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√Questionnaire analysis

Through the statistical analysis of the data on the paper questionnaire (Figure 3 and figure 4 are part of the questionnaire data), and with the help of other means, the following conclusions can be drawn:

Rheumatoid arthritis is common in women aged 40-60 years old, so it is a typical middle and old-aged disease, which seriously affect patients' quality of life and family life, so that patients would like not to live any longer. Most patients choose drug treatment to alleviate the pain, but in the actual clinical application, the existing drugs still can not play a very significant therapeutic effect, and that is why patients are eager to the emergence of new treatment methods. They also hope that the new method can have less side effects, and lower recurrence rate, thereby reducing the pain of their own. In addition, rheumatoid arthritis will also increase the burden on the national economy. The United States has ever done a survey that annual rheumatoid arthritis and rheumatoid-related treatment costs more than 1% of its GDP. In China, though there is no exact data, it is estimated that the loss caused by the disease is not less than the United States. And with the aging of the population is becoming increasingly prominent, the national economic burden will become increasingly heavy. If a new treatment can be designed to achieve the effect of radical treatment, this approach will be significant in raising the level of medical treatment and health of its citizenship in countries where population aging is increasing.

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√Finding new treatments

CAR-T, which full name is Chimeric Antigen Receptor T-Cell Immunotherapy, is a chimeric antigen receptor T cell immunotherapy. This is a new cell therapy that has been around for many years but has been improved in recent years to use clinically. Similar to other immunotherapies, its basic principle is to use the patient's own immune cells to specifically remove cancer cells, but the difference is that it is a cell therapy, rather than a drug. As shown in Figure 5, T cells are differentiated and enriched by in vivo injection of cytokines, and then the target cells were separated by cell surface markers. Then, the cells are engineered in vitro and CAR (chimeric antigen Receptor), and amplified by culture to a certain number and then returned to the patient's body, so that the engineering of T cells can be in vivo specific killing of the target cells.

Syn-Notch CAR-Treg cytotherapy

In the daily group meeting (Figure 6), we discussed how to design the treatment of rheumatoid arthritis, and discussed with our advisor in depth. Ultimately, on the basis of traditional CAR-T, we used Treg cells as engineering targets for the purpose of radical rheumatoid arthritis, and we innovatively introduced Syn-Notch system to enhance the stability of engineered Treg cells in vivo. Through the program design and optimization, there existed the final formation of a new Syn-Notch CAR-Treg system (the overall idea shown in Figure 7).

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√Retrospection of Survey

With the deepening of the research on the subject of treatment, the feasibility was verified at the experimental level. But it still had a long way to apply to the clinical treatment and benefit patients indeed. with the results and doubts, we were ready to go to the hospital again, and conducted a questionnaire survey to the patients and interviews to the residents (Figure 8).

Preparatory work:
Respondents: patients and doctors with rheumatoid arthritis at inpatient and outpatient of rheumatology department
Sample size: 30 doctors and 80 patients
Procedure:
1. The production of questionnaire for patients and doctors (Attention to avoid patients’ and doctors’ personal information).
2. The division of staff.
3. Contacting the relevant departments of the hospital.
4. Arranging the route and specific time.

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According to our pre-planned content, we came into the Nanjing First Hospital and Nanjing Gulou Hospital for a questionnaire survey. After lots of conversations, the majority of patients expressed great interest in our innovative therapy and explored the specific clinical use of this therapy. Some patients were confident in our treatment and encouraged us to make persistent efforts to make this therapy really to clinical as soon as possible (Video 1). According to the survey, most of them were sick for more than 5 years and suffered from rheumatoid arthritis pain for a long time. It was gratifying to note that more than half of the patients said they were willing to try Syn-Notch CAR-Treg cell therapy while a part of them showed the uncertainty (Figure 9). But we believe that the recognition degree on this therapy will be higher and higher with the depth of future research.

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In the course of the doctor's research process, most of the them worked for more than 5 years (Figure 10), and they believed that the existing drug treatment had shortcomings like high side effects and recurrence rate and long-term medication (Figure 11), while Syn- Notch CAR Treg cell therapy had a clear target and less toxic side effects (Figure 12), so Syn-Notch CAR-Treg cell therapy has a great advantage over existing treatment, and its safety in the professional also be further confirmed. But its application on clinical treatment needed to overcome many difficulties, and to meet more challenges, such as large-scale production process restrictions, treatment methods remained to be studied, and production quality control standards sitll nedded to be improved, etc.

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