1. Problem Discovering

Hepatitis B is an infectious disease caused by the hepatitis B virus (HBV). The annual number of deaths caused by hepatitis B infection has surpassed those caused by tuberculosis, HIV, and malaria, making it a significant public health problem that severely endangers human health. For example, there were 254 million people infected with hepatitis B worldwide in 2022. It is estimated that about 1.2 million people will be newly infected with hepatitis B every year, 1.1 million people will die of related diseases every year, and about 2900 people will die every day on average. (乙型肝炎 (who.int)) Chronic hepatitis B infection can lead to life-threatening conditions such as liver fibrosis, cirrhosis, and liver cancer.

China bears one of the highest burdens of HBV infection worldwide. According to a review report in The Lancet Gastroenterology and Hepatology, In 2022, the HBV prevalence rate in China was 5.6%, compared to the global prevalence rate of 3.2%. The proportions of patients with primary liver cancer and cirrhosis caused by HBV infection in China are 80% and 60%, respectively, significantly higher than the global incidence rates of 45% and 30%. The mortality rate of HBV related liver disease also accounts for 30% of the global HBV infection mortality rate.

In this regard, The World Health Organization (WHO) has set the goal of eliminating hepatitis B by 2030, emphasizing the need to improve the treatment rates of hepatitis B patients, reduce new cases, and calling on all countries to take proactive measures to combat viral hepatitis. In order to cope with the severe situation of hepatitis B infection in China, we proposed to comprehensively strengthen publicity and education, strengthen comprehensive intervention, strengthen referral and standardized treatment, strengthen drug supply, strengthen information management, increase detection efforts, implement medical insurance policy, and limit new infections of hepatitis B to the maximum extent. 中国肝炎防治基金会 (cfhpc.cn) China will be a major contributor to the goal of eliminating HBV, a threat to public health, by 2030. The criteria for HBV cure are: undetectable serum HBsAg, clearance of intrahepatic and serum HBVDNA (including intrahepatic cccDNA and integrated HBV-DNA), sustained positive serum anti-HBc, with or without the presence of anti-HBs.

We first learned about hepatitis B from the perspective of modern medicine in 1960, but it has been difficult to overcome for a long time. The main challenge is that existing treatments cannot completely cure hepatitis B. Currently, the most commonly used hepatitis B treatments, such as tenofovir and entecavir, only control the disease by inhibiting viral replication and cannot completely eliminate the DNA integrated into the host genome or the covalently closed circular DNA (cccDNA) that drives HBV replication and antigen expression. So the patients need to take a lifelong course of drugs to ameliorate liver injury. The World Health Organization has determined the clearance of serum HBsAg and HBV DNA as the latest "complete cure" standard for hepatitis B. For lack of specific targeted drugs that can clear intracellular cccDNA and integrate DNA into the host genome currently, even if serum HBsAg and HBV-DNA can be controlled, it is still difficult to achieve complete cure. Therefore, we need a drug that can accurately and targetedly cleave cccDNA.

2. Stakeholders' Map

Our primary focus is on people, so we need to gather information about the stakeholders involved in the project, including liver disease patients, research team members, healthcare professionals, pharmaceutical companies, medical insurance institutions, public health departments, government agencies, and investors. We primarily focus on the first four groups: what are their urgent needs, and what improvements do they hope to see? These true social needs and suggestions of these groups are crucial to the underlying construction of our project.

In order to protect patient privacy, we make full use of online resources to collect relevant information. We have learned that most patients with hepatitis B need antiviral drugs and related nursing drugs for a long time, and only a small percentage can achieve temporary cure of chronic hepatitis B. And during the treatment process, medication should not be easily discontinued, and adjuvant therapy drugs should be taken. Except for drug demand, we also focus on the mental health of patients. Most hepatitis B patients and carriers have difficulties with interpersonal communication, marriage, and employment. Their psychological pressure really cannot be ignored.

3. Project Feasibility

To enhance the design of our project, we seek input from stakeholders and experts from various fields, while also taking a comprehensive approach to understand the macro environment, including current policies, international concerns, drug consumption levels, technological development, and social values.

