Team:NUDT CHINA/Human Practices/Backup
Backup
Truly permeate people's life
Integration
Basical Crawler
Objectives:We wanted to know how other iGEM teams were doing since 2011 on diabetes related field, what
they have done or haven't done, and where we could explore further.
Main Part:We used a crawler approach to sort out those projects’ pages through keywords. Through manual
screening and sorting, we extracted the main content and HP activities of those projects. Based on that, we were able
to see how many projects are related to diabetes and what have they achieved.
Conclusion: Out of the 2045 iGEM team projects since 2011, only 0.9% were related to diabetes. Their HP
achievements include producing pamphlets, making documentaries, developing software, doing risk assessments, and
more. We learned from all these forms of human practices, and later constructed our own communication pathway with
the diabetic community.
Nurse Interview
Objectives:
With a basic understanding of the incidence of diabetes, we were eager to know the problems faced by patients with
diabetes and what we can do for them. Therefore, we had a nurse interview and visit.
Main Part:
Realizing that treatments of T2D are carried out directly by nurses, we contacted Guoxiu Li, head nurse of the
Department of General Medicine (former Gerontology) of the Second Affiliated Hospital of Chongqing Medical
University. With Ms. Li's consent, we conducted a nurse interview and hospital visit.
Ms. Li showed us the nursing work of diabetic patients and answered our questions about specific problems
arising from current treatments. The Second Affiliated Hospital of CMU also agreed to provide our team with more
treatment data of the patients for further analysis.
When we asked the nurses, who are in long-term first-hand contact with T2D patients, how they hope the treatment of
diabetes could be improved, they said that it would be nice if there were a convenient way of treatment that doesn't
require frequent medication. As a chronic disease, diabetes requires lifelong treatment, and the treatment process is
tedious, which brings physical and psychological burden to patients, many diabetics want simpler treatments that
ameliorate pain. If patients could be freed from the pain of frequent insulin injections and cumbersome treatment
procedures, they would cooperate with clinical treatments more actively.
Conclusion:
Currently, insulin together with some other insulin sensitivity improving medicines are the most widely applied
approach for the glycemia control of diabetic patients. However, constant and frequent insulin injection would bring
too much pain the patients. They are eager for a new way to treat diabetes.
The Status Quo Investigation of T2D
Objectives:
To understand the global status quo and the prevalence trend of diabetes.
Main Part:
According to the global report on diabetes from WHO (World Health Organization) in 2016, we found the prevalence of diabetes had significantly increased over the past decades, which alerted our team that diabetes has been more and more serious over the past decades. By currently public data, it is notable that there iare more than 352 million people around the globe at risk of developing such a chronic and painful disease, particularly in low – and middle-income countries. In China, around 10% of its population have diabetes, with nearly 60% exposed to the T2D related risk factors.
Figure 1
Figure 2
Conclusion:
Current data showed a significant increase of prevalence of diabetes in a global manner. These results highlighted the undiminished importance of developing therapeutic approaches for diabetic mellitus.
Reference:
Cho, N. H. et al. IDF Diabetes Atlas: Global estimates of diabetes prevalence for 2017 and projections for 2045. Diabetes Research & Clinical Practice 138, 271 (2018).
Clinical Interview
Objectives:To delve deeper into the issue, we would need to know what are some methods of treatment
and how are the treatments evaluated from an expert point of view. We were also interested in what existing ways to
control blood sugar in diabetics does the hospitals in our community offer. Therefore, in July and August, we
conducted diabetes-related interviews with relevant professional doctors both online and offline to improve our
subject.
Main Part:
We produced a survey that was filled out by 14 clinicians from different regions and different levels of hospitals,
including the associate chief physician of endocrinology, the chief physician of geriatrics, and the chief
physician of internal medicine. Below is a more detailed breakdown of the questions we asked.
After collecting the questionnaire responses, we sorted out these key takeaways:
1. Current characteristics and tendencies of diabetes
• Middle-aged, female, obesity
2. Evaluation of clinical approaches
• Oral medication/Insulin injections/GBP
• DPP-4 inhibitor (decreases glucagon levels)
• SLT2 (increases urine sugar excretion); rosiglitazone (increases insulin sensitivity)
3. Comparison of regional differences
• Treatments offered by township hospitals are simpler than those of city hospitals.
Conclusion: According to the clinicians’ responses, the elderly, female, and obese community make up most of the diabetic demographics, with no obvious trend of younger age. Diabetes treatment in the county and township is relatively simple. Currently, the main treatment is oral drug therapy and insulin injection, but there are other treatments, such as gastric bypass surgery. New drugs and treatment models are being developed in cutting-edge research, such as DPP-4 inhibitors that reduce glucagon, SLT2 that increases urine sugar excretion, and rosiglitazone that increases insulin sensitivity. Also, diabetes usually occurs with hyperglucagonemia. These conclusions were taken into consideration by our team while devising experiments.
