Team:NMU China/Excellence in Another Area

Combatting COVID-19
Naval Medical University CHINA


       Elimination of the SARS-CoV-2 and balance the immune response are both important for late-stage COVID-19, however, no ideal therapeutics can achieve the two goals for our knowledge. To address this, we have
               1) designed synthetic antiviral macrophages with immune-regulatory capacity
               2) demonstrated its feasibility through both in vitro experiments and mathematical modeling.
       The designed synthetic antiviral macrophages, termed "Toggle Macrophages", have the phagocytic capacity specific for SARS-CoV-2 and showed pro-inflammatory/anti-inflammatory phenotypic transformation according to the level of local IL-6 concentration. Therefore, as "smart" therapeutics, Toggle Macrophages can both reduce the viral load through phagocytosis and balance the immune system via phenotypic transformation. We further mathematically modeled and analyzed the feasibility of the practical application of Toggle Macrophages in humans. This work supports the use of Toggle Macrophages as a novel Genetically Engineered Cellular Platform that may be useful in future therapeutic approaches against severe COVID-19.

Background analysis

       The ravages of the SARS-CoV-2 have become a major public health event in the world, which has caused an immeasurable negative impact on the world's economic growth and people's physical and mental health. As medical students, we have been thinking about how we can deal with this problem and hope to work together with researchers from all over the world to overcome COVID-19.
       Academic researchers, pharmaceutical companies, and government agencies have focused their joint efforts on vaccines and antiviral therapies, but a large number of studies have shown that immune pathological damage, especially cytokine storms characterized by high levels of peripheral cytokines, may be the culprit responsible for the deaths of people infected with SARS-CoV-2. On January 24, 2020, the first clinical data report of SARS-CoV-2 infection cases published by The Lancet pointed out that cytokine storms can be seen in critically ill patients. Recently, Academician Zhou Qi of the Chinese Academy of Sciences also stated that the cytokine storm is an important node in the transition of patients with new coronary pneumonia from mild to severe and critical illness, and it is also a cause of death from severe and critical illness.
       After the virus invades the body, the infected cells will release inflammatory factors. These inflammatory factors can dilate blood vessels. Vascular endothelial cells express IL-8 and other cytokines. Immune molecules (such as complement) in the plasma enter the tissues, while cytokines can It sticks the migrating neutrophils and makes them pass through the blood vessel wall into the infected tissues, killing pathogens, and recruiting macrophages. When neutrophils are unable to destroy the pathogen, they will release cytokines again to recruit macrophages to reach the site of infection to phagocytose and kill the pathogen. After phagocytosing pathogens, macrophages will release IL-6, IL-1 and other cytokines, which will stimulate the bone marrow to release a large number of neutrophils to the battlefield, further expanding the inflammatory response; will stimulate liver cells to synthesize a large number of acute phase proteins to participate in complement The lectin pathway works, and the C3a and C5a produced have inflammatory and chemotactic effects, and expand the inflammatory response; at the same time, it also stimulates the hypothalamic body temperature regulation center to increase body temperature. At this time, the dendritic cells and NK cells in the tissue are activated by cytokines, and the dendritic cells take up the antigen to find the initial T cells. Because the pathogen enters the blood, the initial B cells also recognize the antigen. NK cells start to kill pathogens...
       If the immune system defeats the virus, the patient will soon heal. Otherwise, the immune system at this time is like an entire death squad. The enemy is already in the city, facing an extremely fierce attack, his own death is unavoidable. In order to avoid causing more damage to the body, the immune system is now ready to go and convene. The reserve force of the whole body releases various cytokines and makes every effort to fight the virus last. In order to reduce the virus content in the body, I would rather sacrifice myself. As a result, the cytokine storm begins, and finally causes a systemic inflammatory response in the body, which can cause a variety of tissue damage, a variety of organ failures and even death. Therefore, to some extent, the harm caused by the cytokine storm even exceeds the harm caused by the virus itself.
       Therefore, the development of effective treatments for the cytokine storm in the context of the new crown infection has become urgent. In order to avoid the deterioration of the new crown epidemic, there is currently no specific clinical treatment for infection-induced cytokine storms, and non-specific combined treatment measures such as anti-infective drugs, glucocorticoids, nutritional support, and artificial ventilation assistance are often used. Use hormones to suppress the immune response so that the immune response is not too strong. According to the principle that artificial liver can remove inflammatory factors in patients with liver failure, the team of Academician Li Lanjuan previously used this technology for the treatment of severe H7N9, reducing the mortality rate. The “Diagnosis and Treatment Plan for Pneumonia Infected by the New Coronavirus (Trial Version 5)” issued by the National Health and Construction Commission recommends, “Under conditions, for critically ill patients with high inflammation, extracorporeal blood purification technology can be considered.” Renal replacement therapy (CRRT): It is a new blood purification method, which is used more in intensive care unit (ICU). This method can effectively remove inflammatory cytokines in critically ill patients.
       However, these treatments are all treatments after the occurrence of cytokine storm. Is it possible to prevent it from happening, to "cure the disease before it," and to intervene effectively before the cytokine storm occurs? This is the goal of our project.