Support for Gene Editing Therapy:

3.1 According to the 14th Five-Year Plan and the 2035 Long-Range Objectives Outline, strategic emerging industries, including the cell and gene therapy industry, are expected to contribute over 17% of GDP.

3.2 The Shanghai Municipal Science and Technology Commission, Municipal Economic and Information Commission, and Municipal Health Commission jointly issued the "Action Plan for Promoting Gene Therapy Technology Innovation and Industrial Development in Shanghai (2023-2025)". It provides up to 30 million yuan as financial support for gene therapy products that have undergone clinical trials and obtained market approval in China. At the same time, gene therapy companies are encouraged to list on domestic and foreign capital markets such as the Science and Technology Innovation Board, providing financial support.

3.3 In the "Guiding Catalogue for Industrial Structure Adjustment (2024 Edition)" released by the National Development and Reform Commission, cell culture, gene therapy drugs, cell therapy drugs, and high-end intelligent pharmaceutical equipment are included in the encouraged industry catalogue, which will be implemented from February 1, 2024.

WHO 2030 Global Strategy:

The World Health Organization (WHO) has defined the goal of eliminating hepatitis B by 2030 and called on all countries to take positive measures to deal with viral hepatitis. Therefore, China has put forward the Guidelines for the Prevention and Treatment of Chronic Hepatitis B (2022 Edition), the Expert Consensus on the Whole Population Management of Hepatitis B (2023) and other improved treatment systems to reduce the infection rate and mortality rate of chronic hepatitis B.

Large Painent Base:

In 2002, the number of hepatitis B infected people worldwide reached 254 million, about 1.2 million people were newly infected with hepatitis B every year, 1.1 million people died of related diseases every year, and about 2900 people died every day on average. The huge number of infections represents the huge market of hepatitis B drugs.

Strong product competitiveness:

Our final product aims to eliminate hepatitis B virus in human body from the root, which is different from the inhibition of hepatitis B virus replication by market drugs. This is our biggest advantage.

Distinct Listing Risk:

Technological maturity and safety issues: The true application of CRISPR gene editing technology in the human body requires stable and accurate effects as well as precise control, all of which need to be further validated and improved.

Cost and accessibility: The development and production costs of CRISPR gene edited drugs are relatively high, which may lead to high drug prices and affect patient accessibility and affordability. Therefore, it is necessary to minimize costs during design and research and development, and coordinate with regional policies to make it as comprehensive and accessible as possible.

Long term effect and risk monitoring: the long-term effect and potential risk of CRISPR treatment of hepatitis B still need long-term follow-up research, which may affect the long-term benefits of the market and the trust of customers.

Patient education and acceptance: Patients and healthcare professionals may need time to understand and accept new technologies, which may affect the market penetration rate of new drugs.

Eliminating hepatitis B:

There are multiple societal demands for eliminating hepatitis B at various levels.

Public health needs:

Hepatitis B is a serious global public health problem, especially in China, where a large number of hepatitis B infected people pose a major threat to public health. At the same time, hepatitis B is also a global problem that needs the cooperation of the international community to solve together.

Economic development needs:

The treatment and prevention of hepatitis B require significant medical resources, which increases the burden on the healthcare system and places economic pressure on patients’ families. By eliminating hepatitis B, we can reduce the expenditure of medical expenses and improve the utilization efficiency of social resources.

Demand for scientific and technological progress:

With the progress of medical science and technology, especially the development of our gene editing technology such as CRISPR, new possibilities have been provided for the treatment of hepatitis B. The society has high expectations for these new technologies and hopes to thoroughly solve the problem of hepatitis B through scientific and technological progress.

Acceptance of Gene Editing:

The public's acceptance of gene editing technology is influenced by various factors, including the level of understanding of the technology, cultural background, religious beliefs, ethical considerations, and potential medical benefits. Therefore, we use survey questionnaires to investigate the public and analyze their acceptance and understanding of gene editing technology.

Maintenance of Social Justice:

Hepatitis B patients may suffer from discrimination and exclusion in society, which affects their mental health and even leads to severe social problems. By eliminating hepatitis B, we can try our best to eliminate social discrimination and promote social harmony. We take the universality of drugs into account for reducing the occurrence of unfair issues.

