Translating from the laboratory to the real world requires cautious planning at every step. From the perspective of safety and target audience, our project aims to deliver our products to the end users in the safest form. Through communication with researchers in cancer research, we worked out a practical implementation plan for the project and hoped to provide practical and safe help to cancer patients.
Cancer poses a terrible threat to human health, and millions of people worldwide die of cancer every year. The RSPOs secreted protein family is the main cause of many cancers. At the same time, traditional therapies are limited to many issues such as toxicity and side effects. Based on the above two points, NEU_CHINA developed RTAC and took engineered S.Cerevisiae as vehicle to deliver RTAC to solve the problems, hoping to provide opportunities for future cancer targeted treatment.
Patient:
We designed RTAC for patients to antagonize multiple RSPO hyperactivation-caused cancers, including colorectal, breast or lung cancers. And as a probiotic, R-yeast was specifically designed for colorectal cancer patients. Through the research of integrated human practice and education, we learned that many patients are suffering from cancer symptoms and existing treatments. We earnestly hope that cancer patients can benefit from our project and improve their quality of life.
Scientific researcher:
Our experimental ideas can provide some inspiration for researchers studying cancer targeted therapy. The creation of RTAC can provide researchers with a new scheme for targeting drug design, and meanwhile R-yeast further expanded the notion and usage of probiotic.
Healthy people:
Though not fully interrogated, the notion that R-yeast can be used as probiotic for colon cancer prevention is appealing. RSPO-hyperactivated colon cancer came from hyperplasia and adenoma, in which RSPO functions to promote the tumor progression. However, early formation of hyperplasia and adenoma is not easy to detect or diagnose. Since R-yeast is sensitive to the eATP at the tumor tissue, it can serve as a preventive probiotic to neutralize the tumor formation from the beginning. Also because of the tumor-specific sensing machinery, R-yeast may exist as a commensal S.Cerevisiae in the healthy intestinal tract without creating health issue. Further test of this theory on lab animals should be conducted in the future.
After we successfully designed RTAC and used yeast to carry drugs, we began to verify its effects in many ways, hoping to make our project move from the laboratory to the real world.
RTAC itself has good flexibility and solubility, strong stability, and adequate secretion capacity.
RTAC exhibited pan-RSPO binding capacity towards all RSPOs, which was stronger than LGR4 or ZNRF3 ECD alone.
RTAC blocks in vitro tumor cell proliferation and RSPO-mediated Wnt signaling, and has been proved broad effectiveness in a variety of tumor tissues.
See more in “Results”
The expression of RTAC in R-yeast remains silent in the absence of eATP and will auto-sense and switch on when coming across tumor tissue, which enhanced targeting of RTAC.
R-yeast significantly inhibited the Wnt-target genes expression and colon cancer cell growth in response of eATP.
See more in “Results” for details
Experiments will be conducted at lab animal level to verify the effect of RTAC and R-yeast in vivo.
Select the appropriate medication method: we plan to use embedding technology to make tablets and take probiotics orally.
We have collaborated with LZU-CHINA to do a preliminary attempt on using AC (Alginate - chitosan) microencapsulation to cover our probiotics. At first, our team did background analyzing of the usually used methods to cover probiotics and finally decided to use AC microencapsulation because of the advantages it has. Then after finding out how could this method work, we also found evidence of the feasibility of using this method to achieve our goals by researching previous studies.
The background analyzing document done by our team is shown as below.
Then, LZU-CHINA found literatures and protocols to do experiments, the result was shown as below.
Carry out clinical trials in the near future to achieve real connection with patients.
In order to ensure that probiotics will not harm the human body after entering the intestinal tract, we have selected Saccharomyces Cerevisiae, which is most commonly used in food production, as the carrier to prevent adverse effects on the intestinal microenvironment.
We developed a novel kill switch to prevent our engineered yeast from accidentally leaking and affecting the normal life activities of other cells or organisms. This design allows the engineered yeast to survive and grow only in human intestine where copper ion is rich, or culture medium with copper ion addition, while leading to the death of the engineered yeast in normal environment.
Since RSPO is required for human tissue homeostasis, we implanted the self-tunable switch in R-yeast to allow the specific expression of RTAC only in response to tumor microenvironment. In the sense, the level of RSPO can be finely tuned-down to stop the cancer progression at the tumor tissue locally while posing low threat to normal tissue regeneration and repair.
See more in “Safety”
From R&D to marketing, drugs need to go through the process of pharmacological research - toxicological research - preparation development - clinical trial - DNA application data - clinical monitoring period - approval, which lasts for a long period and requires the unremitting efforts of researchers.
Our project adopts Saccharomyces cerevisiae as the carrier, so it is necessary to make the engineering fungi colonize in the intestinal tract. The number should remain stable within a certain range, neither too much can affect the intestinal microecology balance, nor too little can inhibit the tumor. This can only be explored through individual level experiments.
Our project takes colorectal cancer with overexpression of RSPO protein as an example to study. Our goal is to achieve multiple cancer treatments targeting high activation of RSPOs. Therefore, for different cancers, we are faced with the problem of finding specific characteristic molecules, drug carriers and transportation modes according to the characteristics of different cancer environments.