Policy analysis

       On February 9, 2021, the National Health Commission released the "Reply to Recommendation No. 4371 of the Third Session of the Thirteenth National People's Congress" on its official website: The Commission has always encouraged and supported the research, transformation and industrial development of stem cells and immune cells. Cell preparations such as stem cells and immune cells have obvious pharmaceutical properties. The national drug regulatory authority has formulated supporting policies for relevant preparations to pass drug approval, which can be quickly and widely used after approval, which is not only conducive to ensuring medical quality and safety, but also conducive to industrialization and high-quality development.
       On May 10, 2001, the general office of the State Council issued the implementation opinions on Comprehensively Strengthening the capacity-building of drug supervision (GBF [2021] No. 16). The document points out that the scientific research on drug supervision will be incorporated into the relevant national science and technology plans, focusing on supporting the scientific research on Supervision in the fields of traditional Chinese medicine, biological products (vaccines), genetic drugs, cellular drugs, artificial intelligence medical devices, new materials for medical devices and new raw materials for cosmetics, and accelerating the R & D and listing of new products. At the same time, China's Guangdong, Liaoning, Zhejiang, Tianjin, Hainan, Qingdao, Nanjing, Kunming, Shijiazhuang, Shanghai, Shenzhen, Hangzhou and other places have issued policies to encourage immune cell therapy research and relevant guidelines.
       It can be seen that China's national policy greatly encourages the research on immune cell therapy, which is conducive to the implementation of our project.

Market analysis

        Although CAR-M therapy has not officially entered the market, CAR-T therapy similar to it has shown gratifying curative effects. At present, dozens of companies in China have deployed CAR-T therapies, and many have entered the clinic, including WuXi Juno, Legend Bio, Keji Pharmaceutical, Yimiao Shenzhou, Sibiman, Reindeer Medical, Heng Rundasheng, Genxi Biology, Huadao Biology, Stansai, Ucardi, Jikai Gene, Shanghai Cell Therapy, Boshengji, etc. In particular, CAR-T therapy has achieved great success in the treatment of hematological malignancies. Up to now, the United States has approved 5 CAR-T therapies, and China finally ushered in the first CAR-T therapy in June this year—Akirensai injection. On September 3 this year, the State Food and Drug Administration announced that WuXi's CAR-T product targeting CD19, relma-cel (relma-cel, trade name: Benoda), has officially won Batch. The indications approved for Ruiji Orenza injection this time are: for the treatment of relapsed or refractory large B-cell lymphoma (r/r LBCL) in adult patients after second-line or above systemic treatment. Ruiji Orense Injection is the second CAR-T product approved in China and the first CAR-T product of China's first category of biological products. Akirensai injection and Ruijiolunsai injection have been approved for marketing. First of all, a new treatment option has been added to patients who meet their indications, and there is also a hope of cure. Secondly, this approval means that CAR-T therapy has been accepted by the domestic environment, and the development and approval of more CAR-T products or indications will be accelerated in the future. CAR-T therapy was also successful in my country for the first time in September this year. 28 days after the patient received this therapy, the cancer cells in the human body were successfully eliminated and the cure was achieved. This is the first successful case of CAR-T in China, which is very exciting.
        In order to understand the public's awareness and intention of cell therapy, we put out a questionnaire to 300 people. The content covers the understanding, acceptance and reasons of cell therapy, etc. The questionnaire shows that in terms of acceptance, 91% of the population are willing to receive cell therapy treatment if necessary, expecting it to be safe, effective and economical. Combining the performance of cellular immunotherapy in tumor treatment in recent years, it can be seen that cell therapy has a wide range of applications in the future. Among the 9% of people who are unwilling to receive cell therapy, it is mainly because of ignorance and safety issues, followed by economics. It shows that people's tendency to accept the treatment mainly depends on whether they trust the treatment. This shows that this type of therapy has a huge market space.(more details in Education