Problems in Current Therapies:

Current treatment drugs cannot achieve complete cure, and there are issues such as lifelong medication, drug resistance, and side effects. Some patients have insufficient understanding of hepatitis B treatment, which leads to poor treatment compliance and affects the treatment effect.

Technical Feasibility:

CRISPR Cas nucleases have demonstrated excellent capabilities in multiple fields. And it has achieved an ideal breakthrough in the treatment of hepatitis B. Even if there is still a distance from complete realization, the future is promising. During the project, we continuously explore and consult with external parties to improve the project while verifying its feasibility.

Technical Advantage:

Our project innovatively proposes new methods to treat hepatitis B, aiming to achieve accurate targeting and efficient elimination of hepatitis B virus in the human body. Combination therapy has great potential-it can be used in combination with other treatment methods such as antiviral drugs and immunomodulators to improve treatment effectiveness and ultimately achieve complete cure.

A perfect project requires rigorous design and precise thinking. We gather information from stakeholders and gain experience from predecessors.

In the process of seeking help and consultation, we have established a completed project framework that takes into account product safety, efficiency, and price. We have achieved theoretical perfection and completion in the design of wet and dry experiments, continuously expanded project boundaries, and improved commercialization to achieve the feasibility and perfection of the entire project.

4. Human Practice seeks external assistance, consultation and interviews

4.1 Dr. Yunkuan Zhao

Zhao Yunkuan, Deputy Chief Physician. Beneficial exploration has been made in the early diagnosis and treatment of severe hepatitis. In Jiaodong, he took the lead in trying out new technology of nucleoside anti hepatitis B drugs to treat subacute severe hepatitis B and hepatitis B cirrhosis, and accumulated rich clinical experience. Over the years, he has written articles such as "8 Cases of Chronic Severe Hepatitis Caused by Medical Intervention".

Action: Our members visited Qixia People's Hospital in their hometown during the holiday, and found Dr. Zhao there for consultation on the real treatment dilemma of hepatitis B, to verify the credibility and authenticity of the background collected data.

Feedback: Dr. Zhao, who is deeply involved in the clinical treatment of hepatitis B, noted that most patients cannot achieve a clinical cure for chronic hepatitis B and need lifelong therapy. The price of hepatitis B treatment related drugs has been reduced through medical insurance, and the burden of taking drugs to control the disease has been greatly reduced. However, lifelong medication, the side effects it causes and irreversible damage to physical health always have an impact on the lifelong happiness of patients.

Harvest: This consultation verified the credibility and authenticity of the information collected in the early stage, enabling us to think about the significance of the project on the ground.

4.2 Dr. Zhiying Chen

Chief Physician, Master's Supervisor at China Medical University, Master's Supervisor at Dalian Medical University, and part-time Associate Professor at Dalian Medical University, Member of the Digestive Immunology Professional Committee of Liaoning Immunological Society, Member of the Elderly Immunology Branch of Liaoning Immunological Society, and Member of the Inflammatory Bowel Disease Professional Committee of Liaoning Integrated Traditional Chinese and Western Medicine Society. Engaged in clinical work in gastroenterology for a long time, currently mainly focusing on clinical and scientific research on liver diseases.

Action: On line, we sent Dr. Chen consultations on the actual situation of hepatitis B treatment, technical feasibility of the project, practical considerations for patients, future vision and other contents, and received exact responses.

Feedback: Dr. Chen expressed her support for our project, believing it has great potential to achieve a complete cure for hepatitis B, but noted that significant effort is still needed to reach this goal. In terms of experimental design, challenges may exist in off-target issues and gene editing delivery efficiency issues. From the perspective of the patients, they hope to consider first the treatment effect, second the safety, and third the treatment cost.

Doctor Chen made an evaluation on the current treatment status: It is still difficult to achieve the ideal negative rate of HBeAg and HBsAg with the current use of antiviral drugs for hepatitis B. The large number of hepatitis B patients, the low treatment rate, and the limited treatment methods still pose many challenges to the beautiful vision of hepatitis B prevention and control in China.

Harvest: Through consulting Dr. Chen and conducting our background research, we have determined that safety and effectiveness will be key considerations in our project, which is reflected in the design and results of dry and wet experiments. And in the subsequent business plan, systematic consideration was given to issues such as product prices and treatment costs.

