HCC has been a long-existing problem in China and it is quite hard to come up with a new method of treatment. Therefore, we must guarantee the reliability of our project in every part of our design. In the past, the oncolytic virus had been widely studied by researchers, so if we want to improve the survival status of hepatic cancer patients, we need to prove our project has been improved in some aspects.
On this page, we show some facts about Proof of Concept based on the project implementation plan and its matched detection hardware. The viability of these parts ensured the reliability of our project.
Our whole project aims to improve the situation of hepatic cancer patients.
Our lab work mainly focused on increasing the efficiency of oncolytic virus. We had confirmed the reliability of the key part for its function. As there are complex interactions between the oncolytic virus and the tumor, a model will prove the reliability of the virulence. Our model had predicted the virulence and the spread of the virus. Besides the treatment part, we noticed that the diagnosis part should be taken into consideration. A new and easy for use method will allow patients to discover the illness at an early stage. Our hardware had developed an easy and reliable way for diagnosis. It had been proved both on the mechanism and the applicability. Moreover, our implementation had proved the marketing possibility of our products. It is a rigorous evaluation on the commercial value of the project, indicating the bright future of our product.
TLR9i special sequence for immunocamouflage
We had managed to cultivate the immune cell in vitro and successfully stimulated its immunoreaction. Under this circumstance, we had proved the immunocamouflage ability of the TLR9i sequence to the immune cell. The result had shown a great reduction of the immune action.
The viability of fusion protein
By fusing the GPC3 scFv with the fiber protein, we got the fusion protein which was able to target the GPC3 receptor. The viability and the characteristic of the protein had been proved in detail. The results are shown on the result page of our wiki.
Effectiveness of scFv
To prove the scFv-fiber fusion protein could specifically target hepatic cancer cells, we had demonstrated the specificity and effectiveness of this targeting by immunofluorescence.
The larger stripe compared with the normal groups indicated the combination between the fusion protein and its target protein GPC3.
The reliability of miRNA modulated suicide switch
We had proved the difference of cytoplasmic free miRNA expression level. The suicide switch in our newly designed E1A gene had shown great regulatory function in normal cells, which had prominently reduced the cytotoxic of the virus to normal cells.
As a newly-developed part, we had made a full evaluation of the structure and the expression ability of this special sequence before we tried to create it in the real life. After we had successfully produced it, we examined whether its ability met our expectations. The result was satisfying. We had witnessed its ability to combine the free miRNA and efficient regulating effect on the suicide switch.
Reduction of targeting genes
Using RNAi mechanism to influence the survival of tumor cells is the killing part of our design. The efficiency of shRNA is the first request for an available design. By transfecting the shRNA-expressed plasmid into HepG2 cells and Hek293T cells, we confirmed their efficiency.
We simulated the key process of the whole progress. This model had taken into consideration of the kinetic mechanism of gene expression, the ligand and receptor combination, and the enzymatic reaction.
The main part of the model is based on ODE functions and the parameters in the model are mostly from published papers. The model makes us have a further understanding of the efficiency of our shRNA killing part, specific proliferation part, etc. Moreover, based on the result of the model, we had also shown the balance between the efficiency of virulence and the cleavage of cells.
As we need to control the number of viruses at the guaranteed lethality notice, this balance is quite important for us. The result of the model had shown the cell proliferation and virus proliferation curves, indicating the validation of our design.To see the full result of our model, please turn to the MODEL page
Our wet lab project aims to develop a clinical-applied drug targeting at reducing the mortality and hazard of HCC. While apart from the actual diagnosis, early detection also counts significantly in tackling the issue of HCC. In order to corporate with our wet lab projects (since our therapy can only be conducted under the circumstance of definite diagnosis), we developed hardware systems aiming at early patient self-inspection and clinical definite detection. To see the full components of our hardware and its result, please turn to the HARDWARE page.
For the efficient and safe use of our products, we had developed a User manual for guiding the right use of the ViruGuard, which concludes most of the points that need to be taken into account. From the production and storage to correct and effective method of application, we have tried to give our guidelines for a how to use it. Besides, we also hope that this user manual can also help us to reconsider whether there are some parts that we haven't considered yet, allowing us to improve and iterate our product in the future.
To have a full understanding to our user manual, please turn to the IMPLEMENTATION page.