The 22nd Scientific Meeting of the Chinese
Diabetes Society
Objectives:
In order to know more about the frontier research field of diabetes, our team sent representatives to attended
the 22nd national academic conference in Jiangsu held by the Chinese Diabetes Society on November 28, 2018.
Main Part:
During the 4-day conference, our members joined special seminars and hot discussions in various fields
related to diabetes. The conference allowed us to engage with well-known experts at home and abroad. We have
benefited a lot from the analysis of diabetes cases.
Our members also participated in the "Five-kilometer Health run," finishing the whole run with other runners.
It certainly was a refreshing experience since nobody is in lab clothing or writing papers in front of the
computer all way, but what’s more is that we feel the excitement of doing our part, together with other lab
workers and doctors, to appeal to the whole society to pay attention to the cause of diabetes prevention.
Conclusion:
Our attendees benefited from the inspiring lectures and academic exchanges on knowledge updating, and had the opportunity to learn about the latest developments in diabetes drugs and devices from the exhibition attached to the conference.
These results let us keep our eyes on the treatment of type 2 diabetes. Providing a way to cure type 2 diabetes must be valuable
2018 iGEM Project
Objectives: Noticing some improvements could still be made based on our last year's project, we thought
about how we could use the system we developed last year for glycemia controlling purposes.
Inspirations:
Diabetes Awareness Raising Program with
the Red Cross
Objectives:
Our goal is to spread knowledge on the treatment and prevention of diabetes and understand what challenges diabetic patients face in real life.
Main Part:
The previous online survey shows various degrees of misunderstanding on diabetes both in the elderly and the young, especially the elderly. Among the 524 valid questionnaires received by our team, only 12.4% of the respondents (65 persons) answered all questions correctly.
Considering our project aims to create a novel clinical application of type II diabetes mellitus (T2D), specifically by degrading GCGR to help decrease the blood glucose level, we believe it’s indispensable to conduct a popularization of diabetes in communities.
One member of our team, Yuhang Ye, is a member of the Red Cross. The Red Cross is one of the many local non-profit organizations that promote healthy lifestyles to the general public to prevent diabetes and other dietary diseases. Sharing the same goals, we collaborated with the local Red Cross. Through Ye’s contact, our group and the Red Cross jointly held the Diabetes Knowledge Popularization Lecture in the old people’s home of Changsha No.1 Social Welfare Institute on June 23rd.
Our team members handed out self-made brochures to the old, explained the mechanisms of diabetes and its treatment, and introduced our project, including an Q&A section at the end of the lecture.
Both young volunteers and the elderly at the Welfare Institute were interested in these topics that relate closely to everyday life. After the lecture, they eagerly asked questions about the lifestyle of diabetics, and we were more than happy to answer. Before leaving, the caretaker asked our contact information hoping for long-term communication on diabetic diets, believing our lectures would be more accessible to the elderly than those of clinicians.
The Red Cross in Changsha also decided that they would hold regular health knowledge popularization activities with NUDT_CHINA. We firmly believe that our project is beneficial to the community. We have future plans to set up a two-way communication website, on which not only we could guide the treatment and prevention of diabetes, but also diabetic patients can help us understand what particular challenges they face in real life.
After receiving overwhelming responses in follow-up visits in Changsha, we expanded our outreach and organized diabetes knowledge talks in Chongqing and Guangzhou.
Conclusion:
Following the previous on-line survey, we conducted a series of diabetes knowledge popularization lecture in several communities, formed a collaboration with the local Red Cross, and now have the goal to maintain and develop this two-way communication system through building websites.
These results let us keep our eyes on the treatment of type 2 diabetes. Providing a way to cure type 2 diabetes must be valuable.
Questionnaire on Measuring the General Public's Knowledge of Diabetes
Objectives:
Evaluate the general understanding of our society on knowledges related to diabetes.
Main Part:
Participants were asked to complete 7 True-or-False questions regarding the most fundamental knowledge about diabetes. 524 people were investigated by random sampling on the Internet. We categorized their ages into 4 distinctive groups: Juvenile, Young, Middle-Aged, and Aged. Among the 524 people surveyed, 47.3% are middle-aged, 16.4% are Juvenile, and 36.3% are over the age of 40. The gender ratio is roughly half-to-half.