4.3 Dr. Jinghua Sun

Director, chief physician, doctoral supervisor, and master's supervisor. The Youth Committee of the Oncology Branch of Liaoning Medical Association, a member of the Tumor Prevention and Control Special Committee of Liaoning Preventive Medicine Association, and a member of the Tumor Immunology Special Committee of Liaoning Immunological Society. She masters the standardized and personalized advanced diagnosis and treatment techniques for malignant tumors, especially digestive system tumors, as well as the regulatory mechanisms of tumor occurrence and development.

Action: We chose the Second Affiliated Hospital of Dalian Medical University nearby and conducted interviews and consultations with authoritative doctors in the oncology department.

Feedback: Dr. Sun stated that our project is theoretically sound and feasible. Chen believes that the CRISPR technology we use may play a significant role in the treatment of other diseases. As an oncologist, Chen proposed that our project can be used as the primary prevention of liver cancer, not only for the complete cure of chronic hepatitis B, but also to prevent liver cancer and other diseases caused by hepatitis B. In terms of project products, Dr. Sun emphasized that drugs used on humans must be able to distinguish between ‘friend gene’ and ‘foe gene’, since safety is of utmost importance in this case.

Harvest: Dr. Chen showed us a deeper perspective of thinking. She suggest that we expand the scope of the project, not only serve the treatment of hepatitis B. but also provide value for the prevention of liver cancer. In the interview, the importance of technical security was repeatedly discussed. Therefore, safe gene editing should always be considered in project design, operation, and implementation.

4.4 Boteng Biology

Suzhou Boteng Biopharmaceutical Co., Ltd. is a CDMO (Contract Research and Production Organization) service provider specializing in the field of gene and cell therapy. The service scope of Boteng Biotechnology covers different stages of drug production, from early research, clinical trials initiated by researchers, New Drug Clinical Trial Applications (IND), registered clinical trials to commercial production. The company provides CDMO services including plasmids, viral vectors, and cell therapy products to support customers in accelerating the process of drug research and transformation.

Action: Project members collectively visited Suzhou Boteng Biopharmaceutical Co., Ltd., visited the product production plant, and exchanged project information with staff to obtain relevant information on market direction and experience.

Feedback: During the communication, the staff affirmed the significance and feasibility of our project, stating that it is closely related to some of the company's projects. They believe that after the project is implemented, it is possible to establish a connection with the company and collaborate with them to produce products. We have added contact information with the staff, which is expected to lead to cooperation in the future. In terms of market development, he shared with us the market development of gene editing tools. Meanwhile, the suggested technical suggestions have brought us new ideas.

Harvest: In this exchange, they showed us the current situation and future development trends of the gene editing drug market in terms of product presentation and market analysis, which will help us to seek benefits and avoid disadvantages in the commercialization process. In terms of technology, we designed the concept of introducing mutations and screening in CRISPR Cas nucleases will be considered and selectively applied to project design.

4.5 Hangzhou Juyuan Biotechnology Co., Ltd.

Hangzhou Juyuan Biotechnology Co., Ltd. is a high-tech enterprise located in Hangzhou, focusing on research and production in the field of biotechnology. The company's business scope includes technical services, technology development, technology consulting, technology exchange, technology transfer, technology promotion, and medical research.

Action: Our members got in touch with the engineer of the company online, who once worked in Zhengda Tianqing Pharmaceutical Group, the manufacturer of domestic hepatitis B treatment drug "entecavir". We communicated with them about the project, conducted interviews and consultations on treatment drugs, drug markets, and other related issues.

Feedback: The engineers of the group think that our project is "a real blessing that can benefit hundreds of millions of hepatitis B patients". Currently, the drugs used for hepatitis B treatment in China can only inhibit the replication of the hepatitis B virus. The commonly used "entecavir" costs 16.5 yuan per week after governmental centralized procurement. Although the cost of the drug is relatively low, patients' demand for a cure has intensified due to the improvement of living standards. This demand has given us great motivation to overcome the difficulties in experimentation and completion.

Harvest: The current market needs to be able to replace inefficient therapeutic drugs, and there is a huge demand for projects in the market. The market of hepatitis B therapeutic drugs is as broad as that of gene editing drugs.

