Through the Molecular Docking studies, the research team of the Oncotherapy Project of the Ebris Foundation designed a library of potentially active molecules on a protein belonging to the family of MDR (Multi Drug Resistance proteins) known to be expressed in numerous resistant tumor forms. They therefore selected and synthesized, using our chemical and pharmaceutical skills, a series of molecules on which we performed proteomics experiments (DARTs) to confirm the selected target, choosing some cell lines resistant to chemotherapy such as mitoxantrone (including HepG2, MCF7-transfected and HT29). Subsequently, in-cell assays were performed to test the inhibitory potency of our compounds by evaluating their cytotoxicity in co-administration with the reference chemotherapeutic agents.
The most promising molecules will subsequently be tested in a human colon organoid system. Starting from surplus material resulting from surgical removal of colon cancer in eight naive cancer patients, eight tumor samples and the same number of healthy mucosa were obtained, provided by the National Cancer Institute "G. Pascale Foundation"
In particular, a protocol for the preparation of 3D cell cultures in micro-fluidics was developed in the laboratories of the Ebris Foundation, which induces the production of extracellular matrix (ECM) resulting from an active interaction between tumor cells and stromal cells, mimicking what happens in vivo.
In addition to the realization of cell cultures in 3D, the collected tissues were fragmented and stored in cryogenic tubes in order to set up a bio-bank of tissues to be used both in view of the application of protocols to create organoid cultures from frozen tissues and through flash frozen technique, for genomic and proteomic sequencing. On these organoids obtained, the goal is to confirm the validity of the spheroid system to monitor an antitumor effect, by administering 5FU as a reference chemotherapy for the treatment of colon cancer. Tumor growth will be monitored and cellular system death due to drug action will be observed. Subsequently, the expression level of the protein will be evaluated by qRT-PCR. Once the expression of the aforementioned proteins in our organoid system has been confirmed, it will be possible to co-administer the synthesized inhibitors with the chemotherapy to assess whether the drug better induces tumor death. It will be possible to verify a potential increase in cytotoxicity of chemotherapeutic agents such as Mitoxantrone, whose biological activity is often limited by the presence of ABC proteins.