotic effect in Pseudomonas aeruginosa. Archives of virology, 163(7), 1941–1948.

9.Singh, J., Yeoh, E., Fitzgerald, D. A., & Selvadurai, H. (2023). A systematic review on the use of bacteriophage in treating Staphylococcus aureus and Pseudomonas aeruginosa infections in cystic fibrosis. Paediatric respiratory reviews, S1526-0542(23)00045-3. Advance online publication.