4.6 Amy Integrity

Amy Integrity Biopharmaceutical Co., Ltd., founded in September 1993, is one of the earliest high-tech enterprises in China specializing in vaccine production, research and development, and sales. The recombinant hepatitis B vaccine (Hansenula yeast) adopts the domestically first and internationally advanced recombinant Hansenula yeast expression technology platform, which has the characteristics of high yield and high antigen expression purity. It has reliable safety and is suitable for a wider range of vaccination populations, and enjoys a very high reputation and influence in the market.

Action: After getting in touch with the company, we participate in on-site cognitive practice activities, and discuss issues related to hepatitis B vaccine and hepatitis B treatment with on-site staff.

Feedback: We communicated with the staff about the project and showed them the beautiful vision of complete cure of chronic hepatitis B. They affirmed the feasibility of our project ideas and thought that our project would bring hope to many hepatitis B patients.

The staff of Amy Integrity told us about Amy's arduous path in drug development, production, and product sales, demonstrating the standardized vaccine production process and technology platform, so they really understand the efforts we have paid. During the exchange and learning, we particularly noticed that drug production must strictly comply with national regulations. This is crucial for commercial production.

Harvest: This exchange has provided us with a warning for our future commercialization. And it reminds us that the drugs used in the human body cannot tolerate any mistakes, which also prompts us to achieve more rigorous experimental design and perfect drug products.

4.7 Haotian Guo

Dr. Guo Haotian, Marie Curie Scholar, CEO&co-founder of Ailurus Biotech, PhD from the University of Paris, former researcher at the Paris Interdisciplinary Research Institute, and visiting scholar at the MIT Department of Physics; 10 years of interdisciplinary research experience, first and corresponding author of multiple academic papers.

Action: We have invited Dr. Guo Hao to have a discussion and exchange with all project members, providing constructive suggestions and guidance on issues such as the project narrative framework and scope of work.

Feedback: Dr. Guo Haotian has provided suggestions for modifying the overall framework and work details of our project. In the dry and wet experiments, Guo suggested that we reduce the volume of work and focus on the core issues to complete the project in a more orderly manner. He stated that we need to focus on "Therapeutic" track to avoid deviation and repairing the narrative framework. This provides great guidance and assistance for our overall experiment and project presentation.

Harvest: Following the guidance of Senior Guo Haotian, we reduce the excess workload of the project and focus on the main pain points. We have gained a lot in project narrative skills and presentation structure.

4.8 Hangzhou Dixiang Industrial Co., Ltd

Hangzhou Dixiang Industrial Co., Ltd. focuses on the research, development, and application of advanced nanobiotechnology and liquid biopsy technology. The company has established extensive and in-depth cooperation with numerous top universities, hospitals, and research institutes in China, jointly undertaking national and provincial key research and development projects and special projects.

Action: Our members have contacted Mr. Zeng and the R&D department personnel of Hangzhou Dixiang Industrial Co., Ltd. for offline communication and consultation.

Feedback: After learning about our project, Zeng always believed that the technology used in our project also has great potential in other aspects, and regarded our CRISPR technology as a widely applicable tool, which expanded the boundaries of our project. Mr. Zeng believes that the incidence of hepatitis B in China has shown an overall downward trend, and it is feasible for us to focus on the treatment of patients who have already developed the disease. At the same time, we are advised to pay attention to the universal applicability of our project products, which is mainly reflected in drug prices and public acceptance.

Harvest: We expand the boundaries of the project- The technology we use can be modified and used for the treatment of other genetic diseases as a 'tool'. We have considered the issue of universal applicability of drugs in our business plan. We conducted a social survey on the public's acceptance of gene edited drugs and obtained answers from the public acceptance level.

4.9 Professor Fang Bai

Bai Fang, Professor, Researcher, and Doctoral Supervisor at Shanghai-Tech University. Her primary research interests are exploring and designing new drug design methods by combining computer-aided drug design, theoretical biophysics, and the use of molecular biology and cell biology methods. And now she is really experienced in developing novel drug design methods in combination with artificial intelligence algorithms and applying these methods to conduct research on the design of candidate therapeutic agents for major diseases, such as high-incidence cancers and major infectious diseases.

