Human Practice

1. Overview

This year, RDFZ_China aims to use synthetic biology techniques to transfer formulated genes into E. coli so that they can help humans break down fat in the large intestine. Our Human Practice is divided into two main parts: understanding obesity-related issues and improving our product.

In the first major aspect, we used questionnaires to understand people's thoughts about obesity and their expectations of weight loss, then interviewed the masses and physicians about the disadvantages of diet pills and the acceptance of products produced using synthetic biology.

Second, improve the product aspect. We attended the July 17th 2022 Synthetic Biology Panel to upgrade the product. In the discussion with Prof. Jinqun Huang, we got biosafety-related suggestions and come up with a kill-switch design for our product. We also interviewed an obstetrician and gynecologist to learn from her about the safety of our product for mothers and babies. Finally, we interviewed Besunyen (the largest weight loss biotech company in China) and Shenzhen Escope Tech. Our product was recognized and the market for the application was again identified.

2.1 Inspiration and Background Research

In our daily school life, we noticed that the fast food place on campus were always crowded with students from all grades coming to buy high-calorie foods such as chicken and fries, and there was a constant flow of fast food restaurants in front of the school, and we learned that there was a lot of students and teachers with weight problems in the school. So we went to investigate the relationship between diet and many other factors and the physical condition of people to assess if this is a problem that needs to be solved.

Based on the questionnaire data, we targeted potential consumers to people in our surroundings and then conducted interviews. Their identities are middle-aged teachers and entrepreneurs.

We found that the causes of obesity are diverse, so we summarized the exogenous obesity factors of potential consumers. These factors included sedentary study and work, frequent business trips and social events, difficulty in allocating extra time and energy to control alcohol consumption and exercise; fast food leading to high triglycerides, high blood lipids and high cholesterol; easy sweating and fatigue and therefore more reluctance to exercise; sleep issues due to occupational and life stress.

We found that obesity problems may be related to negative psychology, so we investigated the psychological problems of these potential consumers due to their excess weight. Including discomfort with physical appearance, concerns about physical illness caused by obesity, anxiety caused by eating disorders and recurrent weight, hindrances and stereotypes in the office and in life from all aspects of society.

We found that modern and diverse approaches to weight loss vary from person to person, so we surveyed these potential consumers to find out what they had tried and how they felt about it. They had tried dieting and short workouts, but work schedules interfered largely with the program; they took traditional weight loss drugs orally and found them to be severely irritating to the intestinal mucosa; they consumed meal replacements and took appetite suppressants to reduce food intake. They use small molecule injections for diabetes to lower their blood sugar, but they are not as effective as they thought.

We investigated the acceptability of the program to these potential consumers who felt that there was a need for a research program from a lay group and believed that it could help people with mild obesity problems to improve the intestinal environment and metabolic problems, avoid overabsorption of fatty acids in the body, provide some weight loss, and create positive psychological intake. While long-term solutions to weight and body image problems require a combination of a healthy lifestyle and products.

Interviews identified the audience as mildly exogenously obesity, excluding people without basic needs: people with genetic obesity, secondary obesity with metabolic problems but with healthy living and drinking habits; special groups such as pregnant women who need to ensure maintain a high level of nutritional intake, with high level of progestational hormone, lead to weight above the normal range; and people who must use extreme weight loss techniques such as gastrectomy to address severe weight problems essentially are not suitable for the product.

The need for the product is clearly defined as effective, without significant side effects and convenient. Most of the common principles of traditional weight loss pills are to lower blood lipids; reduce appetite/stomach contraction, thus reducing eating; and contain laxative ingredients that promote bowel movements with some side effects, whereas probiotics do not produce significant side effects. Despite the negative effects, most people with a strong desire to lose weight prefer to invest in quick-acting drugs. The effect of taking probiotics in the short term is not as obvious as that of traditional drugs, and consideration needs to be given to how to balance production costs and product effectiveness.

2.1 Professional Surgeon Interview

To examine the feasibility of using synthetic biology to design anti-obesity drugs, we made interviews with professional surgeons from the third hospital of Peking University.

Professional Surgeon Interview To examine the feasibility of using synthetic biology to design anti-obesity drugs, we made interviews with professional surgeons from Peking University third hospital. Doctor Li told us the current methods all contain potential risks. The only permitted drug Orlistat has the side effect of low blood pressure, and intestinal diseases. Moreover, physical treatment such as liposuction might cause skin problems and immunity issues. Therefore, he inspired us the feasibility of using synthetic biology methods to minimize side effects during the process of losing weight.