After collecting and analyzing statistics, we found that only 65 (~8.5%) people answered every question correctly. Among them, 43 were from the youth group, and 18 were from the middle-aged group. The results show that middle-aged people and the young are most capable of distinguishing effective and ineffective methods in the treatment of diabetes. The aged and Juveniles groups, in contrast, are less familiar with the knowledge. This phenomenon may be caused by the lack of general diabetic education among juveniles and the misleading social superstition that is popular among the old.
Take the statement in question 6, for example: "Eating bitter melon can lower blood sugar levels." The correct answer is "False." However, only half of the juvenile group and low to 36.4% of the Aged group chose the correct answer. The theory that by merely eating bitter melons, one can reduce blood sugar levels currently exists only in folk remedies with no scientific research to prove. Some aged patients, who especially believe in those remedies, decide to eat bitter melon all day instead of taking medicine - This is not advisable.
Conclusion:
Regarding the results and the prevalence of diabetes among people in China, it seems surprising to us that 87.6% of the surveyed have misunderstandings on this significant public health problem. Therefore, we think it necessary to give lectures to popularize the appropriate way of curing diabetes and decided to make the old our major target audience.
Experts
Objectives:
With our preliminary data, we expected to receive more information on how we could improve our design. Hence, we discussed with several experts, showed them our preliminary data and ask for some more suggestions.
Main Part:
Our team had a holistic design of our project but was still searching for proof of our design’s feasibility and looking for aspirations. We decided to turn to experts in both the academic and the medical field. Through the contact of our team member Deng Yiman, we got in touch with experts Dr. Zheng Hongting and Dr. Xu Jing.
Figure1: interview with experts
I. Interview with Dr. Zheng Hongting
Deng Yiman: Hello, Dr. Zheng, please allow me to briefly introduce our project. Our team captured the issue of Type 2 diabetes (T2D), which is one of the biggest threats to human health. Previous studies have revealed the important role of glucagon in hyperglycemia in T2D patients. In our project, we demonstrated a design cell approach to control hyperglycemia by degrading the hepatic glucagon receptor (GCGR). We achieved this by controlling the GCGR degradation element with the CMV promoter. The Trim21 based proteasome system then performs GCGR degradation. This method provides a promising new treatment for T2D.
Figure2: Brief Introduction of Dr. Zheng Hongting
Deng Yiman: Our next plan is to test the design on primary cells. Do you think this is a viable method?
Dr. Zheng Hongting: Indeed, since the discovery of the glucagon receptor in 1971, it has been the focus of research in our field and is considered as a potential new target for hypoglycemic control. Moreover, GCGR knockout mice also showed significant improvement in glucose tolerance, but how to effectively reduce the expression of GCGR is worth further study. Your project to the degradation of GCGR provides a new solution, the in vitro results are exciting. However, the body’s internal mechanism is complicated, so you also need the results of ex vivo or in vitro, especially in animal disease model, to prove the effectiveness of the at the same time, also verify the security.
Deng Yiman: We use CMV as a promoter. What is your opinion on the feasibility of our project, and what advice do you have on our use of adenoviruses?
Dr. Zheng Hongting: By CMV promoter of words may cause systemic organizations GCGR widespread degradation; there may be a problem that should be considered: Although GCGR mainly express parts in the liver, it is also expressed in the kidney, adipose tissue, and the central system. The function of these sites is not entirely clear, and we do not know whether there are other physiological roles and functions yet. Would it be better to consider replacing CMV with liver-specific promoters such as TBG?
Deng Yiman: Thank you for your help and affirmation. We will continue our efforts!
II. Interview with Dr. Xu Jing
Deng Yiman: (*After introducing the team’s project) We use CMV as a promoter. Does this possibly cause hypoglycemia?
Dr. Xu Jing: Once the CMV promotor works in the body, it doesn’t stop. When it is overexpressed, GCGR is over degraded, and hypoglycemia is likely to occur. There are already examples of hypoglycemia occurring during the use of hypoglycemic agents, especially insulin secretion drugs such as sulfonylureas or glienides. In your team’s case, maybe consider replacing the CMV promoter with one that switches on and off at the right time, one that can achieve somewhat “intelligent” control.
Figure 3: Brief Introduction of Dr. Xu Jing
Deng Yiman: We hope that our product will ultimately face the public. Do you think our product carries clinical potential?
Dr. Xu Jing: Yes, in recent years, patients with diabetes due to obesity are getting younger, so there is a great market demand for diabetes treatment. Existing treatments include lifestyle interventions, drug interventions, and more. However, gastrointestinal reactions, weight gain, hypoglycemia, and other adverse reactions may occur during treatment. For example, oral medicine alpha-glucosidase inhibitors may cause nausea and vomiting. The relatively new drug, GLP-1, can reduce weight, but it works in the form of injections and can be painful. Your team’s product is designed to be easy to take in and unpainful to use, so it has clinical potential. I hope these ideas can be helpful to your team. Your project is very promising. Keep up the good work, and I would be happy to provide support if needed.