Action: We contacted Professor Bai Fang and requested assistance on experimental design and direction through online Tencent Meeting.

Feedback: Professor Bai Fang affirmed the feasibility of obtaining high-precision, high-activity targeted therapy through CRISPR-based directed evolution. Professor Bai Fang provided guidance and corrections for a series of issues we encountered during the experiment. These included operational details for positive prediction of wild-type MAD7, feasibility of building a Cas9 data model, exploration of the issue of small data volume, the relationship between mutation sites and enzyme recognition, and optimization strategies. Additionally, we gained relevant materials and software.

Harvest: We processed the harvested information and proposed a new experimental design approach.

4.10 Ailurus Biotechnology

Ailurus Bio. is the world's first bio computer company, focusing on protein synthesis biology solutions. Panda Bio utilizes synthetic biology technology to deeply transform cells into a "micro laboratory" and can simultaneously cultivate billions of cells, achieving large-scale parallel experiments, significantly reducing research and development costs and increasing data yield.

Harvest: Through registration and screening, Ailurus Bio. provided us the PandaPure kit as a sponsor. We had a knockoff meeting with Ailurus Bio. and kept in communication about the progress of the project. They provided guidance on the wet experiment.

5. Public Acceptance

Our concern about the product to the public lies in their views on gene editing technology, so we conducted the survey using the most classic questionnaire method. The purpose of this survey is to understand the respondents' level of awareness, attitudes, and ethical concerns towards CRISPR gene editing technology. The survey questions cover the basic information of the respondents, their level of understanding of CRISPR technology, their views on its potential applications, and their concerns about its ethical and social impacts. By analyzing the survey results, we can understand the public’s perception of our project product. The following survey data was provided by Medicine and Health Care Subject, group 3, Cambridge Summer Programme. DUT-China would like to express its sincere gratitude to them for their external contribution.

According to the data table, the number of valid respondents to the survey is 240. We have selected some key information from the questionnaire for analysis and presented it.

5.1 Age Distribution: The majority of participants are concentrated in the age groups of 18-30 and 31-45, accounting for 50% and 26.67% respectively. Overall, the age of participants is mainly concentrated in the younger group, while the participation of the older group is relatively low.

5.2 Occupational Distribution: The respondent's occupation may affect their level of understanding and perception of CRISPR technology. The analysis results show that students and corporate employees are the two main groups participating in the survey.

5.3 Whether there is a basic understanding of CRISPR gene editing is the basis for making other valuable judgments. There are certain differences in the level of understanding of CRISPR gene editing technology among the survey respondents, with more than half of them not knowing about this technology.

5.4 Further explore the level of understanding among individuals who have claimed aware of CRISPR technology. According to the data table, the majority of people have a certain level of understanding of CRISPR, with the highest proportion having a slight understanding, reaching 69.3%. Overall, most people have some understanding of CRISPR, but the level of understanding is not very deep.

5.5 The attitude of the respondents towards the application of CRISPR in the treatment of genetic diseases is crucial for the public acceptance of our project products. Among them, strong opposition accounts for 1.25%, opposition accounts for 0.83%, neutrality accounts for 48.33%, support accounts for 37.5%, and strong support accounts for 12.08%. It can be seen that the majority of people hold a neutral or supportive attitude, with a higher proportion of support and strong support, accounting for 49.58% of the total number. This indicates that there is significant room for improvement in the public’s impression of gene editing technology. We can gain public trust through improving technological maturity and successful practical cases.

5.6 Ethical issues that respondents believe require special attention when applying CRISPR technology. According to the analysis of the results, the ethical issues that need special attention when using CRISPR technology for gene editing mainly include the safety and long-term effects of gene editing, the ethical boundaries of human gene editing, the ethical issues of editing offspring genes, and the fairness of gene editing.

Among them, the safety and long-term impact of gene editing, as well as the ethical boundaries of human gene editing, are the most concerning issues for respondents. Meanwhile, the ethical issues of editing offspring genes and the fairness of gene editing are second only to the first two. When using CRISPR technology for gene editing applications, the ethical boundaries of human gene editing, the safety and long-term effects of gene editing, the ethical issues of editing offspring genes, and the fairness of gene editing should all be given special attention and response.