3. July 17th 2022 Synthetic Biology Panel

The 2022 synthetic biology panel was held by Gansu microbiology association, which offers a chance for iGEM teams to meetup and share their experiences.
At the time, our project was designed in a way that only the transportation of exogenous long chain fatty acid (LCFA) was enhanced, and our experiment data regarding how well our E.coli can transport and metabolize LCFA was below our expectation. During the meetup, experts and members of other teams inquired about our experiment proceedings and experimental designs, and we were eager to seek for advice on improvements. One of the experts participating the conference suggest that we should research more about the integrated LCFA transport and metabolism pathway and seek for other targets to enhance the E.coli’s ability to absorb LCFA.

We then realize the bacterial cell may not be able to digest the increased amount of LCFA transported into the cell, therefore we read extensively to choose another suitable component that may improve the performance of our bacterial cell. We found 3 more proteins related to the LCFA metabolism, among which FadD, A protein responsible for the ligation of CoA group onto LCFA and facilitate the bacterial metabolism of LCFA was regarded as the most suitable candidate. After adjusting our design and overexpressing both FadL and FadD, promoting both the transportation and the metabolism of LCFA, our E.coli showed great improvement in its performance.

Overall, the conference was an enlightening experience where we gained inspirations to upgrade our design.

4. Interview with Mr. Huang's company Wei Zhi Jun

4.1 Why we visited:

As a probiotic project, we hope to learn about the products of other probiotic companies, including the disease they treat, their production, their difficulties, as well as the general environment of the probiotic industry, referring to and improving our weight loss probiotics. Mr.Huang Jinqun's Weizhijun company focuses on intestinal bacteria transplantation, by putting healthy people’s intestinal bacteria into capsules and giving them to patients to eat, it can improve the patient's intestinal environment, thus, treating the disease.

4.2 Introduction to Weizhijun Company:

Weizhijun is founded in 2017, it is a leading Al pharmaceutical company focusing on intestinal microecology therapy in China, having an advanced and complete microecology drug research and development platform. More and more studies have found that intestinal microbes have an important relationship with human health and disease occurrence. Therefore, Weizhijun hopes to introduce intestinal bacteria transplantation into the intestinal tract of patients to help them re-establish intestinal microecology and to treat and improve various diseases.

4.3 Communication:

Mr.Huang shared with us how his company uses intestinal bacteria transplantation to treat various diseases:
1. Tumor treatment
The problem of "Tumor Anti PD-1 Resistance" can be overcome by intestinal bacteria transplantation, and tumor treatment can be improved. Mr. Huang showed us the results of animal experiments and clinical trials. In the pictures, the tumors of mice and patients who used the transplanted bacteria were significantly smaller.

2. Improving Autism
Mr. Huang explained the experimental process of autism drugs. The experiment tested the improvement of autism and gastrointestinal symptoms in children by enema and oral capsules. Social behavior and stereotypic behavior of autistic mice were also used to detect improvement in social deficits. The experimental results show that intestinal bacteria transplantation has a significant effect on autism.

3. Treatment of Atopic Dermatitis
Mr.Huang mentioned that the hospital helped a woman completely improve functional constipation and atopic dermatitis through the bacterial flora transplant provided by a 28-year-old male health body.

4. Asthma, psoriasis, inflammatory enteritis, etc., can be improved by the same method.

Question: How do you view the current situation and prospect of E. coli weight loss therapy? What side effects does it have, and does it need to be accompanied by education about the dietary habits of product users?

Answer: Most of the probiotic foods and supplements on the market today that claim to have weight loss benefits don't do what they claim. I think E. coli weight loss therapy needs to be combined with relevant education to be effective in helping people lose weight. In treating metabolic diseases such as obesity, the role of probiotics is not entirely dominant, it’s not a miracle drug. Maintenance of the gut microbiota would be difficult and the results would be limited if it was not accompanied by beneficial dietary changes. Better weight loss results can only be achieved if the use of probiotics is accompanied by education of users to eat less and get more exercise. At the same time, it is also a question of whether weight loss probiotics can be maintained in the intestine for a long time. This may require constantly using your probiotic product.

After learning about our project, Mr. Huang Appreciated our design and the direction we chose, but also questioned our biosafety part. Although our bacterial cell E.coli Nissle 1917 has been commercially available with no toxicity, he warned us that it’s better to have a kill-switch system that can kill the bacterial cell when exiting the intestine, preventing unpredictable negative effects on the environment. After getting his advice, we realized our insufficient in our project design and started to research and develop a kill-switch system (please see more on the biosafety part).