Deng Yiman: Thank you, Dr. Xu!
Conclusions:
In the interview, the professors we interviewed highly remark our projects as "open another window to control of blood glucose in a smart and intelligent way". They also provided important suggestions that we should further consider in future experiments:
• We should consider using more complex platform to challenge our circuit, eg. ex vivo or in vivo experiments;
• We should also consider the possibility of hypoglycemia caused by the constitutional expression of GCGR degrading system, tissue specificity should also be considered;
• We should think about another gene delivery method instead of liposome in vivo study.
2019 Chongqing iGEM Meeting
On July 20, the solstice of 2019, the first Chongqing iGEM exchange meeting was successfully held in the Army Military Medical University. In July, the mountain city Chongqing welcomed its first cool weather in nearly 60 years, and the friendship of igermers from all over China ignited the spark of synthetic biology.
Our team is giving a presentation
The National University of Defense Technology participated and presented the topic at the conference. Through the questions of other students and the comments of iGEM seniors, we realized that there are still many shortcomings in our project display and HP planning. The main problems are the following two. First, the form of the subject presentation is too singular, and the design of the PPT is not sufficiently concise. Therefore, we have improved the form of the subject and considered the use of the form of comics as one of the means of presentation of the subject matter. Second, the HP activities did not form a complete closed-loop with the experimental content, and the contribution to the content of the subject was not outstanding. Therefore, we have designed more links in the original activities to obtain external information, based on this analysis to guide the experiment. And in the follow-up event planning, we also designed more activities to improve our closed-loop further.
Additionally, we met the representative team of the Ocean University of China at the meetup, and we share our ideas about Human Practice with each other. We were pleasantly surprised to find that we have many similar ideas with OUC in the synthetic biology education sector. After a period of further communication, we have cooperated in this regard. Through this exchange meeting, all teams enhanced their friendship and also experienced the clash of new ideas and seeking of common progress among different designs.
Investigation on Transfection Methods
Purpose :
Because primary cells are relatively difficult to transfect, and cultured hepatocytes do not proliferate easily in a non-adherent state, culture of uniformly transfected hepatocytes by cell sorting or other purification methods is limited. Therefore, we investigated several transfection methods in the hope of finding a method suitable for us.
The following are the advantages and disadvantages of several commonly used methods of introducing exogenous genes into mouse primary liver cells：
1.Liposome carrier
The liposome vector method has been proven to be useful for transfection of primary cells, but experiments have shown that the number of transfected cells gradually decreases in EHS medium. Therefore, we believe that this method is not the best choice for transfection of primary hepatocytes.
Read the documentation for more information.
2. Electroporation
The data show that electroporation is indeed suitable for transfection of primary cells. However, the method of electrotransfection has many uncertainties, which cannot guarantee the quality of our transgenic primary liver cells.
3. Receptor mediated method
Receptor-mediated endocytic vesicles can be transported to lysosomes and easily degraded by lysosomes, resulting in short expression time and low expression efficiency of target genes.
Read the documentation for more information.
4. Calcium phosphate co-precipitation technique
In previous experiments, successful transfection of exogenous DNA with primary cell culture has been difficult to achieve. According to the literature reviewed, this method is inefficient among several methods of exogenous gene transfer, and the results obtained from repeated experiments are quite different, so it is not considered.
Read the documentation for more information.
5. Retroviruses
It was simple to construct, with a maximum capacity of 8kb of exogenous genes, and integrated into the host cell genome without viral protein expression. However, it can only infect mitotic cells, and its preparation titer in vitro is low, and its random integration may cause "insertive mutation".
6. Herpes simplex virus
Herps simplex virus (HSV) has a natural affinity for non-dividing cells, and the volume of exogenous genes was up to 30kb. In vitro, the titer was similar to that of adenovirus. However, during construction, it is difficult to remove the genes related to the lysed cells and it is highly toxic to the cells.
Read the documentation for more information.
7. Adenovirus
Adenovirus can transfer genes into aperiodic tumor cells, suggesting its potential for transfection in primary hepatocytes. It is one of the most reported viral vectors in gene therapy of hepatocellular carcinoma. In vitro preparation titer is larger, the capacity of loading exogenous genes is larger, the risk of insertion mutation is lower, the cytotoxicity can be greatly reduced through modification so as to make it harmless to cells.
