Biological brachytherapy with the use of transduced adipose tissue stem cells (ADSC)

Scientific goal: The overall goal of the project is to create a rat model of modified adipose tissue stem cells producing the TRAIL protein with anti-tumor activity, administered to free tissue lobes. Another goal is to develop a complete animal model of biological brachytherapy. State of knowledge: Tumors occurring in the superficial layers of the human body (skin, breast, head and neck area) require, after radical resection, adequate restorative therapy. To date, no easy-to-implement and safe model for creating functional tissue flaps that, in addition to restoring the appearance of the tissue, affects the root cause of the disease, has not been developed. There are few reports on biological brachytherapy - the process of creating modified tissue flaps, exerting an additional, local therapeutic effect on the primary conversion, e.g. anti-cancer, accelerating the healing of chronic wounds, angiogenesis. Until now, genetically engineered adipose tissue stem cells (ADSC) have not been used to develop such tissue flaps. The study is translational, and if the effectiveness of functional tissue flaps is proven, the collected data can be used to implement an analogous solution in breast reconstruction after mastectomy due to breast cancer. Material and methods: The project consists of successive tasks, which, performed sequentially, will allow for the development of a complete rat model of biological brachytherapy. First, the surgical procedure will be optimized - the lifting of the subcutaneous flap from the superficial inferior epigastric vessels and the perfusion of the flap. Subsequently, ADSC cells will be isolated from rat adipose tissue, followed by lentiviral transduction with fluorescent protein vectors. The modified cells will be used to optimize the perfusion of the tissue flaps. In addition, ADSCs will be transduced with the TRAIL protein with lentivirus vectors. The cytotoxic effect of such modified cells on the rat breast cancer cells RBA will be investigated in vitro. Then, the TRAIL-producing ADSCs will be applied to the free skin flap under which the RBA breast cancer tumor will be induced. The effectiveness of administering the modified cells to the flap will be confirmed by: observation of the animals' survival, vivid visualization of the dynamics of tumor growth and microscopy (post mortem). Expected end result: The result of the study will be a complete verification of the effectiveness of modification of free tissue flaps by enriching them with transduced mesenchymal stem cells from adipose tissue. In addition to objectively assessing efficacy, the study will accurately describe the model of rat biological brachytherapy with autologous modified cells. Hence, it will be possible to use the results of the study for comparative studies of other tumors/anti-cancer proteins.