4.4 What we learned:

The communication with Mr. Huang made us confirm that the direction of intestinal microorganisms is correct. It is a new target for medical development, and the entrance of intestinal probiotics can make a great contribution to the treatment of human diseases. At the same time, we note that more weight loss education for society needs to be combined, and a diet and exercise suggestion table should be added to our probiotic product, making people eat probiotics to promote weight loss while implementing the health plan provided by us to reinforce the weight loss effect. Additionally, a kill-switch system should be added to the design in order to prevent unpredictable negative effects of bacteria leakage.

5. Interview with an obstetrician-gynecologist Huiqun Wang

5.1 Why interview

Our weight loss products target people who are obese and overweight, but such a large group is very complex, with prople in different situations, such as ordinary obesity caused by diet, obesity caused by psychological imbalance, and inherited obesity genes. One of the most special groups is women who are obese due to childbirth. We have previously browsed many weight loss products and health supplements, most of which are marked as not suitable for pregnant women. To improve the safety of the products in the later stages of our project, we need to understand the factors of obesity in maternal and postpartum women, why some weight loss products are not suitable for them, and whether our products are suitable for pregnant and lactating women, so we interviewed the former obstetrician-gynecologist, now the editor-in-chief of the journal and the minimally invasive surgeon.

5.2 Communication

Q: What are the causes of obesity in postpartum women?
A: Women become obese during pregnancy and after childbirth. When a woman is pregnant, the placenta in the body and amniotic fluid are at least 30 kg. In addition, to ensure the healthy development of the fetus, pregnant women will increase nutrient intake, if not well controlled, it will lead to excess nutrition, of which fat is almost grown in pregnant women. As a result, two-thirds of pregnant women will suffer from obesity. After giving birth, women do not naturally become thinner. Coupled with the need for breastfeeding, regular exercise cannot be guaranteed, and the weight of the body progesterone is high in the body, and its weight will not change much within at least one year. Usually, postpartum women are the fattest, after which it is only possible to maintain the status quo, not to get fatter, and the average person will recover in five or six years. Of course, there are exceptions, such as people who deliberately lose weight, and the means used include self-loss, non-feeding, external treatment, etc.

Q: How much are postpartum women willing to lose weight?
A: Some people will have the idea of losing weight after giving birth, but the difficulty of losing weight is different to different people, so the psychological feelings produced are also different. Most people will not feel uncomfortable with their body after childbirth, because pregnancy is fatter than after childbirth, and because the physical exertion during confinement is also large, so few people deliberately lose weight.

Q: After listening to our introduction to the project products, what is the acceptance and suggestion?
A: China now requires mothers to breastfeed exclusively for 6 months after giving birth, which requires mothers to eat nutrients to ensure the development of their children, so weight loss is impossible. In particular, some breast milk is still pale after rest, which means a lack of nutrition, so mothers especially need to ingest nutrients. If fat (nutrition) is broken down, the quality of breast milk is not good, which will affect breastfeeding and the development of the baby. This results in maternal obesity primarily. Postpartum mothers have a very healthy diet. While such a diet is unlikely to cause rapid weight loss in the short term, it can sustain weight loss.

At the same time, some people use gastrectomy (the fundus and gastric body are cut off so that the absorption area and storage area of the stomach are smaller) to achieve rapid weight loss. Since some women can't get pregnant because they're obese excessively, this surgery helps with ovulation. The surgery also applies to metabolic syndrome. Since this type of person needs fast and effective weight loss methods and can be cured in a short period, our weight loss products appear superfluous in front of this type of person.

5.3 Conclusion

The doctor agrees that our probiotics have no adverse effects on pregnant women, but during breastfeeding, mothers need to consume fat and nutrients, so weight loss itself is not advisable for them. This is possible for those who want to keep weight as they are without accumulating too much fat.

Therefore, we conclude that pregnant or lactating women should avoid using our products as much as possible. After lactation, the mother gradually accepts probiotics.

We also found that, in fact, potential people our probiotic weight loss product is not suitable for are still need us to explore, which not only improves the vigilance of our product release, but also improves the safety of our products to a certain extent.

6. Interview with Mr.Hong Yaowu from Besunyen Company

We visited the factory and laboratory of the largest weight loss biotechnology company in China, Besunyen Company, then we interviewed its Vice President, Hong Yaowu. Besunyen Company is the leading brand in the health care tea industry in China, enjoying a clear advantage and industry position, and also gaining recognition and popularity among young people in China. Besunyen produces products similar to ours that focus on human intestinal health and weight loss.