According to the reference, scientists used adeno-related virus as a vector to transfer pdx-1 into the liver of diabetic mice, and obtained a long-lasting, non-toxic and efficient expression. Our experiments required a transfection method capable of dose-response studies, high-efficiency transfection, and simultaneous expression of multiple genes, and adenovirus was able to meet these requirements. Adenoviruses come from a wide range of sources and do not require high biological inhibition, and recombinant viruses have been shown to induce expression of hepate-related cells in large doses.
Consequence : Through comparison, we think that adenovirus is our best choice. It can be transfected efficiently, which can improve the experimental efficiency. It does not integrate into the genome of the host cell to avoid irreversible and lasting effects on the cell. It has a large dose range, which allows us to carry out multiple experiments and test the expression efficiency of transferred genes in many cases.
Talk to HQ
Objectives:
To inquire the safety concerns about using adenovirus and primary hepatocytes.
Main part:
Since adenovirus is usually used as the gene delivery tool for primary cells, our team tried using adenovirus in our project. Considering using adenovirus might bring some potential risks, we talked to iGEM safety hub to confirm that it’s safe enough to apply adenovirus in the lab.
First, we consulted the iGEM Safety Committee for the animal using form and safety check-in form. Here is our dialogue.
Moreover, though adenovirus has been widely used in other fields, we found that most iGEM teams were not quite familiar with such vector, not to mention the safety risks it may raise. Hence, our teammates made a brochure about adenovirus and how we could deal with the risks it brings, justifying that our work is responsible to the community.
Conclusion:
Under specific safety precautions, we are allowed to use adenovirus and purchased mouse primary hepatocyte in our project.
Collaboration
With CSU_CHINA, we talked about safety and efficiency of methods we have currently used to transport gene circuits into cells, and made detailed discussion on how to improve the efficiency and safety of these methods. CSU thought it would be possible to use transferrin to improve transfection efficiency on transferrin receptor enriched cells. Our team tested transferrin delivery efficiency with Invitrogen Lipofectamine™ 2000 Reagent in mouse primary hepatocytes and compared the transfection efficiency with regular adenovirus approach.
Disable the responsiveness of Bootstrap by fixing the width of the container and using the first grid system tier. A link to collaboration
Questionnaire on Public Acceptance of Adenoviruses for Diabetes Treatment
Objectives
To know the general public's acceptance of Adenovirus based gene therapy for diabetes treatment.
Main part:
We conducted a survey in the form of online questionnaire to evaluate to what extent adenovirus use in diabetes treatment is accepted by the public. We investigated 537 people by random sampling on the Internet. The sample proportion is also based on the age distribution in China and the transfection mechanism has been given in the form of a sketch on the first page of the questionnaire.
Our survey results show that virus therapy is poorly accepted by the public – despite more than half of our subjects support our idea, those who accept it themselves have not yet make up 30% of the population. The negative impression of the virus likely causes people to hesitate in exposing themselves to an unfamiliar therapy like virus therapy.
Based on its public impression that is not optimistic enough, we believe that follow-up science popularization is indispensable.
Our team would later address these issues by launching public engagement program trying to make them feel less concerned about trying new treatments through by providing fundamental knowledge on virus and gene therapy. We have achieved initial success by forming a collaboration with Southern Medical University Science Education Association who would actively support us in realizing further goals.
Conclusion:
Outdoor Popular Science Activities for Adenovirus
Objectives:
To let society accept our work.
Main part:
Based on its public impression that is not optimistic enough, we believe that follow-up science popularization is indispensable.
To further enhance the public's understanding and acceptance of gene editing and viral therapy, especially the adenoviruses transfection mechanism in our project, 45 science popularization volunteers of the Southern Medical Science Education Association and HP team members carried out related educational groups in Guangzhou. Aiming to increase the proportion of citizens who are willing to participate in public science education activities, we selected Yuexiu Park as a place for our popular science activities, where there is a large influx of people on the weekend, mainly consisting of old people and families with children.
We provided them with the Synthetic Biology brochure of the Ocean University of China and our cartoon cards, expecting to introduce the mechanism of synthetic biology and virus therapy vividly. We also designed interesting games to introduce the process of adenoviruses transfection, and thus unravel the public's misunderstanding of gene editing and virus therapy, and further enhance their interest in synthetic biology. We exchanged contacts with children and parents who showed interest in synthetic biology so that we could invite them to follow-up activities.
Conclusion:
With these HP work, the use of adenovirus not only promoted our experiments (in the lab) but also helps the population gained a more profound perspective about virus based gene therapy that it been tested in multiple clinical trials around the world. This is how human practices connect the lab and community as well as keep our project accountable to the world.