Each huge project requires comparative studies and repeated experiments to verify the results from different aspects, also, a clear and innovational idea is essential. Nowadays, the cellular immunotherapy mainly focus on the antigen and antibody, however, tumor heterogeneity makes the target much more diffcult. So we went in the another direction, the microenvironment of tumor is always different from normal tissue, a low-oxygen and acidic environment. Therefore, we wish to develop the TIL cell, a nature targeting cell in the tumor tissue, which is induced by hypoxia inducible promoter to massively secrete our armed- exosome. As tumor cells have a higher efficiency than normal cells in uptaking exosomes, the target consideration is solved to a certain extent.
We’ve already finished the test experiments of high yield of exosome, and miRNA function verification. Following is our future work:
1. The activity of tetracycline induced promoter and galactose promoter was verified using the luciferase reporting system.
2. The anti-tumor effect of miR-942-5p/miR-769-5p was further verified through cell biology experiments, and specific regulatory targets of miRNA were explored using molecular biology experiments.
3. Obtain the approval of our local ethics committee and the informed consent of patients, and then remove tissue and extract TIL cells during the operation of gastric cancer.
4. All modules are constructed on a lentiviral plasmid, and the lentivirus is packaged based on HEK-293T cells.
5. Using lentiviral to transfect TIL cells to construct stable cell lines and animal experiments were conducted with the supervision of the ethics committee. The model of gastric cancer in situ in nude mice was constructed, and the function of our TIL cells was verified by intravenous injection of modified TIL cells.