We first learned about Besunyen's corporate culture and concluded that one of the reasons for Besunyen's success is that it sees business opportunities and market demand, persistently researches a product, and conducts live webcasts to promote it in conjunction with the development of the times.

We visited Besunyen's factory and laboratory. We learned that there are many difficulties in moving from the lab to the factory for mass production, such as industrial, equipment, environmental, regulatory verification (Good Manufacture Practice), and financial issues.

In the interview with Vice President Hong Youwu, he first mentioned that there is a great market demand in China with a high obesity rate and low age. There is also a diversity of weight loss-related products and methods. However, weight loss should not rely only on medicines or health products but should be combined with exercise and a proper diet (dietary structure).
Secondly, we can learn from the fact that Besunyen Company has adapted to the needs of consumers by adopting group research on different consumers (eg participating in college advertising festivals), and community activities. And based on the results of the research to improve existing products or develop new products. The ideas and values of the product cannot be forgotten at this stage: scientific approach, modern health concepts, and society facing the obesity problem squarely. This helped us to further define the market size and sales positioning and gave us a business model that we can learn from.
He then re-emphasized the importance of product safety and offered suggestions when producing it. This product must be manufactured, stored, and transported in a way that the practices are in line with the regulations in order to ensure the safety of the product. He hoped that the cost of our products can be controlled so that we can achieve a larger audience and advantage in the future sales process (when we visited pharmacies before, we found a SHOWYOO stick probiotic whey, which was taken off the shelves because of the high price of buying fewer people).
Finally, he approved our project. Focusing on human health issues, and this green project can be accepted by the public. The acceptance of synthetic biology products has also increased with the rise in consumer demand. It is in line with the social trend and acceptance range as new technology. He showed his interest in our project.

7. Interview with Mr.Liu from Shenzhen Escope Tech Company

7.1 Why we visited?

At the time our product design is basically finished, it’s time for us to visit some business successful companies and get advice for our own product’s marketing. We contacted Mr.Liu Yuqi from Shenzhen Escope Tech as this company is renowned in the biomedical field. We received a reply that they are willing to communicate with us and offer us suggestions on business and some precautions.

7.2 Introduction to Shenzhen Escope Tech:

Shenzhen Escope Tech Co., LTD. is founded by professors and engineers who returned from Europe and America. Their direction is to use high technology to facilitate the medical field. At present, the company's product positioning is coronary artery analysis, heart disease function evaluation, intracranial artery analysis, brain function evaluation, and peripheral artery analysis five major fields.

7.3 Question & Answer:

1.Q: Which kind of functional product route do you think should we follow if our weight loss probiotics go into the market?
A: There are three ways: drugs, healthcare products, and food. Although the sales cost of drugs is low, since people will buy them without more need for advertising, the time and money cost of going through the drug marketing process is too large. Healthcare products are between drugs and food, with medium investment and medium harvest. Going through the process is simple for food, but the cost of publicity and promotion is high. I suggest you make your product a healthcare product so that the require for time and money are not high, and it can win the trust of the public. A very important part of weight loss products is to let the consumers believe your effects. Healthcare products’ efficacy is audited, which strengthened their credibility.

2.Q: What do you think about the future of our weight loss probiotics?
A: The market for weight loss products is very large, as long as you are safe and effective. Pricing can set two price systems, middle and high, because our target population is not low consumer groups. The difference between your probiotics and other weight loss probiotics is that they are natural bacterial cells, while yours has been genetically modified, which can have a more significant effect and can be used to convince customers. There is no problem in widespread selling as long as you ensure your product positioning, costs, legal issues, and target groups.

3. Q: What do you think about the relationship between obesity and cardiovascular disease?
A: Although obesity is not necessarily the direct cause of cardiovascular disease, they must be positively correlated. In other words, obesity is one of the causes of cardiovascular disease. That’s why it's important for cardiovascular physicians to ask their patients to control their weight.

7.4 What we learned:

The interview with Shenzhen Escope Tech is successful and insightful. We confirmed that our product’s position is a healthcare product which increases our product’s credibility. We paid additional attention to the legal issues around probiotics and ensured legal safety. Targeted groups we have already stated in the former part of potential consumers. Finally, we gained recognition from Mr. Liu through both a businessman's lens and a cardiovascular disease expert's lens.