        SWOT analysis is based on the situation analysis under the internal and external competitive environment and competitive conditions. We list the main internal strengths, weaknesses, external opportunities and threats closely related to the research object through investigation and arrange them in the form of matrix. Then use the idea of system analysis to match and analyze various factors, so as to make a comprehensive, systematic and accurate research on the situation of the project. According to the research results, we formulate corresponding development strategies, plans and countermeasures.

Risk analysis

       (1) We are concerned about the company's subsequent profitability. Competitive pricing for Toggle Macrophages can be implemented to embrace the different economical background of our customers (COVID-19 patients or patients with cytokine storm). Toggle Macrophages are expected to be covered by every national health insurance program and private insurance companies.
       (2) Security is a huge risk. We try to avoid some security problems and make sure that no personal or environmental harm occurs.
       (3) We are not yet graduated and we lack experience and certain capacities. Furthermore, our project is in the very early stages of development, so there are still many phases to go through before getting to the destination. All this research and development is a high-risk investment. But a full understanding of consumers’ needs and comprehensive market analysis have been carried out before the initiation of Toggle Macrophages.
       (4) Finances are essential. If we want to start up, we will have to organize ourselves in a way different from what we do in iGEM project. We have managed to raise funds for the company. We will continue to work hard in the future.

Product design and development plan

       Our project has many innovations and highlights.
               (1) Engineer a new type of macrophages, MS, which can both engulf viruses and release no cytokine.
              (2) A novel cytokine receptor, the IL-6 receptor, was designed to be able to make different responses depending on the level of cytokines (IL-6) in the environment.
               (3) Offer a whole new solution for controlling the cytokine storm caused by the virus.
               (4) It can be generalized to other diseases that cause cytokine storm.
       To bring these highlights to fruition, we did the following planning for the course of the project, with specific timing to continue adjusting based on subsequent progress. Now, we are at the first phase and we have the opportunity to continue our project after the iGEM competition.

Team organization

        Enterprise organizational structure is the most basic structural basis for enterprise process operation, department setting and function planning. We arrange the organizational structure of the company according to the characteristics of the project to achieve the reasonable operation state of "1 + 1 > 2".

Financial analysis

        We used the following further assumptions: in the first few years, the company can be financed with angel investment and venture capital. After the first few years, the company’s costs can be covered by its own revenues, meaning that we reach break-even and start to make profits. After the 10th year, the company’s revenues are expected to grow by 5% each year.
        Guosen Securities is a large national comprehensive securities company in China. By the end of June 2021, its registered capital was 9.612 billion yuan, with outstanding innovation ability, competitive advantage and market position. On October 13, we came to Guosen Securities, introduced our project to them, communicated with them and reached a preliminary intention. After the iGEM competition, we will continue to follow up the project.

       We also took into account the main expected expenses of our company. We based our assumptions of costs either on different reference material or expert talks.